Despite the growing interest and rapid advancement of alternative photovoltaic (PV) technologies such as perovskite based PV devices, we still believe that organic photovoltaic (OPV) devices have a significant potential for stable, low-cost solar power generation. As a matter of fact, Heliatek announced a new organic photovoltaic world record efficiency of 13.2% in the beginning of February 2016, highlighting the continued interest in OPV technology from both academia and industry [The Special Issue on organic photovoltaics that follows highlights the complexity of the issues facing researchers and engineers involved in OPV research and development. Interestingly, the author contributions cover the field from macromolecular engineering to device elaboration and characterization, not forgetting active layer morphological control, characterization of bulk heterojunction (BHJ) solar cells and bilayer device architectures.et al. present a novel approach based on the use of spirocyclic systems in the conjugated backbone of electron donor polymers to tune the energy levels of the materials [et al. review extensively the various and sometimes contradicting effects of backbone fluorination of conjugated polymers in use for solar cells [et al. point out the importance of singlet exciton lifetime in electron donor polymers in regards to device performances, i.e., photocurrent generation [In terms of molecular and macromolecular engineering, two interesting contributions discuss new methods to tune the optoelectronic properties of the electron donor polymer, while a third contribution is proposing an analysis of the effect of molecular architecture on photocurrent generation. First, Bronstein aterials . Modificar cells . The disneration . The autet al. propose to (re)consider bilayer architecture with an optimized sequential solution deposition technique, allowing the control of the morphology at the single interface between the electron donor polymer and the acceptor molecular system [Regarding the active layer morphology, donor–acceptor phase separation control is still a key issue in the OPV area. To face this problem, Jang r system . The proet al. have also been working on the issue of the donor–acceptor phase separation in BHJ systems, but from a characterization point of view [Dyck  of view . They deet al. point out the importance of organic solar cells lifetime problems and the lack of outdoors tests and data [Finally, Owens and data . The autTo conclude, in order to reach both high efficiency and long-term stability of organic solar cells, only a well-balanced optimization of all the different criteria  will be effective. For this reason, it is important to encourage interdisciplinarity and interlaboratory collaboration approaches to face all the remaining issues preventing the pre-industrialization step of the OPV area.


Currently, immunotherapy is considered to be one of the effective treatment modalities for cancer. All the developments and discoveries in this field up to the recent Nobel Prize add to the interest for research into this vast area of study. Targeting tumor environment as well as the immune system is a suitable strategy to be applied for cancer treatment. Usage of nanoparticle systems for delivery of immunotherapeutic agents to the body being widely studied and found to be a promising area of research to be considered and investigated further. Nanoparticles for immunotherapy would be one of the effective treatment options for cancer therapy in the future due to their high specificity, efficacy, ability to diagnose, imaging, and therapeutic effect. Among the many nanoparticle systems, polylactic-co-glycolic acid (PLGA) nanoparticles, liposomes, micelles, gold nanoparticles, iron oxide, dendrimers, and artificial exosomes are widely used for immunotherapy of cancer. Moreover, the combination therapy found to be the more effective way of treating the tumor. Here, we review the current trends in nanoparticle therapy and efficiency of these nanosystems in delivering antigens, adjuvants, therapeutic drugs, and other immunotherapeutic agents. This review summarizes the currently available bioactive nanoparticle systems for cancer immunotherapy. The treatment for cancer has been studied and explored for decades. Advancements and breakthroughs in this area have changed the outlook of cancer treatment options. A number of therapeutic options are currently available for treatment, and many are under clinical trials. Cancer can be treated utilizing many methods, such as surgery, chemotherapy, radiotherapy, hormone therapy, and immunotherapy. Among these modalities, surgery has been used broadly as a treatment modality. However, the reoccurrence of tumors is the main drawback of this treatment option. The side effects of chemotherapy, hormone therapy, and radiotherapy are also considered to be a downside of the current cancer treatment. These kinds of drawbacks in treating the tumor have led to approaches for more specific targeting of the tumor site with fewer side effects ,2.Among the number of treatment options for cancer, immunotherapy is one of the effective methods due to its specificity in boosting our natural defense mechanism against cancer. The important milestone in the evolution of cancer immunotherapy is considered to be the manipulation of the immune system at the molecular and cellular levels ,4,5. AftBoth off-target side effects, cost, and lengthy processing are considered to be drawbacks of immunotherapy treatment methods ,11. The The physicochemical characterization is also an important factor to be considered. The size, charge, surface modifications, stability, toxicity is closely related to the action of these particles in the body. The size, shape, charge, and capacity to incorporate therapeutic agents inside the nanocarrier make the nanoparticle system suitable for therapy. Currently, liposomes, PLGA (polylactic-co-glycolic acid) nanoparticles, dendrimers, gold nanoparticles, micelles, polymeric nanoparticles, and microneedles are widely studied for targeting particular immune cells or immune reactions. The combination of biomaterials and drugs using nanotechnology has resulted in interesting outcomes regarding the treatment strategies ,16,17,18Our health is maintained by the immune response as one of the key factors. The tumor immune response is complex; however, many studies have been conducted to discover the interesting mechanisms behind the response. As is known about innate and adaptive immunity, both of these immune responses together help the body maintain its equilibrium ,25,26. DBoth the innate and adaptive immune responses are different and complex in nature. The innate immune system mainly acts through the phagocytosis pathway, which is composed of phagocytic cells such as natural killer (NK) cells, macrophages, neutrophils, and monocytes. The main mode of action of the innate immune response is to discriminate the proteins presented by these cells from those of invading cells. The factors that are able to discriminate these presented proteins from those of the tumor cells are called major histocompatibility complex class I (MHC-I) ,32,33,34Immune checkpoint blockade as a therapeutic modality that has gained more attention after the modification of CAR T cell-modified T cells to block specific immune regulatory checkpoints. The advancement in checkpoint blockade therapy resulted in the invention of the PD-1 (programmed cell death-1) and PDL-1 (programmed cell death ligand-1) blockade method, which is characterized by both the specificity and the improved outcome. Another intracellular protein present in T cells, cytotoxic T lymphocyte-associated protein-4, can also be utilized to inhibit the regulation of T cells followed by the arrest of both the proliferation and activation of the tumor cells ,44,45,46The mode of action of the immune response against tumors is based on how the immune system can differentiate between cancer cells and normal cells. The main difference between cancer cells and normal cells is the expression of various kinds of TAAs in cancer cells. The killing of cancer cells can be achieved by both immunogenic and tolerogenic signals. The immunogenic signals are released due to the rapid growth of the tumor followed by necrosis. The necrosis of the tumor will result in the release of TAAs and immunogenic signals followed by the activation of phagocytes and antigen-presenting cells (APCs). This kind of activation results in the production of cytokines such as interferon gamma (IFN-γ) and tumor necrosis factors, leading to the upregulation of MHC class I expression and tumor suppression. The tolerogenic signals can act in main three ways. The downregulation of MHC class I and killing of tumors by TAA-specific cytotoxic T lymphocytes (CTLs) is considered to be one way of killing tumors. In another way, CTL-mediated killing mechanisms can evade immune surveillance. Finally, tumor killing occurs by programmed cell death PD-L1 via activated T cells .A number of cancer immunotherapeutic approaches have been explored widely. However, many new studies have utilized dendritic cell (DC) vaccines, immune checkpoint inhibitors, monoclonal antibodies, cytokines, and adoptive cell therapies . The modThe efficacy and safety of this immunotherapy opened a new window for clinicians to explore and evolve for the betterment of cancer therapy. The discovery of immune checkpoint blockade and CAR T cells for inhibiting tumor growth led immunotherapy to be the suitable modality for cancer treatment. Newer monoclonal antibodies, targeting new immune system checkpoints, the invention of tremendous cancer vaccines and oncolytic viruses, the modification of CAR T cells and the discovery of tumor-infiltrating lymphocytes and interleukin-2 make this area of treatment different from other treatment methods ,52,53.Bioactive nanoparticles are considered to be the prime option for immunotherapy due to their special characterizations, such as size, shape, charge, elasticity, and ability to work as a carrier ,55. NanoAnother major factor that affects the internalization of nanoparticles into cells is the charge of the nanoparticles. The surface modification of nanoparticles can be accomplished by applying various surface chemistries. Cationic particles accumulate more in tumor cells. Modification of a nanoparticle surface to obtain a positive charge can be utilized. Hence, cationic particles show higher toxicity, and advancements in modification are currently being investigated by many researchers. These three modifications are the major ones to be considered before synthesis of a nanocarrier or nanoparticle for immunotherapeutic use. Other than these modifications, the nanoparticle ligand density and nanoparticle elasticity can also be tuned for improved transport and accumulation of the nanoparticles  58]. Th. Th58]. The various bioactive nanoparticles enable them to be the primary option for immunotherapy . The appPLGA is considered to be an excellent component for immunotherapy due to its low systemic toxicity and high biodegradability compared to other polymeric systems. The majority of PLGA-based nanoparticle studies for cancer immunotherapy are based on targeting the nanoparticle system to DCs. PLGA nanoparticles are taken up by DCs without any recognition of this specific character ,66. TargThe antitumor cytotoxic T cell response can be induced by the delivery of vaccines to the DC by targeting DCs using PLGA nanoparticles. CD40-mediated delivery of vaccines to DC cells using PLGA nanoparticles resulted in an antitumor response with prolonged survival. This study reveals that PLGA nanoparticles could be a perfect option for targeting DCs to deliver vaccines . PLGA naThe combination of PD-1 blockade and laser immunotherapy can also be achieved by PLGA nanoparticles. After the intratumoral injection of gold nanoshells and anti PD-1 peptide (APP)-loaded PLGA nanoparticles, an excellent killing effect at the primary tumor site was achieved by photothermal therapy (PTT). These nanoparticles also played a vaccine-like role and induced a localized antitumor-immune response. APP release with PTT transient triggering could induce the blockage of the PD-1/PD-L1 pathway to activate T cells, thus subsequently generating a systemic immune response. This therapeutic platform could efficiently kill primary cancers and treat the growth of metastatic tumors .Liposomes for the delivery of cancer immunotherapeutic agents are widely studied among different lipid-based nanoparticles. Co-delivery of ovalbumin (OVA) and IFN-γ-encoding pDNA to the DCs by utilizing pH-sensitive liposomes was found to be an effective treatment option. The combined therapeutic effect of OVA and IFN-γ-encoding pDNA in tumor-bearing mice promoted the infiltration of CTLs into the tumor and resulted in a strong antitumor effect . AntigenRecently, Yuba E et al. studied the strong immunotherapeutic effect of pH-sensitive