Iron is generally not a free ion in the body, but is stored intracellularly by the protein ferritin and transported via association with transferrin. This iron sequestration is thought to occur because iron-mediated lipid peroxidation via Fenton chemistry can result from elevation in the free, or labile, iron pool. Cells in culture or in animals that experience iron overload can undergo cell death , 2, a pr2 .2. ISCs are critical components for at least 48 proteins including DNA polymerases, components of the electron transport chain, and several metabolic enzymes.Using RNAi-based loss-of-function screening in cell culture and tumor xenografts, we found that a significant number of shRNAs were differentially required when comparing survival in tumors versus cell culture, or at 21% oxygen versus 3% oxygen. These results led us to conclude that the difference in oxygen tension in tumor xenografts and cell culture is a major driver of differential essentiality. Two genes encoding critical steps of ISC biosynthesis, NFS1 and ABCB7, scored as top hits in both comparisons. NFS1 removes sulfur from cysteine for use in the synthesis of ISCs, cofactors which sensitive to damage by O2in vitro, growth at 3% O2, as orthotopic tumors in the mammary fat pad, or as subcutaneous xenografts was unaffected. Interestingly, breast cancer metastasis to the lung and primary lung tumor formation was significantly hindered by NFS1 knockdown. These observations led us to the conclusion that environmental oxygen concentration was driving the differential effects on tumor cell growth in the low oxygen environment of the mammary fat pad or subcutaneous space compared to the relatively higher oxygen environment of the lung. It is important to note that while established lung tumors are likely hypoxic, cells initially seeding lung metastases or pre-transformed lung airway epithelial cells may experience O2 concentrations near atmospheric levels. Consistent with this idea, analysis of human lung tumors revealed that NFS1 lies in a small genomic region undergoing positive selection in lung adenocarcinoma and NFS1 protein levels are elevated in well-differentiated tumors from this cancer type. These findings indicate that environmental oxygen levels experienced by incipient lung tumor cells can select for elevated NFS1 expression inase, an enzyme which can deplete serum cyst(e)ine levels thereby inducing oxidative stress, in a mouse xenograft model. Moreover, erastin treatment, which deprives cells of cysteine and therefore GSH produced from it, also results in a modest activation of the iron-starvation response. In accordance, NFS1 suppression strongly cooperates with inhibition of GSH biosynthesis to promote ferroptosis. These results are consistent with both ISC and GSH biosynthesis being downstream pathways inhibited by erastin and important for its induction of ferroptosis (Figure In the course of performing these experiments, we noted that adding pro-oxidant molecules to cells in which NFS1 had been suppressed resulted in cell death despite these treatments not resulting in a measurable ISC defect. These results suggested that a downstream consequence of NFS1 suppression rendered cells susceptible to death by exogenous oxidants. Interestingly, an ISC cofactor present in the iron responsive protein IRP1 functions as an iron-response element binding protein when the ISC is lost or damaged, activating the iron starvation response . AccordiBased on these findings, NFS1 suppression offers two therapeutic avenues for targeting cancer. Suppression of NFS1 inhibits lung tumor growth as a result of loss of cell essential functions from cofactor depletion, and at the same time activates the iron starvation response, priming cells to undergo ferroptosis in response to oxidant treatment.


