The autonomic nervous system (cANS) is essential for proper heart function, and complications such as heart failure, arrhythmias and even sudden cardiac death are associated with an altered cANS function. A changed innervation state may underlie (part of) the atrial and ventricular arrhythmias observed after myocardial infarction. In other cardiac diseases, such as congenital heart disease, autonomic dysfunction may be related to disease outcome. This is also the case after heart transplantation, when the heart is denervated. Interest in the origin of the autonomic nerve system has renewed since the role of autonomic function in disease progression was recognized, and some plasticity in autonomic regeneration is evident. As with many pathological processes, autonomic dysfunction based on pathological innervation may be a partial recapitulation of the early development of innervation. As such, insight into the development of cardiac innervation and an understanding of the cellular background contributing to cardiac innervation during different phases of development is required. This review describes the development of the cANS and focuses on the cellular contributions, either directly by delivering cells or indirectly by secretion of necessary factors or cell-derivatives. The cardiac autonomic nervous system (cANS) modulates physiological cardiac functions such as heart rate and contraction force. The heart is innervated by nerves from the brain and spinal cord that either stimulate or inhibit these cardiac functions. Under pathological conditions, the density of these nerve fibers in the heart may be altered, leading to either too much activation (hyperinnervation) or too little (hypoinnervation). This  difference in innervation may cause a spatial imbalance in the activation of the heart, leading to cardiac autonomic dysfunction . CardiacThe development of innervation itself and the role it plays during the fetal period may be related to problems with innervation in later life. Pressure overload-induced hypertrophy is accompanied by regional hyperinnervation of sympathetic neurons and, interestingly, these neurons showed a fetal phenotype . TherefoThe development of cardiac autonomic innervation is a complex process that occurs in a temporally and spatially controlled manner. Many cell types are involved in establishing the cANS, either directly by delivering cellular building blocks, or indirectly by secreting factors that induce differentiation, repulsion, or attraction of other cells. This review focuses on cell populations that contribute to the development of the cANS, as well as on related neurotrophic factors and signaling pathways during embryonic and fetal development.The adult heart is a four-chambered muscular organ that maintains the pulmonary and systemic circulations. The cardiac muscle is self-exciting and has its own cardiac conduction system, consisting of specialized cardiomyocytes and conduction fibers which initiate rhythmic contractions. Development of the heart is conserved between vertebrate species  and, in To generate a third layer of the heart, the heart tube will be covered by an epicardial layer. Epicardial progenitor cells are found at E9.0  within the proepicardial organ (PEO), a cauliflower-like cluster of cells at the base of the inflow tract of the looping heart tube  18,19].,19.18,19During cardiac development, the peristaltic movement of the heart tube is replaced by a coordinated contraction, initiated by the sinoatrial (SA) node that passes the electrical activation through the other components of the cardiac conduction system. The initial immature base-to-apex activation pattern will develop into the mature apex-to-base activation pattern that is observed after ventricular septation .The early developing heart is not innervated yet, and the first signs of cardiac innervation during development are found at the dorsal mesocardium at E10.5 in mice . HoweverAlthough the development of cardiac innervation has been the subject of extensive investigations, the exact processes that govern normal cANS development are not known. Furthermore, the potential contributions of—and interplays between—various cell types are not well understood. In our attempt to link embryonic development to adult pathologies, differences between adult and embryonic cardiac function and anatomy are important. Given the complexity of the cANS, the paragraph below will address the anatomical structures of the cANS in the adult heart.The nervous system is anatomically divided in two main systems: the central nervous system (CNS) and the peripheral nervous system (PNS). The CNS includes brain and spinal cord, whereas the PNS contains cranial nerves, spinal nerves and ganglia. The PNS is again divided in a somatic (voluntary) and a visceral (vegetative) part, the latter including innervation of the heart. The visceral nervous system contains afferent (sensory) nerves that come from organs and vessels and project towards the CNS. In addition, the visceral system contains efferent (motoric) nerves to the heart, smooth muscle and glands; also known as the autonomic nervous system (ANS).Adult regulation of cardiac contraction and heart rate is achieved by innervation of the tissues of the cardiac conduction system through the cANS. Anatomically, the cANS is a complex system divided in sympathetic and parasympathetic elements A. All thSensory nerves have a different anatomy and function than the sympathetic and parasympathetic nerves. Their main cardiac function is to give feedback from baroreceptors on the aortic arch to the brain in order for the brain to react and maintain homeostasis. Sensory nerves provide information to the central nervous system and relay information on, for instance, carbon dioxide, blood pressure, and oxygen and sugar levels in the blood. The sensory neuronal body gives rise to axons which are sprouting to both heart and brain. The signal starts in the heart and is directed towards the brain via the neuronal body, in contrast to sympathetic and parasympathetic nerves where the neuronal body also indicates the starting site of the signal. The neuronal body of cardiac sensory nerve fibers lies either within dorsal root ganglia or nodose ganglia, depending on which nerve it accompanies. For instance, the afferent sensory elements that signal to the brain have their neuronal bodies in the nodose ganglion and they accompany the efferent parasympathetic elements of the (mixed) vagal nerve  innervation is still lacking, the chicken heart already responds to catecholaminergic stimulation ,41. The (Dbh) is disturbed, most homozygous mutant mice die in utero, likely due to cardiovascular failure; the surviving mutant mice, moreover, are smaller than their wild-type littermates. Only 5% of the homozygous mice reach adulthood . To. To67]. ing NCCs . Some mincreased ,74. Motincreased ,76. Thisncreased . The impdycardia ,78.+ neural crest cells in mice showed that neurotrophin receptor (NTR) p75-positive cells are already present in the dorsal mesocardium at E10.5, two days before the NCCs have reached that area. This alternative source of cells is thought to be from the nodose placode in the third pharyngeal arch, since parasympathetic ganglia are found to obtain cells and form cholinergic neurons derived from this area when the cardiac neural crest is ablated ,6430,64]The human adult heart contains around 700–900 cardiac parasympathetic ganglia, clustered together in subplexuses which are mostly found in the subepicardium of the atria . In the Subsequent sympathetic cardiac innervation is established in chick-quail chimeras from HH37–HH40, when TH-positive postganglionic fibers move alongside the previously developed vagal tracts to enter the heart through the cardiac plexus at the base of the heart  149,150150. ThesAfter entering the heart from the cardiac neural crest via the vagal tracts D, sympat+ veins recruit VSMCs originating from epicardial cells and/or NCCs . A. A151]. /or NCCs ,25,152. /or NCCs . At E15./or NCCs ,151. By Parasympathetic and sensory axon guidance are thought to follow slightly other paths than the sympathetic axon growth. There is evidence that sensory nerves do not follow vascular paths, but rather that arteries follow the path laid down by sensory nerves . AlthougParasympathetic axon guidance is probably induced by GDNF and its family members, but their role as guidance cues is unclear ,154. In Development of the autonomic nervous system (cANS) is a complex process involving many cell types, and is mainly determined by neural crest cell (NCC) delamination, migration and differentiation towards cardiac neurons. Other cell populations have a more supporting role by producing trophic factors to guide cells and axons (vascular smooth muscle cells (VSMCs) and endothelial cells), but also by secreting factors to induce NCC differentiation ) and survival . Further support may also come from delivery of cell-derivatives , sinus venosus and EPDCs) that help build guiding vessels  and fibroblasts), or that maintain cardiac function before and during development (intrinsic cardiac adrenergic (ICA) and small intensely fluorescent (SIF) cells). The contributions of the various cell types and their temporal distribution are summarized in Secreted factors control the patterning of nerves and in this review, several examples are given of factors involved in this process. For instance, SEMA3A is a neuronal chemorepellent and once affected, mice show disturbed innervation patterning with an imbalance in the epicardial-to-endocardial gradient. Interestingly, these mice exhibit sinus bradycardias and are highly susceptible to induced ventricular tachyarrhythmias , suggestWhether or not this increase of NGF is beneficial remains unclear. Some studies show that overexpression of NGF leads to hypertrophic sympathetic ganglia, hyperinnervation and cardiac dilation ,160. VenDuring chronic heart failure, the sympathetic nervous system is hyperactivated and the increased catecholamine levels ultimately result in detrimental effects on cardiomyocytes. This hyperactivation of the sympathetic nervous system also leads to increased levels of GRK2 and GRK5, related to desensitization of the adrenergic receptors ,165. ThiThe role of the sympathetic nerve system in pathology became even more elusive when researchers found that uninjured rats develop cardiac fibrosis and myocardial injury upon 6-hydroxydopamine-induced sympathetic denervation . When a Even though the cardiac development of several vertebrates is comparable, the postnatal change in heart rate during development may differ between mammalian species . The heaIn summary, much is still unclear regarding the development of cardiac innervation and the pathologies that are related to it. Further research is warranted to elucidate the process of pathological (hyper- or hypo-) innervation in order to improve quality of life in patients with heart disease and decrease the incidence of sudden cardiac death.