dextran liposomes modified with PEG as well as TGF-β 1 (transforming growth factor) receptors. The liposome delivery resulted in a high antitumor effect by the infiltration of CD8-positive cells to the tumor . Yuba etTo overcome the immunosuppressive tumor microenvironment (TME), it is very important to enhance the efficacy of cancer immunotherapy. Agonists of stimulator of interferon genes (STING), a cytosolic immune adaptor protein, have been shown to induce potent antitumor activity when delivered into the TME. The anionic properties of STING agonists make them poorly membrane permeable, cationic liposomes with varying surface polyethylene glycol levels, and they can be used to encapsulate 2′3′-cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) to facilitate its cytosolic delivery. Liposomal delivery improves STING agonist activity and results in immunological memory that helps rechallenge tumor cells .Another area of immunotherapy by gene delivery also utilizes liposomes as the carrier for specific targeting. The delivery and targeting of RNA to DC cells using lipoplexes also has gained more attention in cancer immunotherapy. The delivery of RNA lipoplex (RNA-LPX) to DC cells activates INF-α-mediated immune mechanisms, resulting in in situ DC maturation followed by innate immune reactions. The results were promising for cancer immunotherapy; therefore, RNA-PLX systems can be utilized as a better therapeutic agent for DC targeting .Generally, PEGylation is commonly used in these kinds of studies to modify lipid nanoparticles for the delivery of siRNA. A pH-sensitive cationic lipid called YSK05 for the formulation of a multifunctional envelope-type nanodevice (MEND) is used as a carrier for siRNA to overcome the limitations followed by optimization of the lipid composition. The intratumoral administration of the PEGylated YSK05-MEND enhanced gene silencing and was found to be a good agent for delivery of siRNA into the cytosol .The use of micelles for cancer therapy has been studied and explored both preclinically and clinically. There is a wide range of applications of micelles in cancer treatment as carriers for imaging, chemotherapy, radiotherapy, and immunotherapy. The synthesis of micelles is comparatively easier than that of other nanoparticles. The biodegradability and nontoxicity of these formulations make them suitable for carrying therapeutic payloads . AntigenAnother study showed the stimulation of the immune response using micelles with the combined action of PTT with immunotherapy. The immune response can be stimulated by regulating metabolism-related enzymes. Due to the accumulation of IR780 in the tumor followed by migration to the lymph node, PTT can be performed, resulting in the inhibition of IDO . The inhibition of IDO leads to the activation of T lymphocytes followed by the inhibition of distal tumor growth . Combined therapy of this micellar system kills the tumor by PTT and inhibits distal tumor growth post-PTT in vivo in BALB/c-nu mice .Polymeric micelles are currently considered to be a more exploratory nanoparticle carrier system for cancer immunotherapy. The delivery of tyrosinase-related protein 2 peptide antigen and adjuvant to the lymph node in the B16F0 melanoma mouse model was performed using cationic diblock polymeric micelles. The resulting increased T lymphocyte anticancer activity indicates the efficiency of these kinds of micelles in treating cancer by improving the immune response .Repolarization of Tumor associated macrophages (TAMs) to M1 macrophages can be used as a strategy for cancer immunotherapy. The use of galactose-functionalized zinc protoporphyrin IX grafted polypeptide micelles for targeting TAMs and delivering immunopotentiators resulted in the induction of ROS and decreased STAT3 expression. Activation of T lymphocytes by the repolarization of TAMs resulted in tumor regression .Gold nanoparticles for immunotherapy are considered to be a highly promising area of research due to their favorable characteristics. Recently, a number of studies have indicated that the ability of gold nanoparticles makes them a perfect carrier for immunotherapy . These nDelivery of antigens using different gold nanoparticles is possible due to their high affinity towards antigens; at the same time, the affinity depends on the size and shape of the gold nanoparticles that are used. Delivery of CpG for immunotherapy was attempted in different sized and shaped gold nanoparticles, such as gold nanorods, nanoshells, and nanostars of different sizes. The size of the nanoparticle also plays a very important role in the delivery of antigens to the desired site. It has been observed that the 50- and 15-nm gold nanospheres are the perfect option for immunotherapeutic delivery of antigens .Delivery of antigens/adjuvants such as OVA and CpG using gold nanoparticles for the in vivo B16-OVA tumor model is an effective method to treat cancer by activating the immune system. The results of this study showed an antigen-specific immune response that led to an antitumor response and an improved survival rate . DeliverGold nanoparticles can be used both as potent carriers for therapeutic moieties and as PTT agents. The combined PTT and immunotherapeutic effect of gold is possible due to its optical and therapeutic characteristics. The PTT effect resulted in the expression of cytokines and chemokines followed by DC maturation and the T cell immune response. Gold nanoshells could be used both as a PTT agent and an effective immunotherapeutic agent . Another3O4 as a delivery system of OVA and as an immune potentiator. The delivery of iron oxide nanoparticles alone exhibited both immune cell activation and cytokine production. These results confirm the immunotherapeutic effect of iron oxide nanoparticles by themselves in a colon adenocarcinoma (CT26) tumor animal model [Iron oxide nanoparticles can also be used as potent carriers for vaccine delivery and can be used as antitumor agents for cancer therapy. A very recent study on iron oxide nanoparticles for vaccine delivery showed an improved therapeutic effect. In this study, the authors used a superparamagnetic Feal model . Anotheral model . Other gal model .Dendrimers are used to deliver OVA to immune cells by incorporating OVA into guanidine-terminated dendrimers by utilizing the helix B region of OVA . ImmunodExosomes are basically secreted by immune cells, and they are characterized to deliver proteins and antigens for therapy. The use of biomimetic exosomes for the delivery of cargos for immunotherapy has gained more attention. Delivery of the monoclonal antibody DEC205 to dendritic cells can be performed using biomimetic exosomes. The synthesis of these exosomes is similar to liposomes. The study was a novel method of delivering antigens to DCs for immunotherapy . The synAmong the number of treatment modes for cancer therapy, immunotherapy was found to be more effective with better outcomes. Currently, there are numerous studies being published that combine immunotherapy with other modes of therapy, such as PTT, photodynamic therapies (PDTs), radiotherapy, and chemotherapy. Utilizing immunotherapy with other therapy combinations appears to be a potent way to eradicate cancer and will result in more specific and effective antitumor effects. Overall, combination therapy has resulted in a complete antitumor effect with less risk of tumor recurrence and no metastatic progression. Combination therapy could be a milestone future cancer therapy with low side effects and good results.There are many studies in which PTT and immunotherapy combinations are used to eliminate cancer. The major benefit of combining PTT with immunotherapy using nanoparticles is the relatively lower risk of tumor recurrence . PDT in The combination of immunotherapy with radiotherapy can also be utilized to obtain an improved antitumor effect in different cancer models; hence, the possibility of tumor antigen release is greater after radiation treatment. A plant-based virus-like (VLP) nanoparticle was used for the delivery of radiation therapy in patients with oral melanoma. VLP nanoparticles increased the infiltration of immune cells to the tumor site, followed by an immune response and tumor killing after radiation .Another area of combinational therapy that utilizes chemotherapy and immunotherapy has also gained more attention in cancer therapy. Numerous studies have also been performed based on this strategy to completely eradicate cancer and reduce the risk of recurrence. Immunosuppressive tumor microenvironment (TME)-responsive nanocarriers were used to deliver PTX, mitoxantrone (MIT) and celastrol (CEL). The importance of delivering drugs is to induce both chemotherapeutic and immunotherapeutic effects synergistically. This combined action of drugs resulted in a better antitumor effect and prevented metastatic progression .Immunotherapy is one of the most explored areas in the new era of cancer treatment due to its specificity and fewer side effects compared to the other modality of therapy for cancer. A number of methods have been explored to generate an immune response against cancer by developing new immunotherapeutic agents through the modification of current treatment agents and the development of new carriers for improved targeting. These approaches can be considered a milestone in this area of cancer treatment.A number of different bioactive materials have been studied preclinically and clinically for cancer therapy. The use of biomaterials in the form of nanoparticles also increased the effectiveness of the therapy due to the special characteristics of these nanoparticle systems. The specificity and targeting ability of the nanoparticle can be altered using different therapeutic moieties, polymers, and targeting agents. Liposomes, PLGA nanoparticles, micelles, dendrimers, and inorganic nanoparticles are commonly used bioactive nanoparticle systems for cancer immunotherapy. All of these nanoparticle systems are currently studied widely, and research is ongoing. The advantages of these kind of bioactive systems are their high targeting ability, low toxicity, biodegradability, and high specificity. However cost for cancer immunotherapy also considerably high when it comes to the clinical perspective. The increased cost of nanomedicine limits this therapeutic options to be in mainstreams. However new approaches are coming out to make cost effective nanoparticle systems for future cancer immunotherapy. The major factors which is to be considered in a better clinical outcome would be the efficiency, potency, toxicity, and cost effectiveness of the nanoparticle systems.Delivering antigens/adjuvants or other drugs/vaccines to the desired cells or lymph nodes can be done effectively by the use of these types of nanovehicles and is accomplished more easily than other methods of immunotherapy. Liposomes for cancer treatment are currently being studied at the clinical level and represent a potent carrier system for cancer therapy. There are a number of drugs are being studied clinically for cancer immunotherapy such as atezolizumab, nivolumab, pidilizumab, paclitaxel, etc. The major mechanism of action of these drugs are anti-PD1 as well as anti-PDL1. Some of these drugs are currently in clinical trial III phase and some are in phase I and II. To date, a number of studies have indicated that bioactive nanoparticle-assisted immunotherapy could be a perfect tool for future cancer treatment. All these kind of nanoparticles exhibits some unique characteristics which makes them suitable candidate for immunotherapy. In the case of liposomes, it can load both hydrophilic and hydrophobic moieties inside, which enable the delivery of different kinds of therapeutics into the tumor site. PLGA nanoparticles are food and drug administration (FDA) approved and widely used in clinical field for controlled drug delivery as well as in manufacturing of surgical sutures. At the same time PLGA nanoparticles can be used to target specially to DCs which can be utilized for immunotherapy. In the case of micelles, a number of polymeric moieties can be used to synthesize different types of micelles. These polymers can be responsible for a number of immune response which helps in cancer immunotherapy. Moreover the release of therapeutics from micelles can be facilitated by different stimuli responses which can be utilized in immunotherapy. The advantage of use of gold nanoparticles for immunotherapy is wide ranging from their optical properties to therapeutic effect. It can be used for imaging, deliver of macromolecules for therapy and also for combination therapy such as thermal ablation. All these bioactive nanoparticles are unique and suitable for immunotherapy for cancer. Combined immunotherapy with other treatment methods has also recently gained attention. This combination of PTT, chemotherapy, and radiotherapy with immunotherapy can hopefully contribute to future cancer treatment.