Neurocritical care as a recognized and distinct subspecialty of critical care has grown remarkably since its inception in the 1980s. As of 2016, there were 61 fellowship training programs accredited by the United Council for Neurologic Subspecialties (UCNS) in the United States and more than 1,000 UCNS-certified neurointensivists from diverse medical backgrounds. In late 2015, the Program Accreditation, Physician Certification, and Fellowship Training (PACT) Committee of the Neurocritical Care Society (NCS) was convened to promote and support excellence in the training and certification of neurointensivists. One of the first tasks of the committee was to survey neurocritical care fellowship training program directors to ascertain the current state of fellowship training and attitudes regarding transition to Accreditation Council for Graduate Medical Education (ACGME) accreditation of training programs and American Board of Medical Specialties (ABMS) certification of physicians. First, the survey revealed significant heterogeneities in the manner of neurocritical care training and a lack of consistency in requirements for fellow procedural competency. Second, although a majority of the 33 respondents indicated that a move toward ACGME accreditation/ABMS certification would facilitate further growth and mainstreaming of training in neurocritical care, many programs do not currently meet administrative requirements and do not receive the level of institutional support that would be needed for such a transition. In summary, the results revealed that there is an opportunity for future harmonization of training standards and that a transition to ACGME accreditation/ABMS certification is preferred. While the results reflect the opinions of more than half of the survey respondents, they represent only a small sample of neurointensivists. Critical care as a dedicated medical subspecialty developed largely because of scientific and technological innovations which allowed the support of patients through catastrophic illness involving organ failure. Neurocritical care as a subspecialty of critical care began in the 1980s as physicians caring for critically ill neurologic patients recognized their unique challenges and formed dedicated intensive care units (ICUs) to optimize their care . Since tIn the United States, accreditation of training programs and certification of physicians are managed by non-governmental, non-profit, self-governed organizations. The Accreditation Council for Graduate Medical Education (ACGME) is the most influential of the training program accrediting bodies, while the member boards of the American Board of Medical Specialties (ABMS) are examples of individual physician certifying bodies. ACGME and ABMS boards require a critical mass of practitioners and specific milestones to confirm that a specialty is clearly defined, recognized, and self-sustaining. Before the 1980s, there was no certification offered in critical care medicine. In September 1980, the ABMS approved the multidisciplinary subspecialty of Critical Care Medicine, and beginning in the late 1980s, individual ABMS member boards provided certification in several critical care subspecialties with overlapping competencies but distinct scopes of practice .Although the foundation of neurocritical care as a valuable independent critical care subspecialty has been propagated by dedicated practitioners for over three decades, accredited training in this field is just completing its first decade. As a relatively new subspecialty, neurocritical care did not initially have the requisite membership and track record to be considered for accreditation and certification through the ACGME–ABMS system but, rather, was developed through the United Council for Neurologic Subspecialties (UCNS). The aim of the UCNS and similar organizations was to organize and structure subspecialties that were not yet prepared for inclusion by the ACGME–ABMS. The UCNS was launched in 2003 with the support of five parent professional organizations representing clinical neuroscience practitioners. The first certificates in neurocritical care were issued in 2007, and fellowship program accreditation followed in 2008.As of 2017, there were 1,240 UCNS-certified physician neurointensivists with diverse backgrounds including neurology, internal medicine, emergency medicine, and anesthesiology . ArguablGiven the growth and maturation of neurocritical care, accreditation through the ACGME–ABMS pathway is the subject of much discussion among neurointensivists. The Program Accreditation, Physician Certification, and Fellowship Training (PACT) Committee of the NCS was convened to support and promote excellence in training and certification of neurointensivists, and one of the first tasks of the committee was to review the current state of fellowship training. In 2016, a survey was developed by the PACT Committee and e-mailed to fellowship directors to ascertain the level of institutional support, training environment, and challenges faced at this stage of the field’s evolution. The PACT Committee specifically explored how the current UCNS pathway for accreditation of fellowship programs and certification of graduates was perceived and what program directors thought about the transition to the ACGME–ABMS pathway.Survey questions were compiled from ideas submitted by the members of the PACT Committee and addressed program accreditation, practitioner certification, institutional support, program director responsibilities, faculty and service structure characteristics, trainee characteristics, and training milestones. Respondents were provided opportunities to select categorical answers or numerical entries as well as to enter free-text comments or numbers. Once the committee members were satisfied with survey content, the survey was operationalized using Survey Monkey.Surveys were e-mailed to program directors of 54 of the 57 fellowship programs in existence at the time of the survey, and 33 (61%) of program directors queried completed the surveys. Survey questions and responses are shown in online Supplementary Material.Thirty-two of 33 (97%) respondents reported UCNS accreditation, while 12 of 31 (39%) reported concomitant CAST accreditation. Among the 12 institutions offering both UCNS-accredited and CAST-accredited neurocritical care fellowships, 2 (17%) had a common program director and 11 (92%) shared faculty.Of 32 respondents, 22% indicated that neurocritical care not being included in the ACGME–ABMS pathway may adversely affect candidate recruitment, and 35% felt that job opportunities available to graduating fellows may be adversely affected. However, 52% (15/29) felt that the ACGME–ABMS pathway would best facilitate