At the same time,the authors named the previous sepsis definitions Sepsis-1 (from 1991) and Sepsis-2 (from 2001) (On February 23om 2001) . The proSepsis became defined as a "life-threatening organ dysfunction caused by a dysregulatedhost response to infection."The method used to prepare the proposal was a retrospective analysis of large hospitaldatabases from two countries  in the attempt to establish the clinical and laboratoryparameters that best correlated with mortality among patients with suspectedinfection.To identify this cohort of patients with suspected infection in large hospital databases,the authors used non-validated criteria, including patients treated with antibioticswithin 72 hours after collection of biological samples for microbiological analysis orpatients subjected to sample collection up to 24 hours after the onset of antibiotictreatment. as thediagnostic criterion for sepsis. According to this suggestion, a patient with an acutechange in the SOFA score ≥ 2 meets the criteria for sepsis ]r sepsis . The tasHowever, due to the limitations of SOFA outside the intensive care unit (ICU), the taskforce recommended a new score . This instrument, which wasalso developed by the task force and was not validated in clinical practice, comprisedthree clinical parameters that were easy to assess  and wereIn turn, septic shock was defined as a "subset of sepsis in which underlying circulatoryand cellular metabolism abnormalities are profound enough to substantially increasemortality."The identification of patients with this condition followed another method and used theSurviving Sepsis Campaign database ;this method employed the Sepsis-2 definitions and clinical criteria for infection. Theexternal validation was based on data from two large American hospitals. The criteriafor septic shock became a cumulative presence of arterial hypotension (defined as theuse of vasopressors) and hyperlactatemia (> 18mg/dL or 2mmol/L) despite adequatevolume resuscitation .We emphasize that the category "severe sepsis" was eliminated, which according to theprevious criteria characterized septic patients with organ dysfunction andmanifestations of hypoperfusion or arterial hypotension associated with sepsis that inprognostic terms had a mortality rate intermediate between sepsis and septic shock.The medical community became divided over the clinical value of the new criteria. Thecriticism mainly focused on the following three aspects: (1) underlying theoreticalconcepts; (2) the methods used to define the criteria; and (3) their potentialimpacts on clinical practice.Regarding the theoretical aspects, the criticism emphasized the oddity of applyingdifferent criteria to the suspicion and identification of the same pathologicalphenomenon, which frequently exhibited the same clinical presentation, according towhether or not the patient was admitted to the ICU. The criticism stressed that thenew criteria stemmed from a purely retrospective analysis of hospital databasescreated for completely different purposes, were quite limited in their geographicdistribution, and defined for this particular objective, infection  as a "collection of biological samples + prescription of antibiotics withina given time interval"  and using physiological datacollected in a manner that was not completely explained . Clearly, this criticism only applies to the development of the criteria forsepsis and not to the criteria for septic shock, which as mentioned above are basedon another set of data.Without downplaying the relevance of the first two aspects, we believe that thefuture use of these criteria in clinical practice  is a cause of great concern. TheSepsis-3 criteria introduce no changes in the approach to sepsis, especiallyconcerning antibiotic treatment, fluid therapy, and vasopressor support, but neglectthe early identification of sepsis before the development of organ failure.-8Following the Sepsis-3 criteria, the previous categorizations of the severity andconsequent mortality due to infection that progressed from sepsis (infection meetingthe criteria for systemic inflammatory response syndrome or SIRS) to severe sepsis to septicshock  werereduced to simple infection, sepsis (infection and manifestations of organ failure),and septic shock  . Sepsis-The combination of the blood pressure values (or use of vasopressors after adequatevolume resuscitation) and lactatemia has been long known to allow the identificationof patients with different severities and prognoses, which to date were systematizedas follows:Cryptic shock: defined as lactate concentration ≥ 4 mmol/L withoutarterial hypotension (or use of vasopressors).Septic shock: hypotension induced by sepsis that persisted despiteadequate volume resuscitation and might present as:2.1. Vasoplegic shock: hypotension refractory to fluid therapywith normal serum lactate.2.2. Shock with tissue dysoxia: hypotension refractory to fluidtherapy with hyperlactatemia.The criteria defining the last group, which exhibits higher mortality, are thecriteria the authors of Sepsis-3 considered necessary for the definition of septicshock. In other words, the various phenotypic expressions of the severity of septicshock are not considered in Sepsis-3, because only dysoxic septic shock is takeninto account and the vasoplegic and cryptic shock categories are ignored. The lattercategories were classified as sepsis.Another major issue is whether we can undervalue the relevance of clinicalmanifestations that, according to the Task Force, are now called "infection" andthat until recently were septic patients with different morbidities and mortalities.Besides, the mortality of these conditions is not negligible, as may be inferredfrom the tables published by the authors of the Sepsis-3 manuscriptthemselves.As in every other situation, any change made should have a purpose. Are the previouscriteria less useful and restrictive for the management of more severe infections?The clinical evidence points to the opposite situation. The ultimate goal of ouraction as physicians is to reduce morbidity and mortality.The criteria for SIRS were the target of much criticism for having too highsensitivity but poor specificity. In turn, the term "severe sepsis", with itsconsequent organ dysfunction and/or tissue hypoperfusion and/or arterial hypotensionassociated with sepsis, was considered by several researchers  to be the true onset of septic conditions.From our perspective, the approach to sepsis should be grounded on three fundamentalaspects that should be considered simultaneously and based on demonstrated proof forits management and treatment as follows: (1) early recognition and stratification ofseverity; (2) prevention of and support for organ dysfunction based on an optimaloxygen delivery; and (3) treatment of the cause and control of the infectionsite. Admittedly, the Sepsis-2/SSC partnership has an optimal recordof success,,11,12 with asignificant impact on mortality by doing more with the available resources .To attain these goals, the Surviving Sepsis Campaign (SSC) sets of documents areavailable. These documents contain recommendations that indicate standardized,goal-oriented diagnostic and therapeutic actions according to the patient's severityand response to treatment based on early identification and stratification of sepsispatients (Sepsis-2). These recommendations were updated every four years, with thelatest version published in 2013.We should always keep in mind that there is no pathophysiological aspect that ispathognomonic of sepsis and that the diagnosis of infection results from theintersection of three vectors , because no specific marker is knownat present.In reality, we do not know whether the Sepsis-2 or the Sepsis-3 criteria bestidentify the most severe cases of infection that demand more timely therapeuticmanagement. However, we fear that downplaying infectious conditions that do not meetthe current Sepsis-3 criteria  will hinder their identification, resulting in an unnecessaryincrease in both morbidity and mortality due to their inexorable progression in thefollowing hours. We admit that this risk is purely theoretical at present.We anticipate that studies comparing the performance of both criteria in the realworld will be conducted in the near future. Independent from their results, ourapproach to the patient with suspected infection should always be clinical. Weshould strive to achieve the identification of the initial and sometimes subtlemanifestations of organ failure and hypoperfusion in all patients with suspectedinfection; however, these manifestations are devalued in the Sepsis-3 criteria infavor of scores (SOFA and qSOFA).,11-13 The authors of Sepsis-3 conclude their text by asserting,"These updated definitions and clinical criteria should clarify long-useddescriptors and facilitate earlier recognition and more timely management ofpatients with sepsis or at risk of developing it." Unfortunately, our perceptionsuggests the opposite outcome. SSC warns against this same risk by asserting, "Thefollowing advice is meant to put the recent publication of the consensus definitionsin context to facilitate the continued successes of sepsis screening, earlyidentification and treatment that have been the hallmark of SSC's qualityimprovement efforts associated with improved survival during the precedingdecade".Although not formally validated, the various criteria included in Sepsis-2 haveextraordinarily high sensitivity for the early stratification of infection. Whenthese criteria are followed by the application of the SSC recommendations, they havean impressive history of success in reducing the mortality of sepsis in severalareas of the world.