Cynictis penicillata), the rest are from other wild carnivores including the highly endangered wild dog (Lycaon pictus). Lyssavirus infection was confirmed in two wild dogs and a spotted hyaena (Crocuta crocuta) in the Madikwe Game Reserve, North West province in South Africa, in 2014 and 2015, using a direct fluorescent antibody test and immunohistochemistry. There had been no new wild dog introductions to the Madikwe Game Reserve for many years and the wild dogs were last vaccinated against rabies approximately 11 years prior to the incident. The first euthanised wild dog was the last surviving of a break-away pack of 6, and the second was the last of a larger pack of 18, the rest of which died with no carcasses being found or carcasses too decomposed for sampling. Subsequent antigenic typing of the lyssaviruses indicated that they were canid RABVs. The RABVs originating from 22 wild carnivore species, 7 dogs, and a caprine, mostly from the North West province, were genetically characterised by targeting a partial region of the nucleoprotein gene. The nucleotide sequence analyses of these viruses and two previously characterised RABVs confirmed that the outbreak viruses were also canid rabies, phylogenetically clustering with virus isolates originating from black-backed jackals recovered between 2012 and 2015 from the North West province, and domestic dogs from neighbouring communal areas. The source(s) of the mortalities and possible reservoir host(s) for the virus could only be speculated upon from data on specific predator numbers, movements and behaviour, kills, park management and the changing environmental ecology, which were monitored closely in Madikwe over several years. The most likely rabies sources were from boundary fence contacts between wild carnivores within the park, with domestic dogs or cats and/or naturally occurring wild carnivores outside the park. The associated risk of zoonotic infection and threat to important and endangered predators may be mitigated through regional rabies control primarily in domestic dogs and cats, as well as by preventative vaccination of at-risk park employees and their pets. The importance of ongoing prophylactic rabies protection by regular vaccination of highly endangered wildlife carnivores and the submission of carcasses for rabies diagnosis of any wild or domestic animals behaving uncharacteristically or found dead is emphasised.Both domestic and wild carnivore species are commonly diagnosed with rabies virus (RABV) infection in South Africa. Although the majority of confirmed rabies cases in wild carnivore species are reported from the yellow mongoose ( Lycaon pictus), also referred to as the African painted dog, is considered one of the most endangered wildlife carnivore species in sub-Saharan Africa   in the Ethiopian Bale mountains  RABV without repeated introduction of the virus from domestic dogs   were regarded as the principal host species of rabies in the area, it was assumed that the jackal cycle was the source of the infection. Whereas the pups had not been vaccinated, the five adults, all of which survived, had each been vaccinated by the parenteral route at least twice and all had significant rabies serum–neutralising antibodies. This observation showed that multiple vaccinations against rabies were effective in protecting wild dogs against natural challenge, lending support to findings from other studies in captive wild dogs . Through antigenic and genetic characterisation as well as background Madikwe predator and environmental data, the report attempts to identify the possible sources of infection in the park.2, which has been expanded to 750 km2 by newly incorporated private reserves. It was previously degraded farm land and has recovered as a result of sustainable use by a diversity of wildlife including 66 large animal species. The perimeter is 150 km of electrified fencing. The geography is large open grassland plains, woodlands with rocky outcrops and single mountains (http://www.madikwegamereserve.co.za).Madikwe Game Reserve is situated in the North West province of South Africa and is bordered on the west by Botswana and was announced in 1991. It is spread over 680 km®  vaccination as prescribed by the manufacturer. A month later, a second wild dog, the last surviving member of a larger pack of 18 wild dogs, which had also presumably succumbed to rabies, had been showing severely deranged behaviour for 3–4 days at the same tourist lodge in Madikwe. It had chased staff and guests around the lodge, torn sofas to shreds, chewed on furniture and roamed inside the lodge buildings, fortunately without direct human contact, until it was shot at the nearby waterhole on 26 January 2015. Subsequent to this, a carcass of a spotted hyaena found dead at a waterhole near the tourist lodge on 27 February 2015 was submitted for testing.On 26 December 2014, a wild dog was found biting at light bulbs in a tourist lodge in Madikwe Game Reserve . It was Half of the brain from the carcasses of the first male wild dog , the second wild dog (ARC-OVI reference number# 66/15) and the spotted hyaena (ARC-OVI reference number# 113/15) were submitted on ice to the OIE Rabies Reference Laboratory at the ARC-OVI to exclude lyssavirus infection. Immunohistochemistry (IHC) was carried out at the Section of Pathology, Department of Paraclinical Sciences at the Faculty of Veterinary Science, University of Pretoria, Onderstepoort on formalinised, wax-embedded brain tissues from the first wild dog and the hyaena. On receipt of the specimens at ARC-OVI, composite brain smears were prepared and subjected to the direct fluorescent antibody test (dFAT) as described previously  (5’-CAAAGGAGAGTTGAGATTGTAGTC-3’)647–666 primers  , sequenced bidirectionally on an ABI3700  and consensus nucleotide sequences of both the forward and reverse sequences were generated using algorithms in MEGA 4  gene. In brief, Trizol-extracted viral ribonucleic acid (RNA) from original RABV-infected brain tissues (approximately 200 ng of tissue) and freeze-dried infected mouse brain material  and wild carnivore species, such as black-backed jackal (C. mesomelas), bat-eared fox  and side-striped jackal (C. adustus).Lyssavirus antigen was confirmed in the brain tissues of the two adult wild dogs and the hyaena using dFAT and for the first wild dog and hyaena using IHC. Typical apple-green fluorescence virus–infected particles were observed in 100% of the field of the composite brain smear and in addition, antigen was observed immunohistochemically in glandular epithelium of the mandibular salivary gland of the first wild dog (889/14) see . The secNucleotide sequencing and phylogenetic analysis of the wild dog RABV partial nucleoprotein gene sequences together with other previously characterised domestic dog and wildlife RABV sequences from South Africa confirmed that canid rabies variant was circulating in these wild dogs . A 100% Rabies trends in the North West province of South Africa over a 10-year period 2005–2015) showed that significant numbers of cases, for example, 44.4% (2007), 58.8% (2012) and 18.3% (2014) of the total annual confirmed animal cases, were positive wildlife cases as recorded at the OIE Rabies Reference Laboratory at the Onderstepoort Veterinary Institute. Whereas the RABVs from wildlife species such as jackals and hyaena in 2007 were of the mongoose rabies biotype, those in 2012 and 2014 were of the canid rabies biotype (Onderstepoort Veterinary Institute records), highlighting the biological variations within the classical RABV variants found in southern Africa and the occurrence of spillover events between wildlife and domestic host species  and wildlife  viruses . This stIn Madikwe Game Reserve in 1997  occasionally dig holes under the fence at Madikwe, although these holes are usually closed by park staff within 24 h of initiation.In South Africa, ‘predator proof’ fencing around small reserves has been very effective at keeping wild dogs confined inside the reserves, provided the fence is properly maintained, but such fencing has also been reported not to be 100% effective , during six years of research (project field data of RH-W). One of these radio-collared individuals attempted unsuccessfully to regain access into Madikwe despite prolonged movements along the boundary fence. There is therefore a low probability that jackals and/or domestic dogs gain access into Madikwe through the perimeter fence.Only 2 out of 39 radio-collared jackals managed to pass through the fence to the outside of Madikwe, using holes made by aardvarks . In this respect, where wild carnivores like lions, hyaenas or wild dogs have made substantial-sized kills, jackals are the most frequently observed mammalian scavengers . In Etosha National Park, jackals are the most frequently observed scavengers at anthrax-confirmed carcasses . A small number of bat-eared foxes were also recorded in the north-eastern region of the park. Aggressive interactions between wild dogs inside the park and canid species outside the park may occur through the fence but had not been recorded up to 2016.Wild dogs usually engorge themselves on fresh carcass kills, mostly kudu in Madikwe; they then lie bloated a little distance from the carcass remains. Although jackals are generally wary of wild dogs with wild dogs often chasing jackals from kills (field observations of RH-W), the disorientation and aggressive behaviour of jackals in the terminal phases of rabies may encourage close physical contact interactions between such rabid jackals and wild dogs.A factor of likely significance in the period prior to the 2014 wild dog rabies outbreak in Madikwe was the effect on the wild dog behaviour of the chemical contraception applied by the park in 2014 as a result of their rapidly expanding population. Wild dogs numbered 34–36 in February 2014 and 26 in September 2014, at which time contraception was introduced. Their numbers thereafter dropped to 13 when counted on 16 February 2015 because of the subsequent rabies outbreak (field data of RH-W). The reproductive intervention had resulted in fragmentation of the northern and southern packs and creation of smaller groups, which appeared to increase antagonistic encounters between these splinter groups. These aggressive contact encounters would likely have exacerbated intra-specific, wild dog rabies transmission. Mills , 1993 ouBetween 2008 and 2010, the Madikwe jackal population was drastically reduced as a consequence of the high lion numbers and densities. The latter resulted in significant opportunistic lion predations on jackals and also an inability of jackals to access lion kills, which in turn were devoured within a short space of time. Lion numbers were reduced steadily between 2010 and 2015 as a result of management initiatives. Jackal numbers are positively favoured when lion numbers are low and when the percentage of lions under 2 years in the population is small, as young lions have been regularly recorded to kill jackals .On 15 of January 2015, a video was recorded of a wild dog male eating a dead wild dog (data of RH-W), highly likely at that time to have died of rabies. The last wild dog of the larger pack, which was the second positively diagnosed wild dog case, was shot and submitted to Onderstepoort on 26 January 2015.The devastating effect of rabies on the wild dog population in Madikwe during the latter half of 2014 was not paralleled in the jackal population (data of RH-W), although data suggest that jackal breeding pairs declined by 15.7% between 2013 and 2014 (pre-rabies outbreak) and by a further 6.2% between 2014 and 2015. In this respect, severe mange, small jackal litter sizes, poor jackal pup survivals and a continued lack of good jackal representations at large carcasses and lion kills between 2013 and 2015 implicate the progressively worsening drought conditions and associated food deprivations as pivotal factors in the poor performance of jackals during this period. Although it was not possible in 2014 to confirm whether all adult jackal pairs associated with pup mortalities were alive after the loss of some or all their pups, some jackal pairs were recorded in their territorial areas after these after the death of these pups. These data challenge rabies as an implicating factor in jackal population mortalities during the latter half of 2014, when rabies probably peaked in the Madikwe wild dog population. This