Trypanosoma brucei rhodesiense is associated with high mortality and is fatal if left untreated. Only a few studies have examined the psychological, social and economic impacts of rHAT. In this study, mixed qualitative and quantitative research methods were used to evaluate the socio-economic impacts of rHAT in Mambwe, Rufunsa, Mpika and Chama Districts of Zambia.Acute human African trypanosomiasis (rHAT) caused by Individuals diagnosed with rHAT from 2004 to 2014 were traced using hospital records and discussions with communities. Either they, or their families, were interviewed using a structured questionnaire and focus group discussions were conducted with affected communities. The burden of the disease was investigated using disability adjusted life years (DALYs), with and without discounting and age-weighting. The impact of long-term disabilities on the rHAT burden was also investigated.Sixty four cases were identified in the study. The majority were identified in second stage, and the mortality rate was high (12.5%). The total number of DALYs was 285 without discounting or age-weighting. When long-term disabilities were included this estimate increased by 50% to 462. The proportion of years lived with disability (YLD) increased from 6.4% to 37% of the undiscounted and un-age-weighted DALY total. When a more active surveillance method was applied in 2013–2014 the cases identified increased dramatically, suggesting a high level of under-reporting. Similarly, the proportion of females increased substantially, indicating that passive surveillance may be especially failing this group.An average of 4.9 months of productive time was lost per patient as a consequence of infection. The health consequences included pain, amnesia and physical disability. The social consequences included stigma, dropping out of education, loss of friends and self-esteem. Results obtained from focus group discussions revealed misconceptions among community members which could be attributed to lack of knowledge about rHAT.The social and economic impact of rHAT on rural households and communities is substantial. Improved surveillance and strengthening of local medical services are needed for early and accurate diagnosis. Disease prevention should be prioritised in communities at risk of rHAT, and interventions put in place to prevent zoonotic disease spill over from domestic animals and wildlife. Supportive measures to mitigate the long-term effects of disability due to rHAT are needed.The online version of this article (10.1186/s40249-017-0363-6) contains supplementary material, which is available to authorized users. Please see Additional file Trypanosoma brucei gambiense  and Trypanosoma brucei rhodosiense (eastern and southern Africa), spread by tsetse flies (Glossina ssp) . However the findings in our study did not account for under-reporting as described by Fèvre et al. . However, the latMany patients preferred to self-medicate and/or seek treatment from traditional healers until they became very ill, also contributing to late diagnoses. In agreement with Odiit et al. , the aveThis study also found that HAT cases and mortality rates were higher among males than females Table , with maAn important component of this study was the evaluation of the impact of long-term disabilities on the burden of rHAT. Long-term disabilities were common in this patient cohort and are not accounted for in standard HAT DALY calculations. The impact was significant with the number of DALYs increasing by between 44% and 50%, depending on whether age-weighting or discounting was applied or not  and results in loss of personal income and friends. This is in concordance with the findings of WHO  who repoThe lack of reliable data on the occurrence and outcome of infection for rHAT together with late presentation complicates treatment and disease prognosis. Moreover, limited awareness among community members and lack of prevention programmes result in a high disease burden and DALY for rHAT in rural communities. Poor diagnostic capacity of the local health system, specific therapeutic beliefs and practices, and the reluctance of patients to spend money on treatment until the illness becomes debilitating contributed to the high number of patients seeking health care in late stages of disease. As a neglected disease, analysis of the burden of rHAT on patient and households in specific geographical foci can help stimulate resource mobilisation for prevention and control programmes at a local and national level. Assessing the social and economic impacts of a neglected disease outbreak can help to contextualise the importance of prevention both to the affected population and the costs of outbreak mitigation strategies funded by governments and donors.rHAT is a disease that emerges out of poverty both in terms of transmission dynamics and clinical outcomes, while simultaneously perpetuating and reinforcing a state of poverty among affected households. The disease impacts households on multiple levels, including in areas of income generation, agriculture and food security, children’s education and long-term health consequences such as acquired muscular, nerve deformity and mental health (amnesia). At an individual level, the disease causes loss of self-esteem due to being stigmatised resulting from the acquired long-term physical and mental disabilities. Assessing the social and economic impacts of rHAT can help to contextualise the importance of prevention both to the affected population and the costs of outbreak mitigation strategies funded by governments and donors. An evaluation of the impact of long-term disabilities should be considered in future studies on the burden of rHAT.


The aim of this paper is to describe the use of a new ultrasound imagingprocessing technique to guide biopsies of suspicious breast calcifications. Weused this technique in 13 patients with suspicious breast calcifications thatcould not be submitted to stereotactic biopsy. Suspicious calcifications wereidentified by ultrasound, and the biopsy was successfully performed in allcases. Although mammography continues to be the method of choice for thedetection and characterization of microcalcifications, this new technique can bean alternative means of guiding biopsy procedures in selected patients who arenot candidates for stereotactic biopsy. However, despite the use ofalternative approaches,3, stereotactic biopsy is nottechnically achievable in all cases. Ultrasound has been proposed as an alternativeto stereotactic biopsy in selected patients with suspicious breastcalcifications,5. However, conventional gray-scaleexamination limits the identification of calcifications, especially those locatedoutside of a mass or duct, due to the lack of contrast with the normal breastparenchyma. Recent technical advances have improved the detection of calcificationsby ultrasound-8.Mammography is the method of choice to guide biopsies of suspicious breastcalcificationsThe aim of this paper is to describe the use of a new ultrasound imaging processingtechnique to identify and guide biopsies of suspicious breast calcificationsdetected on mammography.From June 2014 through June 2016, 13 patients with suspicious grouped breastcalcifications (< 2 cm in extent) that could not be submitted to stereotacticbiopsy were submitted to ultrasound-guided biopsy after the identification ofcalcifications using a new imaging processing technique . MicroPure is an image processing function designed toenhance the visualization of microcalcifications by ultrasound. It combinesnonlinear images and suppression techniques to highlight suspicious calcificationsas white spots on a blue background. During ultrasound examination, gray-scale andMicroPure images are displayed side-by-side on the screen. All of the patients gavewritten informed consent prior to the procedure. Three radiologists with experiencein breast imaging performed the ultrasound examinations and biopsies.Mammography images were analyzed to evaluate the distribution and topographiclocation of the calcifications. Ultrasound was then performed on an Aplio 500 system with a high-frequency linear transducer.After identification of the suspicious calcifications on ultrasound, 12-gauge coreneedle biopsy was performed. Specimen X-ray images were obtained to confirm that thecalcifications were present .In all 13 of the cases evaluated, stereotactic biopsy could not be performed, becauseof the location of the lesion in five cases ; because ofinsufficient thickness of the compressed breast in three; and because of clinicalconditions in five . Suspiciouscalcifications were identified using ultrasound, and the biopsy was successfullyperformed in all cases, without complications. The main technical difficulty was toidentify suspicious calcifications after removal of the first sample, due to thepresence of gas and edema/hematoma at the biopsy site. However, that did not impedethe removal of calcifications in the additional samples. Chest X-ray revealedmicrocalcifications in all specimens. ,8. Park et al. recently used the MicroPureultrasound function to guide biopsies of suspicious calcifications in nine patients.As in the present study, ultrasound biopsy was successfully performed in allpatients. Further large-scale studies are needed in order to assess the potentialcontribution of this new technique and to compare its performance with that ofstereotactic biopsy in the diagnosis of suspicious calcifications.The technique described here improves the capacity of ultrasound to detectcalcifications and increases the confidence of radiologists in their ability toperform ultrasound-guided biopsy in selected cases, when there are technicalchallenges for stereotactic biopsy due to the lesion location or clinicalcharacteristics of the patients. There have been few reports on the clinical utilityof the MicroPure technique in locating and evaluating breastcalcificationsIn conclusion, although mammography remains the gold standard for the histologicalevaluation of suspicious breast calcifications, ultrasound using innovative imagingprocessing techniques can be an alternative means of guiding biopsies, thus avoidingunnecessary surgery, in selected patients.


Today neurological diseases such as stroke represent one of the leading cause of long-term disability. Many research efforts have been focused on designing new and effective rehabilitation strategies. In particular, robotic treatment for upper limb stroke rehabilitation has received significant attention due to its ability to provide high-intensity and repetitive movement therapy with less effort than traditional methods. In addition, the development of non-invasive brain stimulation techniques such as transcranial Direct Current Stimulation (tDCS) has also demonstrated the capability of modulating brain excitability thus increasing motor performance. The combination of these two methods is expected to enhance functional and motor recovery after stroke; to this purpose, the current trends in this research field are presented and discussed through an in-depth analysis of the state-of-the-art. The heterogeneity and the restricted number of collected studies make difficult to perform a systematic review. However, the literature analysis of the published data seems to demonstrate that the association of tDCS with robotic training has the same clinical gain derived from robotic therapy alone. Future studies should investigate combined approach tailored to the individual patient's characteristics, critically evaluating the brain areas to be targeted and the induced functional changes. Stroke is one of the leading factors of morbidity and mortality worldwide  and human quality of life strongly depend on upper limb functioning  between two or multiple small electrodes on the scalp  loading effects in individuals with moderate to severe chronic stroke; however, it is insufficient to make significant changes at higher SABD loads  and tDCS has shown to be a potential neuromodulatory adjuvant of walking rehabilitation in patients with chronic stroke  to robotic locomotor exercises showed the capability to enhance the effectiveness of robotic gait training in chronic stroke patients  may be useful to improve the effects of robotic gait training in chronic stroke  the available results as well as discussing inconsistencies and possible issues. As a final goal, indications for the development of future and more specific rehabilitation protocols tailored to subject's needs are provided.The paper is structured as follows. In Section “Overview of the Main Studies on tDCS Coupled with Upper-Limb Robotic Treatment” an overview of clinical studies that analyze effects of tDCS combined with upper limb robotic therapy (RT) is reported.Section “Discussion” presents a critical discussion of the presented studies aimed to assess the efficacy of this novel combined approach. Finally, Section “Conclusions and future perspectives” reports final considerations and future suggestions.The study of the effects deriving from the coupled use of tDCS and RT represents a relatively young field of interest. In fact, the number of studies that have tried to investigate and prove the successful combination of these two techniques is limited.* OR ictus OR hemiplegia* AND robot* OR robotic therapy*, upper-limb rehabilitation, brain stimulation techniques, neurorehabilitation, rehabilitation robotics. Studies have been included only when focused on the novel therapeutic approach based on tDCS combined with robotic upper limb therapy.A wide literature search updated to January 2017 has been conducted resorting to the main databases, such as Pubmed Central (PMC), Cochrane, Scopus, Google Scholar. The following keywords have been employed: tDCS AND strokeBe a single session clinical trial  or controlled trial .Involve stroke patients.Concern movement therapy with a robotic device.Include transcranial Direct Current Stimulation (tDCS) as Non-Invasive Brain Stimulation Technique.Focus on upper-limb motor control .Use relevant motor control and functional ability outcome measures.Be a full-length publication in a peer-reviewed journal.The following inclusion criteria have been utilized:To enable the most complete overview of the current literature, the search has not been limited by patient subgroups  or by language.A flowchart of the search and inclusion process is shown in Figure The two conference papers Ang et al.  and MazzThe eight retrieved journal studies are Hesse et al. , Hesse eDescriptive features of the subjects;Protocol used within the study;Outcome measures of motor control, functional abilities and neurophysiological parameters of cortical excitability;Conclusions based on results.The following contents have been extracted from the analysis of the selected studies:p < 0.05) as calculated by an appropriate statistical tool.The results of the studies are considered positive if the difference between pre-post treatment and between robot-trained and control groups is significant , Modified Ashworth Scale (MAS), Barthel Index (BI), Medical Research Council (MRC) score for muscle strength, Box and Block test (B&B), Motor Activity Log (MAL), Action Research Arm Test (ARAT), Stroke Impact Scale (SIS)];Four neurophysiological parameters of motor cortical excitability ;Seven kinematic indices [Mean speed, peak speed, deviation, smoothness, duration, aim, and Hand Path Ratio  and cathodal tDCS is delivered on the contralateral hemisphere (inhibitory protocol). Sham stimulation is applied with an increasing current at training onset, and is switched off for the remainder of the stimulation time.Although the selected studies share the general objective of assessing the effects of tDCS combined with the RT, the employed investigation methods are different and provide heterogeneous data that are difficult to be analyzed in a systematic way. However, despite the difficulty to find a global primary outcome measure, interesting common features have been extracted and a list of factors has been identified.Effects of anodal and/or cathodal tDCS coupled with RT in post stroke patients ;Effects of different anodal tDCS delivering time, i.e., before, during or after RT;Effects of anodal tDCS compared to cathodal tDCS when coupled with RT;Effects of anodal tDCS combined with other neurostimulation techniques and RT.For the sake of clarity, the retrieved studies have been grouped in the following four categories while the main characteristics are reported in Tables The number of patients in the groups treated with real tDCS ranged from a minimum of 4  Table . On the The analysis of the literature presented in this paper has strengthened the efficacy of RT in stroke rehabilitation  or bilateral tDCS to different RT  did not produce significant effects in terms of FMS with respect to RT alone either in chronic or subacute stroke patients  in chronic stroke patients respect to MI-BCI treatment alone  and the lack of a standardized intervention protocol make difficult to compare and provide a definitive analysis of the results.However, an attempt of analyzing factors influencing outcomes herein presented has been carried out. Several factors have been identified and discussed as responsible for the large variety of the reported results. They are discussed in the following subsections.Type of intervention and treatment intensity may represent crucial aspects for evaluating efficacy of tDCS coupled with robot-aided rehabilitation. In fact, from the literature analysis emerges that tDCS and RT effects have been investigated in multiple manners, thus producing different results.For instance, the feasibility of the combined approach (tDCS and RT) both in chronic and in subacute stroke patients  might have influenced the current findings.Another important factor to be considered is the delivering time of tDCS respect to RT as well as the inhibitory and/or excitatory approach. In fact, duration, frequency, timing, and polarization of the stimulation are different in every study.Furthermore, the use of different techniques such as PNS   it might be preferable to use the excitatory approach rather the inhibitory.Recent findings suggest a possible alternative approach of neuromodulation. This method is based on the inhibition of the ipsilesional motor cortex aimed to favor motor learning through mechanisms of “homeostatic” plasticity  with upper limb robotic therapy. The large variability in the characteristics of patients enrolled in different studies  and the lack of a standardized intervention protocol make difficult to reach a definite conclusion.Restoration of motor control seems to be the same after electrical brain stimulation coupled with RT and after RT alone as supported by a quantitative analysis of FMSs.Furthermore, these findings confirm the effectiveness of RT in post-stroke patients.However, slight encouraging effects arise accounting for the lesion site, the type of stroke and type of damage thus suggesting the development of patient-tailored rehabilitative treatments.In future studies, a more individually tailored approach is required to choose the best type of tDCS (excitatory and/or inhibitory) and the best possible target  after taking into consideration several parameters. Moreover, such studies will help to refine the bimodal balance–recovery model and test whether the efficacy of individualized NIBS therapy combined with robotic therapy might result superior to the current methods.DS and LZ designed the paper, analyzed the literature, analyzed robot data, and wrote the manuscript. StM partly analyzed the clinical literature together with SaM, MB, and GM and they also revised the manuscripts. FR revised the manuscript and contributed to write clinical parts related to neuromodulation. EG, VD, and SS supervised the writing. All the authors read and approved the final version of 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. The reviewer PL and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


The most frequently used method of analysis of trace organic pollutants in ambient air is gas chromatography-mass spectrometry (GC-MS). Presently, target compounds in air monitoring samples are identified from gas chromatographic retention times and a combined forward-reverse search of mass spectral reference spectra. The primary objective of this study is to develop computational pattern recognition procedures for the identification of chemical classes and, if possible, individual compounds from routine GC-MS data files. The additional information on nontarget compounds would supplement compound identifications obtained from the target list. The target set of compounds investigated consisted of 78 substituted benzenes, haloalkanes and haloalkenes.SIMCA pattern recognition was developed by Wold and coworkers  for applIn this study an IBM PC-XT microcomputer with 640K memory was used. A modified version of the SIMCA program was used for the majority of the pattern recognition calculations. The low resolution mass spectra of 78 compounds were obtained from the EPA-NIH Mass Spectral Library on an INCOS data system see . The GC-The data were preprocessed by taking the autocorrelation transform of the mass spectra for mass lags less than 100. In the initial stages of class modeling, the autocorrelation transformed spectra for the 78 training compounds were examined for class separation with two and three dimensional principal components projections of the training data. Three different groups were found: nonhalogenated benzenes; chloroaromatics, chloroalkanes, chloroalkenes; and bromocarbons.Principal components models were then derived for each class . None ofOnce a class assignment is made, further identification of each spectrum can be made. This is done by calculating the distance, using the entire spectrum in autocorrelation space, of each compound to its three nearest neighbors in its assigned class. To obtain a specific structural assignment, the normalized correlation coefficient of the unknown mass spectrum with that of each of its three nearest neighbors is obtained. If the unknown is not identified as one of the three nearest neighbors in the first class assignment, the correlation coefficients of the three nearest neighbors in the second SIMCA class assignment are compared.Three direct injection calibration samples, which contained all but one of the training set compounds, were analyzed . Of the The GC-MS data files for three field samples were also analyzed . The ideThere were 75 different nontarget compounds identified in 120 occurrences in the three samples. The classification results agreed very well for the two class 1 and class 2 spectra. However, a very large number of alkanes and alkenes were incorrectly classified as chlorocompounds. Further details of this study are given in references , and 4]4].Although the research described in this article has been funded by the U.S. Environmental Protection Agency under Cooperative Agreement CR-811617 with the University of Illinois at Chicago, it has not been subjected to Agency review. The mention of commercial products does not constitute endorsement or recommendation for use.


This is a review of the book Spectral Domain Optical Coherence Tomography in Macular Diseases edited by Meyer et al. . The bo The outstanding book Spectral Domain Optical Coherence Tomography in Macular Diseases edited by Carsten et al. . OpticalThe book Spectral Domain Optical Coherence Tomography in Macular Diseases was produced by three brilliant Editors and over 60 Chapters authors. This 1st Edition by Springer Publisher. By gathering some of the most famous experts in the field to contribute chapters, Profs. Meyer, Saxena and Sadda were able to offer a superb work with several key features including point-to-point correlation of retinal conditions and spectral-domain OCT, 3-dimentional evaluation and segmentation analysis and the surgical use of OCT in vitreoretinal diseases management. The result is a streamlined, efficient teaching book that should be enjoyed as reference text for clinical practice in retina.The book covers OCT in detail and comprehensively in five parts throughout the more the 400 pages. In the first part, the readers may appreciate a detailed review of all technologies available on OCT including en-face, OCT-angiography, and swept-source OCT. Worthy to highlight is the excellent introductory chapter by Sisha et al. named OCT: a Primer. In the second chapter, the role of OCT for the most common retinal diseases are presented. The OCT images are of high quality and well labeled. The focus was not only the diagnostic importance of OCT test, but also its importance in clinical decision making. The third part reviews the use of OCT in diseases caused by anomalous vitreous adherence, whereas the fourth covers few interesting OCT investigations in not so common but also important retinal conditions. Finally, the last fifth part summarized and provided future directions to the revolutionary OCT technology.The new book Spectral Domain Optical Coherence Tomography in Macular Diseases provided clinicians both basic and advanced understanding of OCT applications in the medical field. The book will be useful for residents and fellows interested in pursuing an education in the retina medical specialty as well as practicing ophthalmologists. Overall, it has included the multiple applications and interpretation of OCT related to the posterior segment of the eye. Advances in OCT technology provided better understanding of pathogenesis, enhanced monitoring of progression and evaluation of response to therapy in diseases of the retina. Further improvements in either software or hardware should further advance the clinician’s ability to assess and manage chorioretinal diseases. We strongly suggest the book to all colleagues interested in res clinical applications of OCT in ophthalmology and retina.