hypothesis is further supported by the observed increase in the numbers of jackals reported at large carcasses during 2016, which suggests that jackal pup survival rates from 2015 jackal litters had improved. The increased availability of food from the extraordinarily large amount of ungulate drought-related mortalities that were present in the field during the latter half of 2015 would have favoured jackal pup survival. The quandary with respect to the origins of the rabies outbreak and as to whether there is a possibility that rabies is present and maintained in certain wild species populations as reservoir hosts within Madikwe needs further investigation (field data and interpretation of RH-W).Spread of rabies amongst members of a spotted hyaena clan as well as inter-clan spread depends on the number of clans, clan size, their density in an area and on the ranks of the affected animals, which would affect behaviour and potential contacts with other hyaenas. In general, widely distributed clans or clans with few members would show slow intra-clan and inter-clan spread. Spotted hyaenas rarely meet with all clan members and individuals tend to be solitary or in small groups. They are more territorial than wild dogs: they scent-mark and defend their territories from other clans Mills . An unusIn February 2015, the remaining five adult wild dogs and eight pups in Madikwe were vaccinated against rabies but six pups still died, although none were tested for rabies. Wild dog pups in Kruger National Park have been recorded to have a high mortality rate, many because of unknown cause but several by lion predation Mills . The 201As human populations increase around reserve borders, the risk to wild dogs which venture outside or escape from parks is also likely to increase. Humans in game parks are at considerable risk of contact with rabid carnivores because several, as in this situation, are shot within tourist lodges or camps or in close proximity to them  genes from virus isolates of four wild dogs  indicated that a viral variant common amongst domestic dogs was responsible for RABV infection in wildlife carnivores in Kenya and Tanzania  of RABV within 1.0–2.5 min (Kaplan, Wiktor & Koprowski Personnel and laboratory employees should treat all fresh samples from suspect rabies cases as potentially rabies-infected and when appropriate throughout the dFAT procedure, as RABV is not inactivated with acetone fixation (Jarvis, Franke & Davis In conclusion, in order for effective control of rabies outbreaks in domestic mammals, non-endangered and endangered wildlife and humans, the epidemiology of RABV infection in these species and in varying circumstances requires ongoing intensive study. This implies that study of predator interactions, movements, behaviour, food sources, competition and the dynamics which alter with human interventions and natural environmental changes within the confines of game parks should be ongoing. Close monitoring of species numbers and submission of material for diagnosis when carcasses are found or when animals are euthanised as a result of aberrant behaviour are essential. Because of its multi-host nature, RABV is an important threat to ecologically threatened animal populations such as the wild dog. RABV is a pathogen that can be effectively controlled or eliminated at the primary host animal source by vaccination. Parenteral vaccination is key to the potential elimination of some viral diseases from domestic dogs and cats and further mitigates transmission of rabies and other pathogens to both wildlife and humans. The diligent ongoing implementation of suitable, effective vaccination of wild carnivores against RABV for continued prevention of the disease and the preservation of these species, although controversial, is recommended.


Mildew damage is a major reason for chestnut poor quality and yield loss. In this study, a near-infrared hyperspectral imaging system in the 874–1734 nm spectral range was applied to detect the mildew damage to chestnuts caused by blue mold. Principal component analysis (PCA) scored images were firstly employed to qualitatively and intuitively distinguish moldy chestnuts from healthy chestnuts. Spectral data were extracted from the hyperspectral images. A successive projections algorithm (SPA) was used to select 12 optimal wavelengths. Artificial neural networks, including back propagation neural network (BPNN), evolutionary neural network (ENN), extreme learning machine (ELM), general regression neural network (GRNN) and radial basis neural network (RBNN) were used to build models using the full spectra and optimal wavelengths to distinguish moldy chestnuts. BPNN and ENN models using full spectra and optimal wavelengths obtained satisfactory performances, with classification accuracies all surpassing 99%. The results indicate the potential for the rapid and non-destructive detection of moldy chestnuts by hyperspectral imaging, which would help to develop online detection system for healthy and blue mold infected chestnuts. Chestnuts, a Chinese specialty, are nutritious and contain plenty of starch, protein and fat. Chestnuts are widely planted in China and their production is increasing year by year . HoweverChestnut mildew is a major cause of chestnut loss ,6. The rHyperspectral imaging combines imaging and spectroscopy technology. Within hyperspectral images, spectral information and image information can be simultaneously extracted from target objects ,9,10. HyArtificial neural networks (ANNs) are computing systems inspired by biological neural networks. ANNs accomplish tasks by analyzing examples and they are widely used where problems are difficult to be expressed in a traditional computation algorithm using rule-based programming. The basic units of ANNs are artificial neurons, analogous to the axons of the biological brain. Signals can be transmitted through the connection between neurons. The receiving neurons can process the signal and then transmit the signal to its downstream neurons. Generally, neurons are organized in layers, where different kinds of transformations of inputs are performed. Nowadays, ANNs have been applied to various fields, such as computer vision, machine translation, speech recognition, social network filtering and many other domains ,23,24.Analyzing the hyperspectral data with artificial neural networks is an effective way to build a classification model. Sun et al. built a set of lettuce leaf nitrogen level identification models based on the hyperspectral image technology and the extreme learning machine (ELM) pattern recognition method. The training time and classification accuracy rate of ELM model were 0.062304 s and 100%, respectively . Zhou etThis work aimed to explore the feasibility of using spectral information obtained from hyperspectral images combined with neural networks, including ELM, BPNN, evolutionary neural networks (ENN), general regression neural network (GRNN) and radial basis neural network (RBNN) models to detect oil chestnuts infected by blue mold.The oil chestnut samples were collected from Anji . Firstly, the chestnuts were cleaned and air-dried. Next, the samples were divided into two groups, healthy chestnuts and healthy chestnuts to be infected. To obtain enough infected chestnuts, some chestnuts infected by blue mold were placed among normal chestnuts under 85% humidity and 25 °C for one week. There were 264 healthy chestnuts and 264 infected chestnuts prepared for this study. Then, these chestnuts were randomly split into the calibration set and prediction set at the ratio of 2:1, with 176 healthy samples and 176 infected samples in the calibration set and 88 healthy samples and 88 infected samples in the prediction set. The entire sample set  was imaged in multiple batches  because of the size limitations of mobile platform.The hyperspectral imaging system was provided by the Zhejiang University College of Biosystems Engineering and Food Science. The system was installed at a black sealed box. It could acquire 256 spectral image bands in the region of 874–1734 nm with spectral resolution of 5 nm. As shown in A black plate with low reflectance was used as background in the study. With this black plate, chestnuts could easily be isolated from the background. During the image acquisition, the height between the camera and the platform was 26.5 cm, the moving speed of the platform was 23.5 mm/s, and the exposure time of the spectral camera was 2 ms.rawI) was calibrated by two reference standards. Black reference image (darkI) was obtained by covering the lens with lens cap whose reflectivity was about 0%. A pure white Teflon board which had a high reflectivity (about 100%) aligned with the lens to obtain a white reference image (whiteI). The calibrated image (cI) could be calculated as follow [In this study, the accuracy of reflectance would be influenced by the illumination source and the sensitivity of detector. The original image (s follow :(1)Ic=IrIn this study hyperspectral image data were acquired in the 874–1734 nm spectral region. After the removal of background, the entire sample region of each chestnut was defined as a ROI, and pixel-wise spectra within the ROI were extracted. Due to the obvious random noises at the beginning and the end of spectra, only the spectral range of 975–1646 nm was studied. The pixel-wise spectra should be preprocessed to minimize the random noise before analyses. Taking the edge effect into account, we substituted a constant (0) for the edge of ROIs. Wavelet transformation using the wavelet function Daubechies 7 with decomposition level 3 was used to reduce the random noises ,29. AfteThe Successive Projection Algorithm (SPA) is a forward-loop variable selection method that can extract valid information from the redundant spectral information, minimize the collinearity between spectral variables and improve the modeling conditions of multiple linear regression (MLR) model. SPA was applied to reduce the dimensionality of data in spectral domain of chestnuts during the study .The back propagation neural network (BPNN) is a multi-layer feed-forward artificial neural network with back-propagation algorithm. BPNN has the capability of large-scale parallel distributed processing with distributed information storage method. BPNN also has a strong self-adaptability to the environment and the learning ability of the outside world, as well as strong robustness and fault tolerance. BPNN is usually composed of an input layer, hidden layer and output layer. The layers are connected to each other, but nodes in each layer are not connected between each other. The number of its input layer neurons usually takes the value of the dimension of the input vector. And the number of output neurons usually takes the value of the dimension of the output vector. There is no definite standard for choosing the number of hidden layer neurons, so it is often decided by traversing within the set range ,31,32.The topology structure of BPNN model is shown in The evolutionary neural network (ENN) is a new neural network model based on two intelligent branches, the evolutionary computation and the neural network. One of the main features of the ENN model is its adaptability to dynamic environments. This adaptive process is realized by three levels of evolution, including the evolution of connection weights, network structures and learning rules . Figure The extreme learning machine (ELM) is a kind of single-hidden layer feed-forward neural network (SLFNN). Only the output weights are required to get the global optimal solution. ELM has fast learning ability, strong generalization ability and produces the only optimal solution. ELM algorithm can be used for discriminant analysis or regression analysis. In the practical application the number of neurons in the hidden layer needs to be determined, and it is currently determined by updating the parameter value to get optimum solution ,34,35.The topology structure of the ELM model is also shown in The general regression neural network (GRNN) is a kind of radial basis function (RBF) network developed by Specht in 1991. It is a powerful regression tool with a dynamic network structure, whose training speed is extraordinarily fast. Its simplicity of the network structure and the implementation endows GRNN popularity in a variety of fields like image processing ,37.The radial basis neural network (RBNN) originates