Like other previous treatments and approaches, sorafenib, an antiangiogenic drug, failed to show any benefit in the adjuvant setting for hepatocellular carcinoma in a large clinical trial. We discuss reasons and implications of these negative results and the implications for clinical practice and future research.NCT00692770. Registered 5 June 2008. This study has been completed.ClinicalTrials.gov:  Lancet Oncology by Bruix and colleagues [The prevention and the delay of relapse of hepatocellular carcinoma (HCC) after curative treatment remain a major challenge. Recently the sorafenib as adjuvant treatment in the prevention of recurrence of hepatocellular carcinoma (STORM) study reported in lleagues  investigFirst, it has important implications for the design of future adjuvant trials in HCC and for the choice of the drug to be tested. The goal of adjuvant treatment is to reach for a cure. When tumors are emerging from a pro-tumoral underlying disease, decreasing the risk of both recurrence and de novo tumors is an additional challenge. Whether these two goals are achievable with a unique drug and strategy is an open question in HCC. Assuming side effects for a limited period of time, most active treatments should effectively target tumor residues. In contrast, long-term preventive treatment of the underlying disease would require drugs with a very good tolerance profile fitting chronic administration. The latter is a challenge in the drug development paradigm in which molecules are developed for maximum efficacy in the advanced setting. However, as illustrated by this trial and others, these results are quite often not translatable to the adjuvant setting. Undetectable tumors may exist as vascularized micrometastases, non-vascularized clusters of tumor cells, and isolated or even circulating tumor cells . WhereasIn the STORM trial, no beneficial or detrimental effect of sorafenib has been reported besides a trend towards increased time to recurrence, which may reflect a positive impact of sorafenib on tumor relapse and/or prevention of de novo tumors. Investigating post-recurrence treatment strategies and outcomes may also yield valuable information in the perspective of global HCC management.Second, adjuvant strategy in HCC crucially needs improved stratification to better define the population who could benefit from adjuvant treatment from the population who does not need further therapeutic intervention. Considering the results of the present study and the preceding considerations, new trials directed to patients with a higher risk of relapse than the STORM population might be worthwhile to redeem the effect of sorafenib as adjuvant treatment. Nevertheless, improved staging combining histological and biological parameters would be an important step forward discriminating subgroups prognosis and therapeutic interventions.Finally, in the present time of exploding medical costs, adjuvant trials should consider the cost-effectiveness of long-term treatments, which emphasize the need of better stratifying patients to focus on the appropriate population . As hypo


Selenium is known to possess both genotoxic and antigenotoxic properties. In the present study, we have evaluated the genotoxicity and antigenotoxicity of three selenium compounds  by measuring in vitro micronucleus induction. Assays were conducted in whole blood lymphocytes and in the TK6 lymphoblastoid cell line, with and without co-treatment with potassium dichromate, a known genotoxic compound. In general, the compounds were more active in TK6 cells than they were in blood lymphocytes. Only 1 μM selenous acid increased the frequency of binucleated cells containing micronuclei (BNMN) in blood lymphocytes, while all three selenium compounds increased BNMN in TK6 cells. In addition, combinations of selenous acid and potassium dichromate resulted in lower frequencies of BNMN than potassium dichromate alone in blood lymphocytes, while combinations of sodium selenate and potassium dichromate produced lower frequencies of BNMN than potassium dichromate alone in TK6 cells. The concentrations of selenium compounds that were used, in combination with the medium components and the biological physiology of the whole blood lymphocytes and TK6 cells, could have affected the redox potential of the compounds, switching the chemicals from a pro-oxidant to antioxidant status and vice-versa. The lower activities of the compounds in blood lymphocytes may be due to the protective effects of blood components. The results indicate that the genotoxic and antigenotoxic properties of selenium compounds are highly dependent upon the conditions under which they are evaluated.


Fusiform aneurysms in the posterior inferior cerebellar artery (PICA) are rare and challenging to treat. Surgical treatment options for a fusiform aneurysm in the PICA include trapping with/without bypass and wrap-clipping, when elimination of the pathological wall from the systemic circulation and prevention of perforator injury are important. In addition, lower cranial nerve impairment due to surgical manipulation should also be avoided.A fusiform-shaped aneurysm was found in a proximal part of the PICA by magnetic resonance angiography undertaken for evaluation of repeated vertigo in a 36-year-old man. The patient underwent direct surgery via a lateral suboccipital transcondylar fossa approach. The entrance of the pseudolumen was the only part to be wrapped and obstructed by clip application, through the corridor between the acoustic and glossopharyngeal nerves to avoid lower cranial nerve injury.Indocyanine green (ICG) videoangiography demonstrated obliteration of pseudolumen and patency of peripheral PICA and perforator contributing to the medulla oblongata. The postoperative course was uneventful without periprocedural complications, including dysphagia and hoarseness.Partial wrap-clipping technique for obstruction of the entrance into a pseudolumen is one of alternatives for dissecting fusiform-shaped aneurysm in the PICA. ICG videoangiography was helpful to confirm the obliteration of the pseudolumen and patency of parent vessel and perforators. Aneurysms located in the peripheral posterior inferior cerebellar artery (PICA) are rare and account for less than 1% of all intracranial aneurysms . FusiforHerein we present the case of a dissecting fusiform aneurysm in the PICA with tendency for growing. In this case, only the entrance of the pseudolumen separated by the intimal flap was obstructed with the partial wrap-clipping technique in order to avoid lower cranial nerve injury. Indocyanine green (ICG) videoangiography was helpful to confirm the obliteration of the pseudolumen and patency of the parent vessel and perforators.A 36-year-old man presented with a 1-year history of repeated vertigo, when magnetic resonance angiography (MRA) demonstrated an aneurysm originating from the proximal part of right PICA. Follow-up MRA showed an increase in size of the aneurysm after 6 months. Besides MRA, digital subtraction angiography and three-dimensional computed tomographic angiography (3DCTA) indicated that the aneurysm appeared to have an ellipsoid shape with an 8-mm maximum diameter and a distinct neck, and was diagnosed as a congenital berry aneurysm Fig. , which wProximal ligation, trapping, and resection have been reported as curative treatment options for intracranial arterial dissecting or fusiform aneurysms, as well as endovascular internal trapping. Even though these treatments are reasonable for the concept of elimination of pathological vascular walls from the systemic flow, normal blood flow would be consequently sacrificed to require vascular reconstruction. Even if that were the case, the danger of perforating arterial injury would remain.The wrap-clipping method has been established as one of treatment options for arterial dissection and blood blister aneurysm (BBA), which can provide reinforcement of holding the applied clip to result in prevention of re-rupture and enlargement of the pathological wall –3. This A variety of materials have been reported for wrapping, including muscle fascia, cotton, Goatex, and Dacron , 3. For A dissecting aneurysm occasionally has a fusiform shape besides a tapered occlusion and pearl and string signs. On cerebral angiography, some fusiform aneurysms show the intimal flap itself, like in our case, as well as a double lumen, pooling, and/or slow filling along the vessel, as an evidence of dissection. However, many of them do not reveal any apparent findings supporting a dissecting nature. For precise diagnosis of fusiform aneurysm, not only catheter angiography and 3DCTA that mainly provide information regarding the intra-vascular space but also MRI are important for depicting the character of the wall and thrombus inside the aneurysm. Basi-parallel anatomical scanning (BPAS) MRI has been reported to be useful for diagnosis of vertebral artery dissection . MRI-BPAIn this paper, only the entrance of the pseudolumen was obliterated with partial clip application Fig. . MeticulThe wrap-clip technique for obliteration of only the entrance into a pseudolumen might be one of the alternatives for treating a dissecting fusiform-shaped aneurysm, which was effective for avoidance of lower cranial nerve injury in the case of the proximal part of the PICA. ICG videoangiography played an extremely important role in confirmation of obliteration of the pseudolumen and patency of the parent vessel and perforators.


Saccharomyces cerevisiae that serves until today as an important model organism for septin research. In yeast, the septins assemble into a highly ordered array of filaments at the mother bud neck. The septins are regulators of spatial compartmentalization in yeast and act as key players in cytokinesis. This minireview summarizes the recent findings about structural features and cell biology of the yeast septins.The septins are a conserved family of GTP-binding proteins present in all eukaryotic cells except plants. They were originally discovered in the baker's yeast  Saccharomyces cerevisiae . These were later classified as members of the septins, a new family of cytoskeletal proteins in the budding yeast , was identified more than two decades after the initial screen , only one structure from a yeast septin was solved (Cdc11). Structure determination, the evaluation of the role of posttranslational modifications of the septins and the uncovering of so far unknown septin related processes in the living cell will represent the challenges for septin biologists for the next decade.Recapitulatory, the past few years have witnessed a considerable increase of published results concerning yeast septins, the majority of these in budding yeast. Efforts were made in all areas of septin biology, from structural investigations using EM, light microscopy and X-ray diffraction to reconstruction of septin assembly in complex TG and OG wrote the manuscript. OG prepared the figures.OG was supported by Deutsche Forschungsgemeinschaft grant JO 187/8-1.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.