from the usage of radial basis functions in the solution of real multivariate interpolation problems. As a multilayer perceptron (MLP) they can approximate any regular function. Because of its local behavior and the linear nature of its output layer, it can be trained faster than other MLPs, which makes it useful in various applications. However, the generalization capability of RBNN is normally poor because it focuses too much on the training data set. Hidden-layer neurons represent regions of the input space, which makes the search of the appropriate number of hidden-layer neurons become a difficult task ,39. SpreTo explore qualitatively discrimination of healthy and moldy chestnuts, principal component analysis (PCA) was applied. PCA was conducted in two ways in this study, including PCA scores image and PCA scores scatter plots. PCA scores image were obtained by applying PCA on the hyperspectral images. The background was firstly removed from the hyperspectral images using the method described in The spectral data in the 975–1646 nm range with 200 wavelength variables was studied. The average spectra of healthy chestnuts and moldy chestnuts with standard deviation (SD) of wavelengths at peaks and valleys are shown in PCA scores image could be depicted according to pixel-wise PCA scores. As seen from It can be seen from PCA was performed on the average spectra data for qualitative discriminative analysis of healthy chestnuts and moldy chestnuts. The first three PCs explained over 99% of the total variance. To represent the distribution of chestnuts in The spectral data obtained by hyperspectral imaging systems usually have a large data volume and contain a lot of irrelevant information like redundant, collinear and background information. The existence of irrelevant information will not only affect the extraction of useful information, but also increase the data processing burden, which is likely to cause instability in the model and thus a poor performance. Meanwhile, the large amount of computation places a high requirement on computer hardware. The characteristic wavelengths are the wavelengths containing the useful information selected from a large number of wavelengths by removing the redundant and collinear data and wavelengths representing the background signal. The selection of characteristic wavelengths can reduce the inputs, simplify the model and improve the model performance.In this study, SPA was adopted to select the optimal wavelengths. It can be seen from In order to evaluate whether the infected chestnuts could be detected by hyperspectral imaging, BPNN, ENN, ELM, GRNN and RBNN models were built for comparison. There were 264 healthy chestnuts and 264 blue mold infected chestnuts. Each variety was split into a calibration set and a prediction set at a ratio of 2:1.In order to obtain a better classification results, the parameters of BPNN, ENN, ELM, GRNN and RBNN, which were the number of neurons  or spread value (GRNN and RBNN) of the hidden layer, should be optimized first. The parameters were selected according to the classification accuracies of the calibration set and prediction set of each model. The changing trend of classification accuracy of calibration set with different parameters of classification models based on optimal wavelengths is shown in For models using full spectra, the accuracy of the calibration set and the prediction set of BPN and ENN models all reached 100%. The accuracy of the calibration set of ELM and RBNN models was the same as for the BPNN and ENN models, reaching 100%. However, the accuracy of the prediction set of ELM and RBNN models was slightly lower than that of the BPNN and ENN models, which only surpassed 80%. The accuracy of the calibration set and the prediction set of GRNN model based on full spectra was much lower than that of other models, with 71.31% accuracy for the calibration set and 55.68% accuracy for the prediction set. For the models using optimal wavelengths, the accuracy of the calibration set of BPNN, ENN, ELM and RBNN models all reached 100%. However, the prediction accuracy of ELM model only reached 81.25%, while the prediction accuracy of three other models  all surpassed 96%. The BPNN, ENN and RBNN models performed the best among the five models. The prediction result of the ELM model based on optimal wavelengths was inferior to that of the BPNN, ENN and RBNN models with a prediction accuracy of 81.25%. The GRNN model based on optimal wavelengths showed the worst results among the five models, with 62.5% accuracy for the calibration set and 56.82% accuracy for the prediction set. The selection of optimal wavelengths reduced the volume of data tremendously, and improved the speed of modeling and prediction, especially for the BPNN, ENN and RBNN models. Comparing the classification results of the models based on full spectra with the models using optimal wavelengths in A new method for discrimination of moldy chestnuts was proposed in this study. A hyperspectral imaging system with a wavelength range of 874–1734 nm was applied to identify moldy chestnuts. PCA score visualization images were depicted to distinctly show the differences between healthy and moldy chestnuts. In order to reduce the training time of models, 12 optimal wavelengths were selected by SPA. Artificial neural networks such as BPNN, ENN, ELM, GRNN and RBNN were used to build models based on full spectra and optimal wavelengths. Parameter optimization was conducted in the five models in order to improve the classification accuracy. All models except for the GRNN model achieved satisfactory results. Among all the models using full spectra and optimal wavelengths, the BPNN and ENN models obtained the highest accuracy with all classification accuracies reaching above 99%. The results showed the feasibility of using hyperspectral imaging combined with an optimal wavelengths selection method and artificial neural networks to discriminate moldy chestnuts.Further studies are expected to conduct more research on how to distinguish moldy chestnuts with different varieties of mold species using hyperspectral imaging system. More chestnut samples from different regions should be taken into account in order to improve the robustness of models. Besides, more study should be conducted on how to improve the universality of the discrimination models.


Memories can be initiated by coincident events, as occurs in classical conditioning, where the NMDAR can act as a molecular coincidence detector. Binding of glutamate and glycine, together with depolarization of the postsynaptic cell membrane to remove the Mg2+ channel pore block, results in NMDAR opening for Ca2+ conductance. Accumulating evidence has implicated both force-from-lipids and protein tethering mechanisms for mechanosensory transduction in NMDAR, which has been demonstrated by both, membrane stretch and application of amphipathic molecules such as arachidonic acid (AA). The contribution of mechanosensitivity to memory formation and consolidation may be to increase activity of the NMDAR leading to facilitated memory formation. In this review we look back at the progress made toward understanding the physiological and pathological role of NMDA receptor channels in mechanobiology of the nervous system and consider these findings in like of their potential functional implications for memory formation. We examine recent studies identifying mechanisms of both NMDAR and other mechanosensitive channels and discuss functional implications including gain control of NMDA opening probability. Mechanobiology is a rapidly growing area of biology with many important implications for understanding form, function and pathology in the nervous system.An increase in post-synaptic Ca In addition, the NMDAR plays an important role in intra-neural network activity, where propagating and feedback circuits also can undergo memory related plasticity  enabled investigations into the essential role of NMDAR in memory formation. In comparison to control animals, APV-treated animals failed to form spatial memories of platform location. Control studies confirmed that APV did not affect visual or other sensory systems and the effects were specific to the formation of memory . In addiThe NMDAR ion channel and the subunits contributing to the functional ion channel (see below for subunit details) have been localized to postsynaptic afferent sites as well as to internal excitatory circuits of the hippocampus and amygdala, by electrophysiological and electron microscopy studies. Recurrent circuits necessary for spatial memory pattern recognition in the hippocampus CA3 regions are mediated by NMDA receptors . Likewisin vitro and in vivo models of synaptic plasticity. A leading model for this process is LTP of excitatory currents at potentiated synapses. LTP can be induced by high frequency (100 Hz) stimulation of afferent synapses  and traumatic brain injury (TBI) . Patholo regions . These pvia conditional Ca2+ signaling  receptors, provide a satisfying explanation for the function of this ligand-gated ion channel as a molecular coincidence detector in LTP and memory . This is because upon removal of the large inositol phosphate head by PLC a small molecule of DAG is left in the inner leaflet of the bilayer that causes the bilayer to curve locally. The change in the local membrane curvature has been proposed to induce membrane stress large enough to open TRPC6 ion channels, for example  molleagues ; (iii) Nlleagues . Additiolleagues . More relleagues .2+ entry and axon formation (The micro-anatomical structure of neurons can undergo regular physical transformation. Micro- structural plasticity is proposed as a major physical representation of memory . During ormation . These pIf NMDAR mechanosensitivity leads to modulation of behavior, a key candidate mechanism is modulation of memory systems in the brain. Moreover, pathology in these mechanisms may lead to altered memory functions as is observed in memory related disorders including PTSD, dementias and TBI. NMDAR mechanosensitivity may be related to memory function through several mechanisms. Early studies by Atwell and colleagues  suggesteLJ and BM planned and wrote the manuscript. AB prepared the figure. All authors edited the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Adverse food reactions (AFRs) are defined as abnormal responses to an ingested food or food additive. Diagnosis and treatment of AFRs consist of the complete elimination of these ingredients in the dietary trial. Previous studies have demonstrated the presence of undeclared ingredients in commercial limited-antigen dry food diets that can compromise the results and efficacy of dietary elimination trails. The aim of this study was to assess a selection of commercial canine and feline dietetic limited-antigen wet foods for the potential cross-contamination of animal proteins from origins not mentioned on the label.Eleven canine and feline dietetic limited-antigen wet foods  were analyzed by polymerase chain reaction (PCR) to detect the presence DNA of animal and vegetal origins. PCR analysis confirmed the contamination of 6 of the 11 (54.5%) limited-antigen wet diets with undeclared animal protein. One of these 6 diets was solely composed of animal protein sources completely unrelated to those declared on the label. None of the foods containing horse meat or fish were contaminated, and neither were the vegetarian or the hydrolyzed food products. Moreover, the results show that had zoological class primers only been used to check for cross-class contaminations, as are generally used in the pet food industry for in-house checks, the apparent contamination rate would have been significantly underestimated: less than 20% (3/11), instead of the actual rate of 54.7% using species-specific primers.This study reveals a high rate of cross-contamination in dietetic limited-antigen wet canine and feline foods, as previously described for dietetic dry limited-antigen foods (reported to be more than 80%). These results add new fuel to the discussion about the potential causes underlying the failure of elimination diets, since animal protein contaminants may actually be present in the commercial dietetic limited-antigen diets. AFRs may therefore occur as a result of inadequate practices in the pet food industry. Adverse food reactions (AFRs) are defined as abnormal responses to an ingested food or food