Multifunctional nanoparticle systems can integrate imaging, targeting and treatment moieties on the surface and in the core, resulting in targeted delivery of the imaging or treatment modalities, specifically to the tumor. Multifunctional nanoparticles also enable simultaneous delivery of multiple treatment agents, resulting in effective combinatorial therapeutic regimens against cancer. In this review, various multifunctional nanoparticle systems that feature a variety of targeting moieties for  Multifunctional nanoparticle-based platforms of anti-cancer drug delivery have paved the way for innovative therapies that are more efficacious, less invasive and less toxic. Nanoparticle carriers provide significant advantages over conventional therapy modalities . NanoparA significant advantage of using nanosized carriers is their ability to be targeted actively to the tumor sites. Conventional chemotherapy drugs are cytotoxic and kill actively dividing cancer cells, but can also affect other healthy dividing cells. The surface area/volume ratio and the chemistry of NPs afford the attachment of cancer-specific molecules on the NP surface that can specifically bind to their targets on the cancer cell. Internalization of the NP into the cell results in higher cellular uptake of the drug/active agent and higher anti-tumor activity. Targeted delivery isolates the effect of the drug to just the cancer cells expressing the targeted molecule, decreasing the systemic toxicity and the side effects of the drug or active agent, some of which could be life threatening .In contrast to monofunctional NPs that deliver only a single payload of drug or active agent, multifunctional NPs can integrate various functionalities inside the core of the NP or on the NP surface to synergistically achieve maximal anti-tumoral activity . It has Multifunctional NPs can also be loaded with imaging agents or molecules to provide diagnostic information during optical imaging, magnetic resonance and photothermal detection . OverallA number of organic and inorganic materials have been used to fabricate multifunctional NPs with their own distinctive architecture and attached functionalities , and thel-lysine, aspartic acid, d,l-lactic acid and spermine, while the outer corona consists of hydrophilic blocks, such as poly(ethylene glycol) (PEG), poly(N-vinyl-2-pyrrolidone) (PVP) and poly  sebacate, or P ovarian carcinoma cells and demonstrated the synergistic and sustained delivery of both agents [A comprehensive understanding of the chemistry and the pharmacokinetics of these various drugs and other biologic agents that have been utilized in the treatment of multiple cancers is imperative for the construction of sophisticated multifunctional drug platforms. The combination of hydrophilic  or hydrophobic  drugs with negatively-charged siRNA or DNA can target different metabolic pathways of tumor cells, resulting in a multipronged attack on the cells . In a brst Bcl-2 . Both ofh agents . Similarh agents .et al. developed a poly(ethylene glycol)-dioleoylphosphatidyl ethanolamine (PEG-DOPE)-modified G(4)-PAMAM nanocarrier that could deliver a drug-siRNA combination payload to tumor cells. The lipid modification of cationic polymers increased the transfection efficiency, and the micellar dendrimer system yielded higher stability and protection of the siRNA against enzymatic degradation [Several nanoformulations of siRNA and shRNA inhibitory sequences against genes that suppress the expression of the various functional and metabolic pathways of cancer cells have been developed and successfully used to inhibit the growth and progression of cancer. Biswas radation . Paclitaradation ,36, resuradation .The next section discusses the other arm of an efficient anti-cancer multifunctional NP: the targeting moiety that can specifically recognize tumor cells and maximally deliver drug/active agent payloads to the tumor site.One of the major advantages of nanotherapy in cancer is its capability to distinguish between cancerous and healthy tissue and, therefore, to minimize the systemic toxicity associated with chemotherapy. The size and pharmacokinetic properties of the NPs ensure that they accumulate at tumor sites. This is done through passive targeting, which is the preferential accumulation of NPs at tumor sites and is dependent on the pathophysiology of the tumor mass as a whole and the uniqueness of the tumor microenvironment. The irregular and porous blood vessel network surrounding the tumor allows the penetration of NP drug delivery complexes in a size-dependent manner. Passive delivery can be further enhanced by modification of the nanomaterials’ surface using different hydrophilic spacers, such as poloxamer, polyethylene oxide (PEO), PEG, lauryl ethers and polysorbate .l-arginine glycine l-aspartic acid (RGD)) and oligonucleotides (aptamers) have been used as ligands to recognize target receptors on the tumor cells -encapsulated magnetic NPs have been used to detect neuroblastoma cancer cells via MRI along with subcellular fluorescence imaging. Magnetic NPs have also been coupled to radionuclides, such as 124I, to engineer MRI-PET probes that can accurately detect lymph node metastasis in patients [Magnetic NPs have been used as the platform on which other functional molecules, such as fluorescent probes or radionuclides, can be attached to form dual modal imaging probes. Fluorescent dye-doped silica (DySiOpatients .64Cu radiolabeled iron oxide NP conjugated to the tumor binding RGD peptide was used for simultaneous MRI and positron emission tomography (PET) imaging of neuroblastoma tumors [Hybrid modalities that have multiple imaging probes engineered into one NP structure synergistically combine the advantages offered by each specific imaging technique that can lead to earlier and more efficient detection of tumors and can also be used for real-time monitoring of therapeutic efficiency. A hybrid NP consisting of an iron oxide NP coated with a PEG-grafted chitosan polymer and conjugated to fluorescent Cy5.5 and a tumor targeting moiety was found to accumulate in medulloblastoma tumor tissue in transgenic mice. The signal intensity increased over the first 50 h after an intravenous injection and stayed steady for the next 70 h . Similara tumors . As discQuantum dots have also been successfully employed in drug or gene delivery and imaging. A quantum dot-aptamer-doxorubicin (QD-Apt-DOX) conjugate was constructed by surface modification and attachment of the prostate tumor cell-specific A10RNA aptamer to the CdSe-ZnS core of the quantum dot that also was intercalated with doxorubicin. Upon internalization and release of doxorubicin, optical activation techniques can detect the fluorescence associated with this nanocomplex and can spatially locate the tumor, visualize drug delivery and uptake and detect the real-time response of the tumor cell to the drug .Gold NPs have also been widely used in the fabrication of cancer-targeting multimodal drug delivery systems, because of the ease of synthesis, the presence of negative reactive groups on the surface that can be easily modified for conjugation and low toxicity . A multiThe development of NPs that can serve as platforms for multicolor optical imaging provides the opportunity to obtain information about multiple molecular parameters associated with the cellular and metabolic processes of the tumor cell . The useTargeted and combinatorial NP therapies are therefore the future of cancer therapeutics and personalized medicine. Advances in genomics and proteomics have resulted in increasing information about the molecular profiles and biomarkers of various cancers. This information will aid in the fabrication of novel multifunctional NPs that can target tumor cells with more precision and specificity. Newer and more sophisticated multimodal imaging capabilities ensure early disease diagnosis, real-time monitoring and detection of its response to the treatment regimen. The power of nanotechnology in integrating all of these functionalities has caused a paradigm shift in the way cancer is treated and is inarguably the future of cancer therapy and cancer patient care.