additive , 2. WhenEleven canine and feline antigen-limited wet diets (10 novel protein diets and 1 hydrolyzed diet) produced by 5 different pet food manufacturers  were obtained from veterinary clinics. All of the products used in this study were declared as dietetic complete pet food, specifically intended for particular animal nutritional purpose of reduction of ingredients and nutrient intolerances, according to the European law 2008/38/EC . For eacEach pet food product was randomly assigned a unique three-digit sample identification number. Samples were then submitted to the polymerase chain reaction (PCR) to identify DNA belonging to 3 zoological classes . Based on these results, PCR analyses for specific animal species  were performed according to the zoological classes testing positive. The limited-antigen diet composed of vegetal protein only was also tested using PCR to identify vegetal protein origin.To extract DNA from pet food samples, the DNeasy Blood and Tissue Kit  was used following the manufacturer’s instructions. Negative controls (water) were included in each extraction. DNA concentration was determined by spectrophotometry .. For horse, a specific real time PCR assay was run in a 7300 Real-Time PCR System . The PCR amplifications were performed in an ABI 2720 thermocycler . Minisequencing was performed on an ABI 310 Genetic Analyzer . All amplification protocols were performed following the published protocols , and subsequently for DNA from specific animal species as well as for DNA of vegetal origin. Table The elimination diet is currently the most important and reliable diagnostic test for evaluating and diagnosing AFRs in dogs and cats . The tesIndeed, the high prevalence of AFRs in dogs and cats is an important reason why allergen management should become an integral part of the food safety management system in the pet food industry. In canine populations, AFRs were diagnosed in 1.69% of dogs presented to a veterinary teaching hospital in a single year . In catsThe possibility of intentional cross-contamination of either feed materials or final products is unlikely considering the similar or sometimes higher cost of the undeclared animal species identified in our samples, such as the horse protein found in sample n.7 which declared duck as its unique animal protein source. Intentional cross-contamination might occur in situations of limited availability of some selected protein sources, such as duck, for which availability cannot always be guaranteed for pet food production with high meat inclusion rates.In order to ensure a high level of feed safety and to improve transparency, provisions should be added as an integral aspect of good practice. Although every effort could be taken by the pet food industry to eliminate the risk posed by the unintended presence of food allergens, in many businesses it is virtually impossible to produce a zero risk product. In many manufacturing premises, production lines dedicated to the manufacture of a single product are not always feasible or practical for economic or logistical reasons. Therefore, the cleaning of shared equipment, processing lines and the local environment becomes a key element in allergen control. Manufacturers are presently required to define the cleaning procedures and cleaning schedules appropriate for their facilities. But they are not required to ensure or to demonstrate that the cleaning programme is effective and performing properly through the use of specific and accurate analyses, such as PCR-food analysis. Multiple lines are more frequently used for the production of commercial dry pet food than for the production of wet food. This could justify the lower contamination rate of our wet samples (54.5%) compared with that for dry foods analysed by other authors, described as being over 80% . Our metOne of the most interesting results obtained in our study using the species-specific PCR was that diets with protein sources derived from horse and fish (tuna) were always clean . These data might imply that the different feed material production techniques affect the potential for cross-contamination by other animal protein sources; moreover, they highlight and shift the focus from the pet food manufacturers to the suppliers of feed materials . It seems that when separate lines are required for the slaughter, transformation and distribution of animal products (as is the case for horse meat and fish), the final by-products are also cleaner. In the past, some authors also focused their attention on lipid sources of animal origin. In the present study, the animal lipid sources declared on the label were never found to be species-specific or zoological class PCR positive . It woulThe results of this study are very important for pet food producers of limited-antigen diets. They indicate the need for improvements to be made in the selection of feed material suppliers and the type of checks installed throughout the production chain before final products are introduced onto the market. Importantly, our results indicate that the type of checks used should involve PCR analyses for species-specific classes and not just zoological class.In conclusion, the high incidence of contaminations found in the antigen-limited wet diets analyzed in our study confirm the severity of the problem as previously highlighted for dietetic limited-antigen dry diets . These r


The crosstalk between cancer cells and host cells is a crucial prerequisite for tumor growth and progression. The cells from both the innate and adaptive immune systems enter into a perverse relationship with tumor cells to create a tumor-promoting and immunosuppressive tumor microenvironment (TME). Epithelial ovarian cancer (EOC), the most lethal of all gynecological malignancies, is characterized by a unique TME that paves the way to the formation of metastasis and mediates therapy resistance through the deregulation of immune surveillance. A characteristic feature of the ovarian cancer TME is the ascites/peritoneal fluid, a malignancy-associated effusion occurring at more advanced stages, which enables the peritoneal dissemination of tumor cells and the formation of metastasis. The standard therapy for EOC involves a combination of debulking surgery and platinum-based chemotherapy. However, most patients experience disease recurrence. New therapeutic strategies are needed to improve the prognosis of patients with advanced EOC. Harnessing the body’s natural immune defenses against cancer in the form of immunotherapy is emerging as an innovative treatment strategy. NK cells have attracted attention as a promising cancer immunotherapeutic target due to their ability to kill malignant cells and avoid healthy cells. Here, we will discuss the recent advances in the clinical application of NK cell immunotherapy in EOC. Most EOCs are diagnosed at an advanced stage, which accounts for the high mortality rate associated with this disease. The new World Health Organization (WHO) Classification of Ovarian Cancer takes the recent findings on the origin, pathogenesis and prognosis of different ovarian cancer subtypes into account. The tubal origin of hereditary and some non-hereditary high-grade serous cancers is mentioned in contrast to the previous theory of mesothelial origin of tumors while seromucinous tumors represent a new entity  with mAbs  or with Given the results of GOG-0218  and ICONAnother promising approach is the infusion of antitumor lymphocytes that were previously engineered with chimeric antigen receptors (CARs). These studies are mainly focused on cytotoxic T cells but recently, these technologies have also been applied to NK cells. In a recent study, the effect of the chemotherapeutic agent cisplatin in association with a CAR-based immunotherapeutic approach was evaluated to improve the clinical efficacy of ovarian cancer therapies. In particular, a lentiviral vector encoding a third-generation anti-CD133-CAR was generated and transduced in NK92 cells. This combined clinical approach led to a strong killing effect against ovarian cancer stem cells . EffortsIn high-grade serous ovarian carcinomas, the most common histological type of ovarian cancer, almost 15% of women harbor germline BRCA mutations . A recenCurrent efforts for a NK cell based therapy are mainly based on strategies that manipulate the function of inhibitory receptors. In this context, it has been recently demonstrated that ascites-associated NK cells can also express high levels of PD-1, with this expression potentially impairing the antitumor activity of these innate effectors . These fFuture research is needed to clarify the effects that checkpoint inhibitors have on the NK cell response and the potential to enhance adoptive NK cell immunotherapy in ovarian cancer .A combination of these different techniques of NK cell-based immunotherapy hold great potential and may represent an effective weapon against ovarian cancer after primary cytoreductive surgery and adjuvant chemotherapy.Based on the evidence of the high frequency of persistent tumors at the completion of primary postoperative chemotherapy and the high relapse rate for women with advanced EOCs treated with standard surgery/chemotherapy, there is a clear medical need for identifying a management strategy that results in improved outcomes. Further studies are needed to provide new insights on the immunological mechanisms involved in the development of certain forms of ovarian cancer, which will be crucial for the development of new immunotherapeutic strategies .The recent evidence for a strong prognostic effect of ER in a large proportion of newly diagnosed EOC offers a therapeutic target for a very cost-effective precise medical approach.Increasing our understanding of the mechanisms of NK cell activation, including those regulated by different molecular checkpoints and hormone receptors, and the identification of new tumor biomarkers will provide a firm basis for how to optimize NK cell reactivity against cancer. This knowledge can be applied to the development of an optimal design for cancer immunotherapy by targeting NK cells either alone or in combination with other therapies.


We estimated the availability of the injectable antimicrobial drugs recommended for point-of-care treatment of gonorrhea and syphilis among US physicians who evaluated patients with sexually transmitted infections in 2016. Most physicians did not have these drugs available on-site. Further research is needed to determine the reasons for the unavailability of these drugs. Neisseria gonorrhoeae  are on the rise in the United States. Compared with the incidence in 2013, the numbers of reported gonorrhea (+75%) and syphilis  cases in 2017 were dramatically increased .The recommended first-line treatment for uncomplicated gonorrhea is intramuscular ceftriaxone (250 mg) and for primary and secondary syphilis is intramuscular penicillin G benzathine (2.5 million units)  of office-based physicians indicated that they evaluate patients for STIs in their offices. Of these, 77.9%  reported not having penicillin G benzathine available on-site and 56.1%  reported not having ceftriaxone. Access to both of these drugs was generally higher in the South . PhysiciOur nationally representative analysis demonstrates that most office-based physicians who provide STI services reported not having on-site access to the recommended injectable medications for gonorrhea and syphilis management. The PCMH designation has been noted as an indicator for quality of care by the American Medical Association (We provide a national estimate of the percentage of US physicians treating patients with STIs that report not having the recommended antimicrobial drugs on-site for same-day treatment of gonorrhea and primary and secondary syphilis. In light of our findings, we believe that future research should focus on determining the factors preventing physicians from providing these medications on-site. The costs of obtaining and carrying these medications, as well as issues of storage and shelf life, should be explored to determine if these factors are barriers. In addition, the implications of prescribing alternative treatments or delaying care in situations when medications are not readily available on-site should be further explored. Mitigating the lack of medication availability to treat these infections will help public health officials stop the rise in STI disease.