A new scanning probe microscopy images the nanoscale world with synergistic force and infrared light at 10-nm spatial resolution. 3NH3PbBr3 perovskite crystals are studied with correlative infrared/mechanical nanoimaging. Furthermore, we show that the PFIR method is sensitive to the presence of surface phonon polaritons in boron nitride nanotubes. PFIR microscopy will provide a powerful analytical tool for explorations at the nanoscale across wide disciplines.Nondestructive chemical and mechanical measurements of materials with ~10-nm spatial resolution together with topography provide rich information on the compositions and organizations of heterogeneous materials and nanoscale objects. However, multimodal nanoscale correlations are difficult to achieve because of the limitation on spatial resolution of optical microscopy and constraints from instrumental complexities. We report a novel noninvasive spectroscopic scanning probe microscopy method—peak force infrared (PFIR) microscopy—that allows chemical imaging, collection of broadband infrared spectra, and mechanical mapping at a spatial resolution of 10 nm. In our technique, chemical absorption information is directly encoded in the withdraw curve of the peak force tapping cycle after illumination with synchronized infrared laser pulses in a simple apparatus. Nanoscale phase separation in block copolymers and inhomogeneity in CH Properties of materials depend on the composition and organization of their constituent components at the small scale. In a variety of disciplines of nanoscience and nanotechnology, characterizations of the chemical compositions, mechanical properties, and geometric shapes with tens of nanometers of spatial resolution are an essential part of everyday research. A nondestructive, ambient-compatible, spectromicroscopy method that can deliver chemical-sensitive imaging, the broadband spectrum from the nanoscale, together with the topographic and mechanical properties with a spatial resolution of ~10 nm, is highly desirable for the broad research communities. These joint characterization abilities would undoubtedly accelerate the research across a broad range of research fields. However, current analytical techniques are insufficient to provide the desired multimodal characterization abilities.The material response to infrared light is indicative of its chemical composition because infrared absorptions directly correspond to the vibrational modes of functional groups in chemical materials and lattice vibrations in crystals. However, the spatial resolution of conventional Fourier transform infrared (FTIR) microscopy is bound by the optical diffraction limit to half of the wavelength , uses a sharp metal-coated probe position-controlled by an atomic force microscope (AFM) to locally enhance the optical field and to excite the vibrational  (A recent innovation of infrared nanoscopy is photoinduced force microscopy (PiFM) through dipole-dipole interactions between the metallic probe and the induced dipoles of the sample  microscopy. It enables spatial imaging of infrared absorption responses with a spatial resolution of ~10 nm and the collection of broadband infrared spectra from nanoscale locations, together with simultaneous nanoscale mapping of mechanical properties of modulus and adhesion. The method is an action-based spectromicroscopy technique that combines synchronized infrared pulse excitations with the time-gated detection of the laser-induced mechanical responses in the sample. It operates in the peak force tapping mode of AFM (−1. The contact resonance oscillations and the baseline offset (defined as Δ in −1. Far-field FTIR spectrum from the PTFE bulk is displayed (black dashed curve) as a reference, with which the PFIR spectra show excellent agreement. In −1 (a similar plot displaying the pulse energy versus the baseline offset is given in fig. S2). A linear energy dependence is observed, which confirms that the laser-induced mechanical effect is proportional to the incident laser power. Note that the behavior of laser-induced mechanical responses is sample-dependent. The PFIR characteristics of poly(methyl methacrylate)  exhibit a more complex energy dependence than those of PTFE, which indicates the presence of a glass transition in the high–laser power regime. The practical implication of the power dependence is that the energy of the pulsed laser source should be kept low within the linear regime to avoid the glass transition when studying polymers.The schematics of the PFIR apparatus are shown in −1 reveals that the vibrational resonance of P2VP leads to enhanced PFIR signal. In contrast, the substrate lacks infrared resonances, resulting in a small background response. The maps of elastic modulus and adhesion extracted from the Derjaguin-Muller-Toporov (DMT) model  vibrational modes are enough to generate a detectable PFIR signal with a signal-to-noise ratio of 3. This observation suggests that PFIR is a highly sensitive infrared imaging technique. The high signal sensitivity is likely partially due to the presence of enhancement of the infrared field from the gold substrate as well as from the platinum-coated probe. The gap formed between the gold surface and the metallic probe can locally enhance the electromagnetic field to improve the signal  because it is not accessible by direct far-field infrared radiation. −1. Nonuniform distributions of the infrared absorption responses are revealed by the PFIR images. For comparison, s-SNOM images taken at the same frequencies are given for the same nanotube in −1, the whole segment of the BNNT shows both s-SNOM and PFIR responses. At 1430 cm−1, only a part of the segment shows the s-SNOM and PFIR responses. Detailed examination of the PFIR images reveals the edges of twisting polygonal facets of the BNNT, which were known in the literature (The boron nitride nanotube (BNNT) is known to support surface phonon polaritons (SPhPs) in the spectral region of negative dielectric constants above a strong phonon resonance  spectral region. The spectra from different locations exhibit variations of the PFIR spectra. For data taken close to the terminal of the BNNT , the high-frequency (>1420 cm−1) SPhP mode appears in the PFIR spectra. At locations far from the terminal, the high-frequency SPhP mode gradually disappears. This observation can be explained by the fact that high-frequency SPhP decays faster and, therefore, is less detectable when the probe is far from the terminal ; but a low set point reduces the amplitude readout from the contact resonance, because the cantilever is not efficiently anchored on the surface with a low peak force set point. A peak force set point of 4 nN was found to be good for the measurement using the MikroMasch HQ:NSC14/Pt probe with a spring constant of 5 N/m. In the point spectroscopy mode of PFIR microscopy, the probe of the AFM stayed at a fixed location while maintaining the peak force tapping feedback. The frequency of the QCL was scanned. At each frequency, the PFIR signals were recorded to form the PFIR spectra that are proportional to the infrared absorption spectra. The frequencies of the QCL were tuned across the spectral range while avoiding strong water absorption lines in the infrared fingerprint region. The power of the laser at different wavelengths was measured with a mercury cadmium telluride (MCT) detector  or with reference nonresonant samples and used to normalize the amplitude-collected PFIR spectra to compensate for residual water absorption or nonuniform power over the QCL tuning range.The mechanical measurement was performed with the peak force tapping simultaneously with the PFIR signal extraction. The real-time indentation of the sample was averaged, analyzed, and fitted with the DMT model  was dissolved in toluene and spin-coated on a gold substrate. The spin-coater (KW-4A) uses 620 revolutions per minute (rpm) for 9 s and 2560 rpm for 20 s. The thickness of the block copolymer was found to be 30 nm. The perovskite sample was synthesized according to the procedures described in the study by Yang et al.  was prepared by drop-casting the solution of P2VP in toluene on a gold substrate. The PS-


MicroRNAs play important roles in the development of human cancers. This case-control study is to evaluate the roles of the polymorphisms in pre-miRNAs on risk of oral cancer in a Chinese population.The genotypes of three polymorphisms were determined in 340 patients with oral squamous cell cancer and 340 healthy controls who were frequency matched for age and sex. Odds ratios (ORs) and 95% confidence intervals (95%CIs) were calculated to assess the association. All analyses were performed using the SPSS software. 3.154 0.001.For miR-499 rs3746444, individuals carrying homozygous CC genotype had increased risks of oral cancer compared with the homozygous wild TT genotype . The C allele of miR-499 rs3746444 was associated with a higher risk of oral cancer with significant odds ratio of 1.453. In the stratified analyses by sex, the associations between miR-499 rs3746444 and miR-146a rs2910164 polymorphisms with the susceptibility of oral squamous cell cancer were significant in males. However, with 1/4 as many subjects there were no significant associations between the three polymorphisms and oral cancer risks in females. The joint effects of miRNA polymorphisms and smoking on the risk of OSCC were analyzed and the results suggested that the association between microRNA genetic variants and OSCC risk was modified by smoking.These findings suggest that miR-499 rs3746444 and miR-146a rs2910164 polymorphisms may contribute to genetic susceptibility to oral squamous cell cancer. It is estimated that there are 263,900 new cases and 128,000 deaths from oral cavity cancer in 2008 worldwide [Molecular epidemiologic studies showed that there were hundreds of genes involved in OSCC , in whicThe most widely studied genetic alterations in recent years are single nucleotide polymorphisms (SNPs). SNPs within the seed region or in the precursor stem-loop of miRNA may significantly affect the production or processing of miRNA –14. Now This is a hospital-based case-control study of oral cancer in Shenyang City, located in northeast China. The case group consisted of 340 pathologically confirmed diagnosed OSCC patients (between January 2012 and September 2014) at Hospital of Stomatology, China Medical University. During the same period, 340 controls were recruited from other patients with no evidence or history of cancer in the same institute. Controls were frequency matched to cases on age (±5 years) and same sex. Participants were unrelated ethnic Han Chinese. Each participant contributed 10ml venous blood and was interviewed to collect related data at the time they were admitted into the study. The written informed consent was obtained from each participant, and human investigations were approved by the Institutional Review Board of Hospital of Stomatology, China Medical University.Genomic DNA samples of all cases and controls were isolated by Phenol-chloroform Method. Genotyping of the miRNA SNPs was done using Taqman allelic discrimination method  on an Applied Biosystems 7500 FAST Real-Time PCR System . The commercially available primer probe sets  were adopted in PCR amplification. When genotyping was performed, appropriate negative controls were included in each run. Ten percent of participants were randomly selected as the masked duplicate sets to be genotyped twice by different persons, and the results were found to be concordant for all of them.2 tests were adopted to test the Hardy-Weinberg equilibrium (HWE) of the SNPs. was The odds ratios (OR) and their 95% confidence intervals (CI) to evaluate the associations between SNPs and oral cancer risks were calculated using unconditional logistic regression analyses. The association of miRNA polymorphisms and oral cancer risk stratified by sex were analyzed. The joint effects of miRNA polymorphisms and smoking on the risk of oral cancer were assessed by crossover analysis models. All of the tests were two-sided and statistical significance was defined as P<0.05. Multiple comparison tests (Bonferroni correction) were used and statistically significant P value threshold were defined as 0.017 after multiple comparison adjustment. All statistical analyses were performed using SPSS 20.0 software .The differences in distribution of demographic factors and tobacco smoking between cases and controls were examined using Pearson’s chi-square test and Student’s t-test. The