Giardiasis has a global distribution and it is a common cause of diarrhea in both children and adults and is transmitted via the fecal-oral route through direct or indirect ingestion of cysts. The laboratory diagnosis of Giardia spp. is mainly based on demonstration of microscopic cyst or trophozoite in stool samples but several immunological-based assays and molecular methods are also available for giardiasis diagnosis. The aim of this study was to conduct a review of the applied methods in medical laboratory and to highlight pitfalls and challenges of them for diagnosis of giardiasis. In this article we have evaluated the Giardia diagnostic methods with a broad review of literature, electronic databases and books. The search has covered the articles and some textbooks that have published up to 2018.It has been concluded that traditional microscopy combination with stool concentration method should still be held in the routine medical laboratory due to economical and high sensitivity and immunological-based assay and molecular methods which are recommended to use as a complementary test to the traditional technique. Giardiaduodenalis  is one of the most prevalent intestinal protozoan flagellate of the human. The life cycle of Giardia species is simple and it is included of two active trophozoite and cystic forms. The etiological agent of Giardiasis, This parasite transmits via fecal-oral route through direct or indirect ingestion of infectious cysts. The incubation period varies from 9 to 15 days after ingestion of cysts. Symptoms of infection are varied from the absence of symptoms to acute watery diarrhea, nausea, epigastric pain and weight loss . Giardia infection is higher in developing countries. More than 200 million cases of giardiasis are annually diagnosed worldwide. Since 2004, Giardia has been included in the "neglected diseases initiative" by World Health Organization  and culture for diagnosis of Giardiasis.Giardia", "diagnosis", "immunodiagnosis", "molecular" have been used as a panel of keywords. The search restricted to English and Persian languages and the references of selected papers were checked for more accuracy. Electronic searching was performed in the international databases such as ISI Web of Science, PubMed, Scirus, EMBASE, Scopus, Science Direct and Google Scholar. The national databases searching were Iran Medex, Iran Doc, Magiran and Scientific Information Database. The following keywords: "Diagnostic methodsFecal microscopy examinationGiardia spp. in fecal samples is considered as the gold standard method for the diagnosis of giardiasis. This method is performed to detecting cysts and trophozoites. The sensitivity of microscopy techniques depends on using direct or concentration methods, the number of examined fecal samples and employment of professionally trained persons .The diagnosis of giardiasis in most cases is mainly confirmed by stool examination. Fecal suspension in physiological salt solution (0.85 NaCl) or fixation in sodium acetate–acetic acid–formalin (SAF) is used to prepare wet mounts in order to the observation of Examination of direct wet saline preparation of a fresh stool specimen allows motile trophozoites to be seen, but in stained and SAF preparation smears the trophozoites will be non-motile. If diarrhea stool sample containing trophozoite left too long without fixations or preservatives solution, the organisms tend to degeneration, thus preventing has been recommended for sample transfer and protection of the typical trophozoite morphology. A number of commercial kits with preservative solutions are available or can be made manually. The most commonly used preservation kit contains of 10% buffered formalin, polyvinyl alcohol (PVA), merthiolate-iodine-formalin, and SAF solution , 13. PolIn the asymptomatic individuals and healthy carrier who do not have diarrhea, the cyst stage is more likely to be seen in a fecal sample examination. Fecal suspension in saline or lugol’s solution or in a fixative solution may be used for cyst identification. How many fecal examinations are necessary to detect Giardia cyst by wet mount methods? In these cases, the number of cysts may be low in the fecal specimens, thus the wet mount examination of stool samples may not detect the parasite. It has been recommended for preparation and examination of two or more, even to six wet mounts smear for increasing the chance of finding parasite agents. Also, it must be requested from patient to submit more than one stool samples on consecutive days due to the intermittent shedding of the cyst. Examination of one stool sample will allow the diagnosis of 60 to 80% of infections, two stool samples examination will allow the detection of 80 to 90%, and diagnosis will be over 90% if three stool samples have been examined . HoweverDirect microscopy method has been considered economical and rapid for the diagnosis in the medical diagnostic laboratory.Concentration methodsGiardia cysts may be missed by using wet mounts direct smear. Concentration methods have been designed to separate protozoan cysts and helminthes eggs from excess fecal debris. Flotation and sedimentation are two types of concentration procedure that have been used in the parasitological laboratory. Flotation methods permit the separation of some protozoan cysts and helminthes eggs through the use of a liquid with high specific gravity such as NaCl, NaNO3, ZnSO4 . Zinc sulfate has been recommended as the best saturated solution to detection of Giardia cyst  for this method compared to direct and formalin–ether methods. They suggested that sucrose density gradient is a suitable diagnostic method and it can be used in place of formalin-ether concentration  (Some permanent stains have been used for s method -22. Althe/black) -24.Trichrome is a shorter permanent stain that is simple and well- stained smears in about 45 min to 1 hour. This procedure is of value for staining fresh faecal specimens as well as stool fixed in PVA. In this staining, the background materials stain green or blue-green. The cytoplasm of trophozoites and cyst stain green or greenish-blue, nuclei and nuclear chromatin stain red or red-purple .Culture methodsGiardia spp. in the clinical diagnostic laboratory. Cultivation of Giardia spp. is applied in the research laboratory for many types of studies that require a large number of trophozoite.Although cultivation of human intestinal protozoa is a useful method for detection and diagnostic purpose, routine culture techniques were not established for Giardia spp. is grown in the monoxenic and xenic type of culture system. In monoxenic system, the parasite has been grown in the presence of a single additional flora organism species and in axenic, parasite has been grown in the absence of any other accompanied alive cell (Giardia spp. can be established directly into axenic media. The ive cell . MonoxenGiardia axenic culture is Diamond's medium "TYI-S-33" which modified by Keister DB ( The most common and suitable used medium for ister DB -26.String test (Entero-Test) Giardia trophozoites  may be useful for revealing the hozoites ,27. The hozoites . Some st methods -30.Also, a drop of mucus can be fixed directly on the slide and used for permanent staining .Giardia spp. diagnosis is little known and reported inconsistent. Some researchers have reported that Entero-Test is reliable and superior to stool examination for identification of Giardia spp. in human and dog  as the predominant antibody has been detected in duodenal fluid and saliva samples of infected people. The production of secretary IgA has been developed during active giardiasis, so detection and monitoring this antibody may be a useful tool for serodiagnosis  (A study on p<0.001) . Giardia infection.A variety of assays such as Elisa, IFA, Western blot have been used for the serodiagnosis of giardiasis, but these methods may be problematic as the antibody may be detectable as long times after treatment of acute diseases. Commercially produced kits were not developed for detection of serum antibodies to Antigen detectionSome methods include immunoassay techniques  have been used to detect fecal antigens in both preserved formalin- and fresh stool specimens .G. intestinalis antigens. Using immunoassay kits have been described for simultaneous detection of Giardia and Cryptosporidium or Giardia spp., Cryptosporidium and Entamoeba species antigens in faecal specimens  which presenst in both trophozoites and cysts .target gene sequence which has been used in different molecular studies of Giardia species are genes encoding small subunit (SSU) ribosomal RNA, glutamate dehydrogenase (gdh), triosephosphate isomerase (tpi) and β-giardin genes  or assemblages , the small-subunit of ribosomal RNA (SSU), and triosephosphate isomerase (tpi) genes, showed that emblages , 5. All ectively .Giardia spp., Cryptosporidium,Dientamoeba and Entamoeba histolytica with a high sensitivity and specificity , the sensitivity and specificity for the Nested-PCR in diagnosis of giardiasis was 89.9% and 82.9% respectively (A recent study by Hijjawi ectively .Giardia spp. is one of the most common waterborne parasites that infected human. Cyst stage of this parasite has been identified in surface waters such as rivers, lacks, and ponds. Contaminated food and water to Giardia cysts by the food-handlers can be one of the most important sources of transmission of this parasite to humans (o humans . The maio humans .Giardia infection. Several immunodiagnostic tests of rapid diagnosis of giardiasis have been developed particularly in the last three decades, mainly based on the detection of Giardia antigens in faecal specimens. While to the high sensitivity of these methods (It is well known that no traditional or new methods can detect all cases of  methods , microsc


A 34-year-old man, trader, and married with four wives, otherwise healthy, without any remarkable medical history, was admitted in urology ward in emergency with superficial venous thrombosis of the penis known also as Penile Mondor's Disease (PMD), a rare nosologic entity of the penis associated with pain and an indurated dorsal cord of the root of the penis. The patient receives nonsteroidal anti-inflammatory drugs and coagulation and platelet aggregation inhibitors drugs. Healing and total recovery occur after eight weeks without any complications. Mondor's Disease (MD) is a rare nosologic entity, most often localized in the anterolateral thoracic wall and mammary region. It was first described by Henri Mondor in 1939  and thenA 34-year-old man, married with four women, trader, with no known past medical history, was admitted in emergency in the urology division at Souro Sanou University Teaching Hospital of Bobo-Dioulasso (Burkina Faso) for a painful swelling penis following laborious sexual intercourse. This symptomatology began nine (9) days later with progressive painful swelling at the balanopreputial ring just after sexual intercourse. The patient consulted previously in a private clinic and was given some medication without success, and then he is admitted in emergency in our urology ward because of the generalized involvement of the entire penis with painful oedema. Medical history does not find any recent trauma or use of any medication; no urinary tract symptoms were reported but recent intense sexual intercourse was noted. The patient reported that he is married to four women and regularly having vigorous sexual intercourse activities. Physical examination found an indurated subcutaneous filiform and painful cord, palpated at the dorsal root of the penis with soft swelling of the whole penis .Examination of genito-urinary system was normal. Standard investigations (blood and urine) requested were normal, and blood cell count, urea, and creatinine were without anomalies. Urine analysis does not reveal any infection. Suspicion of penile fracture was made but anamnesis does not find a popping sound or hearing a cracking or losing an erection suddenly during sexual intercourse. Secondary, diagnosis of thrombosis of the superficial dorsal penile vein was made. An ultrasonography coupled with Doppler of the penis was requested and revealed a thrombosis of the superficial dorsal vein of the penis with the presence of the intravascular blood clot thrombosis .To be sure to excluded penile fracture, penile magnetic resonance imaging (MRI) was performed and do not reveal any traumatic lesion of cavernous and spongious bodies .Conservative treatment was done with nonsteroidal anti-inflammatory drugs (NSAID) and acetyl salicylic acid stick (100mg/day) for thrombosis prevention by coagulation and platelet aggregation inhibition. The patient was informed about the necessity of sexual abstinence till disappearance of all symptoms. The evolution was marked by quick and favorable recovery with disappearance of pain in eight weeks, reduction of the swelling, and complete and total recovery of the penis. At six months of follow-up, no evidence of reoccurrence was observed.Penile Mondor's Disease (PMD) by superficial venous thrombosis of the dorsal vein of the penis described the first time by Braun Falco in 1955 is a benign genital condition . It affeMondor's Disease localized in the penis is a rare and benign pathology that the pathogenesis is not yet clear and well known. The diagnosis is based on clinical findings and the investigations by Doppler coupled with ultrasound can contribute to the diagnosis. Conservative therapy with observation or NSAID anticoagulation agent administration and temporary sexual abstinence can occur healing with total recovery. Surgical treatment can be indicated when there is persistence of thrombosis after a long-term medical treatment.


The relatively high incidence of recurrence of bladder cancer is a serious problem in clinical practice. At present, there are no objective microscopic criteria for evaluation of the tendency for local relapse. Besides the phenotypic properties of the tumor parenchymal cells, possible signs in regard to recurrence could also be derived from the peculiarities of the tumor stroma. The stromal reaction, manifested by inflammatory infiltration in the tumor is considered to influence the biological behavior of tumors. Also, a relationship has been reported between the number of eosinophils and the survival of patients. The aim of the present study is to analyze tumor-associated tissue eosinophilia (TATE) and to compare TATE density in the initial foci of age and gender-matched 156 cases of recurrent and non-recurrent bladder cancers; the tumors that have relapsed within six months after removal and contained statistically significant greater numbers of eosinophils in primary cancer sites. These results suggest that TATE may be one of the probable prognostic signs for local relapse of urothelial cancer. Bladder cancer is a common urologic malignancy and has one of the highest recurrence rates . IncreasSome studies support the opinion that eosinophils may have a role in tumor initiation . In coloThe aim of the present study is to evaluate TATE in primary foci of recurrent and non-recurrent urothelial carcinoma of the urinary bladder.A total of 156 cases of urothelial carcinoma were studied on the primary site formalin-fixed parathion embedded tissue sections, stained with hematoxylin and eosin (H&E). Out of them, 78 had a disease recurrence within a period of six months. Their morphological features were compared to a control group of age and sex-matched 78 cases without recurrence for the same period of time. We examined the density of eosinophilic leukocytes per square millimeter of tissue surface.We performed morphometric analysis by using digitally scanned slides via Leica Aperio ScanScope AT2 device  visualized on ImageScope V12.1.0.5029 .Statistical analysis was carried out with SPSS Statistics v.23 using descriptive statistics. Categorical features were summarized with frequencies and percentages. The Mann–Whitney U test and Pearson correlation were used for comparison and estimation of correlations between the density of eosinophilic leukocytes in the stroma of non-recurrent and recurrent bladder cancer. Specificity and sensitivity of density of eosinophilic leukocytes in stroma for distinguishing recurrent disease patients (RP) from non-recurrent disease patients (NP) at three years of follow-up were evaluated with receiver operating curve (ROC) analysis. Diagnostic accuracy of biomarkers was also determined by obtaining the largest possible area under the curve (AUC) in ROC analysis. Kaplan-Meier survival curves and the log-rank test were used to compare the survival differences between groups. Although our study was not powered enough to compare different subgroups, hazard ratios (HRs) and corresponding 95% confidence intervals (CIs) were calculated by Cox regression models. Two-tailed p-values (<0.001) were considered as significant.In the present study, eosinophilic leukocytes were present in the stroma of 66 tumors 42.3%), either as scattered single cells, small groups of cells, or larger clusters. The Mann–Whitney test showed that there are significant differences in the number of eosinophils in the RP and NP group. TATE varied from entirely lacking eosinophilic leukocytes to moderately present and to very high density Figures -3.%, eitherThe density of eosinophilic leukocytes in the tumor stroma varied severely between the RP and NP groups . At the optimal cutoff values of number of eosinophils, the sensitivity was 75.4% and specificity was 78.9%  was calculated as the time from the date of surgery to the date of documented recurrence of the disease. Cancer patients with a high number of eosinophils had no significant differences in DFS when compared with those with a low number of eosinophils Figure .The quantitative assessment of TATE in the tumor stroma shows statistically significant higher TATE in primary urothelial cancer, with a tendency to recur.The present study was designed to evaluate the significance of TATE in the stroma of urothelial cancer. We focused on one of the most peculiar features – the relatively high incidence of local progression as recurrence after surgical removal. The quantitative assessment of TATE in the tumor stroma shows the statistically significant higher density of eosinophils in primary urothelial cancers tending to recur and no influence on DFS. In a similar study of colorectal cancer (in squamous cell cancer with different location), high grades of TATE were associated with the lesser tendency of tumor metastasis.Increased number of eosinophils may result from either an enhanced recruitment mediated by chemotactic mediators or a decreased apoptosis of eosinophils in the local tissue or both. The exact role of eosinophils in carcinogenesis - tumor transformation and/or progression is not quite clear.Mature human eosinophils contain numerous highly basic and cytotoxic granule proteins, such as major basic protein, eosinophil peroxidase, eosinophil cationic protein, and eosinophil-derived neurotoxin. Their activity is related to the release of these substances and basement membrane damage by eosinophil-derived cation proteins and peroxidase ,19-24. TTo the best of our knowledge, neither of these cellular interactions have been thoroughly studied in bladder cancer. Although most of the studies in other cancer location suggest a beneficial effect of TATE, the present results reveal reliable data for association of eosinophils in local recurrence of urothelial bladder cancer.Keeping in mind the different significance of TATE, in regard to their location – both intra- versus peritumoral and in different primary tumors, the most probable explanation seems to be that besides their direct interaction with tumor parenchymal and stromal cells, eosinophils have the potential to modulate the functions of other immune cells. Nevertheless, there is still a long way to understand the exact mechanism of the effect of eosinophils in cancer.At least in urothelial bladder cancer, it is tempting to suggest that TATE might be regarded as a possible prognostic factor for local recurrence. So far, the evaluation of eosinophils represents a promising and readily assessable tool, and should therefore routinely be commented on in the pathology report.The quantitative assessment of TATE in tumor stroma shows statistically significant higher TATE in primary urothelial carcinomas tending to recur. Although these findings need further explanation, it is tempting to suggest that TATE might be regarded as a possible prognostic factor for the local recurrence of bladder cancer.


The internal iliac artery is the main vascular supply of pelvic visceral structures. All pelvic surgeons must know the anatomic landmarks and basic steps of internal iliac artery ligation in order to stop massive pelvic hemorrhage. This cadaveric demonstration and clinical review of the internal iliac artery shows the anatomic landmarks and basic steps of internal iliac artery ligation. The anatomy of the internal iliac artery (IIA) has been well documented previously and it is the major blood supply of pelvic structures. It arises from the common iliac artery and runs infero-medially in the pelvis. An enormous number of small vessels, collateral circulation, and variations exist in pelvic vasculature ,2. The role of IIA ligation to control intractable pelvic hemorrhage  has been described by Kelly  for the Despite some technical difficulties with regard to anatomic relationships and potential complications,  ligation of IIA provides a rapid way to decrease the pelvic arterial blood flow. This clinical and photographic review shows the step-by-step surgical technique used in ligation of the IIA because all pelvic surgeons need to know how to ligate the IIA.The figures of this study were obtained during cadaveric dissections at the Consultants in Obstetrics and Gynecology-Management of Peripartum Bleeding and Morbidity Cadaveric Course, Bahçeşehir University Faculty of Medicine, İstanbul, 2017, and the Management of Peripartum Hemorrhage Cadaveric Course, Kafkas University Faculty of Medicine, Kars, 2018.Ligation of the IIA has a proven success rate in controlling massive pelvic hemorrhage, varying between 40% and 100%, and obstetric pathologies occupy the first place as the leading factor ,12. TablThe risk of operative injury beyond success is the major gap beneath the feasibility of the procedure . A detaiThe aorta is divided into left and right common iliac arteries at the level of the fourth and fifth lumbar vertebra (L4-5) and after a pathway of 4.0-5.0 cm the common iliac artery gives the branches of external iliac artery and IIA. The IIA runs infero-medially after the pelvic brim  and has Accurate identification of adjacent anatomic structures  will makMonopolar or bipolar electrocoagulation could also be used during dissection of the surgical field, bleeding from small veins will stop spontaneously; nevertheless, care must be taken. Before ligation of the IIA, dissection of the ureter is extremely important and inspection is better than just palpation of the ureter in the context of preventing any probable injury. Although ligation of IIA could be performed at any side of the patient either right or left, the surgeon must be careful during dissection and traction of the IIA because movement of the right-angle clamp from medial to lateral will cause a laceration on the external iliac vein.