goodness-of-fit χ2 = 0.314, P = 0.575). The smokers were more likely to develop OSCC . The distribution of demographic factors and tobacco smoking in cases and controls are shown in There are 340 cases and 340 controls included in the present study. The mean ages were 54.7±10.8 and 55.6±10.6 in case and control group and there was no difference in mean age between two groups . The chi-square tests showed the distribution of sex was equilibrium in cases and controls (XFurther analyses were performed stratified by sex, and more significant results were found in males but not in females . In maleIn addition, we examined the joint effects of miRNA polymorphisms and smoking on the risk of OSCC. As shown in In the present study, we evaluated the association between three functional polymorphisms in three common miRNAs and oral cancer risks in a Chinese population of 340 incident Asian OSCC cases and 340 healthy controls. To the best of our knowledge, this is the first study presenting the role of these three pre-miRNA polymorphisms together as well as their joint effect with tobacco smoking in the etiology of oral cancer in a northeastern Chinese population. Our findings suggest that the miR-499 rs3746444 polymorphism is associated with the risk of OSCC in Chinese, particularly among males.MiRNAs may act as regulators of gene expression and be involved in many cellular processes. Therefore, functional genetic polymorphisms of miRNAs may lead to individual variations in many cellular processes, leading to modification of the risk of cancers. The studies have shown that some miRNAs are up-regulated, while other microRNAs are downregulated in oral cancer –19. SuchRecently, several studies have reported the relationships of miRNA SNPs and the risk of head and neck cancers. One study indicated that the miR-196a2 rs11614913 polymorphism was associated with the risk of head and neck cancers . Liu et Because the frequency of the miRNA SNPs among the healthy controls is significantly different among different ethnicities, it is necessary to study the association of SNPs with cancer risk in diverse ethnic populations. So we underwent the present study in a Chinese population to reduce the risk of confounding due to population stratification and to provide the evidence of associations between miRNAs and oral cancer risks. Another major strength of the present study was the inclusion criteria for the controls. More and more evidence indicated that miRNAs were involved in some other diseases, which may be not clear. The control group in the current study included only healthy subjects, who were free of cancers or other chronic diseases, in order to exclude the effect of nonmalignant disease on the genotype distribution in the controls. In addition, because the age distribution of non-participating controls were similar to those of the participating ones, self-selection bias was unlikely. Moreover, the genotype distribution for each SNP was accordant with HWE, which further supports the randomness of our control subjects. Taken together, these strengths increased the reliability of our finding about an association between the miRNA polymorphisms and OSCC risks.In this case-control study, several limitations need to be addressed. Firstly, the cases and controls were chosen from hospitals and may not fully represent the general population which may result in selection bias in the study. So we should be cautious when we reached a conclusion. Secondly, because we selected only oral squamous cell cancer patients as our study population and excluded other head and neck cancer patients in order to control some confounding bias, the sample size could not be very large and statistical power of our study was limited, it is just as well that that the control group is as large as the case group; therefore it is possible to reach a more statistical power. Finally, the current study lacked the biological and functional research to determine whether the genetic variants identified in our studies modulate oral cancer risk through their influences on the functions or expressions of their host genes.In summary, this study firstly reported the association between single nucleotide polymorphisms in miRNAs and risks of oral squamous cell cancer in northeastern Chinese. Our study would have a certain positive significance in the understanding of oral cancer pathogenesis. Future larger studies with other ethnic populations and functional analysis are required to confirm current findings.The miR-146a rs2910164 and miR-499 rs3746444 polymorphisms might alter individual susceptibility to oral squamous cell cancer in Chinese.


Clostridium tetani neuro-toxin). (1) Emergency vaccination: while it has been very successful to reduce the impact of equine influenza epizooties and it is also recommended for tetanus in case of surgery and accident, the benefit of emergency vaccination against equine herpes virus 1/4 remains arguable; (2) Compatibility of equine vaccines from different brands: despite being a frequent concerns for equine veterinarians, little information is available about the compatibility of equine vaccines from different commercial origins. The consequence of mixing different equine vaccines targeting the same disease is believed to be limited but scientific evidences are sparse; and, (3) Laps vaccination and vaccine shortage: they could have serious consequences in terms of protection and their impact should be evaluated on a case by case basis, taking into account the risk of contact with the pathogen and the effect on herd immunity.To date, vaccination is one of the most efficient methods of prevention against equine infectious diseases. The vaccinology session, which was organised during the annual meeting of the French Equine Veterinarians Association (AVEF) at Reims (France) in 2016, aimed to approach three subjects of importance for the equine industry. Vaccination against three major equine diseases were used as examples: equine influenza (equine influenza virus), rhinopneumonitis (equine herpes virus 1/4), and tetanus ( Clostridium tetani (C. tetani) neuro-toxin).In many countries, the equine industry has a substantial economic weight  alone in 2013 . Ho. Ho26]. Mandatory EI vaccination in Hong Kong (preliminary result from a current study): around 30% of the horse population in Hong Kong is renewed every year (through importation). Equine influenza vaccination is mandatory for horses that are being exported to Hong Kong (in the weeks prior to exportation). Upon arrival, horses receive a fresh primary course of EI immunization with a unique EI vaccine. Due to numerous countries of origin and the diversity of EI vaccines used worldwide, the mix of EI vaccine in recently imported Hong Kong horses is inevitable. Preliminary results demonstrate that differences between pre- and post-importation EI vaccines has no measurable impact on EIV-specific SRH antibody response .In the absence of experimental data, sero-epidemiological studies provide some information. Results from two sero-epidemiological studies are summarised here:Several rhinopneumonitis vaccines are commercially available. EHV-1 and EHV-4 genetic and antigenic variability is believed to be relatively limited  . In thisTetanus vaccines contain an inactivated form of the tetanus toxin (toxoid). There are no antigenic differences between commercialised tetanus vaccines. As a consequence, tetanus vaccines are compatible, from an immunological point of view. However, vaccination schedules may differ between vaccines and brands, depending if the tetanus vaccine is formulated alone or in combination with another target . What is the immunological history (vaccination and/or infection) of the individual? Lapsed vaccination could be a concern if it happens during the primary course of immunisation or shorty after. It is likely to be less problematic in adult horses that have been frequently and repeatedly vaccinated. For example, the absence of a third immunisation (V3) against EI  may incrHow efficacious and immunogenic is the vaccine used? Consequences of lapsed immunisation is likely to be inversely proportional to the duration of immunity of the last vaccine administered.What is the level of herd immunity? If the number of animals with lapsed immunisation is limited, then the risk of infection will be counterbalanced by the level of herd immunity of the surrounding population .What is the risk of contact with the pathogen ? Is there time for emergency vaccination? For most vaccines, reaching protective levels of immunity will take a few days only after boost immunisation if the horse has been frequently immunised (prior to lapse).Lapsed vaccination happens when a vaccination schedule is not followed or maintained and in the case of a vaccine shortage. Lapsed vaccination should not be mistaken with immunity breakdown , which corresponds to a default of immunity  in an immunised animal (lapsed vaccination may result in immunity breakdown). For most vaccines and pathogen targets, the immunological consequences of lapsed vaccination depend on several key elements:In case of a vaccine shortage, veterinary practitioners have to face clients and horse owners discontent, which is justified by the concern of potential outbreaks . A vaccine shortage could induce a heavy financial loss .Both substitute products are monovalent EHV-1 vaccines. The number of doses available were limited , which inevitably led numerous French veterinary clinics to prepare new stocks in order to cover the requirement. However, the use of these stockpiles of monovalent rhinopneumonitis vaccines was a concern, especially in case of the renewed availability of the bivalent EHV-1/4 vaccine.Financial loss also concerned horse owners, due to more expensive substitute vaccines, the absence of commercial discount for these new products and the necessity to administer new primary immunisation courses for horses with lapsed vaccination history (in order to comply with race and event regulations).The shortage of bivalent EHV-1/4 vaccine did not result in a notable increase of EHV-4 outbreaks being reported to the French Epidemiological Surveillance of Equine Pathologies (RESPE) between 2015 and 2016 . HoweverFrom 2015 to early 2017, numerous European countries have been affected with a shortage of rhinopneumonitis vaccines. The two veterinary pharmaceutical companies providing these vaccines in Europe  experienced manufacturing issues with vaccine batch production and release, leading to a shortage of their rhinopneumonitis vaccines . Both companies obtained temporary authorisation to import and commercialise substitute EHV-1 vaccines . The welfare impact has been limited, but consequences for the French equine industry and veterinary practitioners were important:In the case of EI, emergency vaccination could significantly reduce the size and frequency of epizooties, as clearly demonstrated in South Africa (2003) and Australia (2007). However, the use of an EI vaccine with a DIVA marker is highly recommended. Several vaccines will reduce virus shedding but sterilising immunity is rarely observed. In this case, vaccinated horses may be infectious (duration of virus shedding is likely to be reduced when compared with unvaccinated horses). In this context, the implementation of other preventive measures  remains important. The usefulness of EHV emergency vaccination is more controversial, especially in naïve animals. In the case of tetanus immunization, emergency vaccination is essential to quickly neutralise the tetanus toxin.The compatibility of vaccines from different brands and manufacturers is a frequent question from veterinary practitioners and horse owners. Concerning EI vaccines, the question is mostly focused around the EIV strains contained in the vaccines. To date, available data to evaluate the immunological impact of mixed vaccination  remain limited. In any doubt, more frequent boost immunisations  may be advocated, especially after a change of vaccine. Serological analysis may prove useful to prevent any default of immunity . The immunological impact of lapsed immunisation or vaccine shortage is variable. The decrease in immunity is usually gradual with time passing since last immunisation (as opposed to the frequent public assumption of a sharp default of immunity). However, every breakdown in the vaccination schedule should be considered as potentially problematic, both at the individual and herd levels, and the consequences should be evaluated on a case-by-case basis.