integration of neurocritical care into the critical care mainstream in the future, and 68% (21/31) indicated that ACGME accreditation would be preferred as a vehicle for supporting future growth of neurocritical care as a field. Additionally, 69% (22/32) of respondents similarly indicated that ABMS certification was preferred, while 25% (8/32) preferred the UCNS and 6% (2/32) CAST, for future growth.Approximately half  of the responding program directors indicated that they receive institutional support, 30% (10/33) receiving protected time/effort and 18% (6/33) receiving a fixed stipend. The median designated effort reported was 8.5% (IQR 5–10).Slightly over half of respondents  reported having an administrative coordinator with at least a fractional Full Time Equivalent dedicated to the neurocritical care fellowship, but only 18% (6/33) received salary support from the institution for the administrative coordinator.Approximately three-quarters  of programs received institutional support for fellow salaries; one-third  utilized clinical revenue for fellow salaries. When institutional support for fellow salaries was provided, all fellows in the program were supported in 20/25 (80%) programs. Among these 20 programs which received salary support for all fellows in the program, the entire salary for each fellow was covered in 15 (75%), and only half of the salary was covered in the remaining 5 (25%). Among programs that received institutional support for fellow salaries, 59% reported that support provided to neurocritical care fellows was not different from that provided to fellows in ACGME-accredited programs at their institution. Three (9%) of 33 directors reported using clinical revenue to support fellows’ research projects.Fellowship directors were queried about current administrative responsibilities such as would be required of an ACGME-accredited fellowship Figure . While aThe median number of faculty associated with the training programs surveyed was five; nine programs had eight or more faculty members affiliated with the fellowship, while 18 programs had seven or fewer. Of the 223 faculty members affiliated with fellowships, 149 (67%) were UCNS-certified in neurocritical care. The most common subspecialty affiliation was neurology (68%), followed by anesthesiology (15%), pulmonary/internal medicine (6%), surgery (4%), neurosurgery (3%), and emergency medicine (3%).Sixty percent of respondents characterized their ICUs as “open,” with open units defined as those in which services other than neurocritical care admit patients and enter orders, in contrast to closed units where admission and order entry are under the sole purview of the neurocritical care service.The number of ICUs covered by the programs’ faculty and fellows ranged from one to seven with the majority covering one (53% of 28) or two (36% of 28). Forty two percent of programs included a step-down unit. In terms of the number of beds covered, the responses ranged from 8 to 54, with a median of 23. The question did not specify the type of beds, ICU or step-down, or whether overflow patients in other patient care areas could be included in the response. Most programs (78%) reported that attending physicians and fellows provided consultations outside of their parent ICU.Ninety-seven percent of programs included residents on the ICU team; 88% included acute care nurse practitioners; and 62% included physician assistants. Of those programs with advanced practice providers, 89% had advanced practice providers who were dedicated to the neurocritical care service. Thirty-two percent of the programs had night-time in-house attending coverage; the remainder had combinations of residents, fellows, and advanced practice providers in-house 24/7  participated in the San Francisco Match system,Sixty-two percent of programs (18/29) required applicants to complete an ACGME or Royal College of Physicians and Surgeons of Canada-accredited residency program. Twenty five of 28 (89%) programs supported J1 visas; 11/28 (39%) H-1B visas; and 9/28 (32%) O-1 visas. Seventy-two percent (21/29) offered 1-year training programs for neurosurgeons and candidates with critical care board eligibility/certification.Most of the programs  credentialed fellows as post-graduate trainees; the remainder credentialed fellows as faculty.Most program directors 19/28, 68%) indicated that procedural volumes should be mandated, but there was a wide variation as to procedural requirements and even whether specific procedures were required as part of training  most program directors favor the ACGME–ABMS pathway as a vehicle for future integration of neurocritical care into mainstream critical care; and (2) there is heterogeneity of institutional structures  and wide variation in procedural requirements among neurocritical care training programs.Almost all survey respondents directed UCNS-accredited programs, and while UCNS-certification was not thought to be detrimental to fellow recruitment and post-graduate careers, more than half of the respondents indicated that future acceptance and integration of the subspecialty could benefit from ACGME accreditation and ABMS certification. Overall, neurocritical care fellowship programs received less institutional support than comparable fellowships governed by ACGME. While most programs received salary support for fellows, only half received support for the director, and most did not receive support for the administrative coordinator. Per ACGME guidelines, such support would be mandated and would represent a shift from what is currently provided to training programs at many institutions . LikewisThe survey also revealed significant heterogeneity in fellowship training in neurocritical care related to differences in institutional structures as well as wide variation in fellow procedural requirements. While neurology is the major source of fellows, program faculty show a broader representation of backgrounds including a significant number from anesthesiology and other critical care subspecialties. Programs vary in whether they support foreign-trained or visa-sponsored trainees, something that would likely be standardized under ACGME. Although many programs accept candidates into 1-year pathways for neurosurgeons and those with prior critical care training, the percentage of graduates who have completed this track in recent years is unknown.Fellows train within both open and closed units, and work with residents, advanced practice providers, and faculty. Most coverage models have 24/7 in-house coverage, with fellows as over-night providers in 72%, most often without a night-time attending in-house. Approximately one-quarter of fellows are credentialed as attendings and can bill independently for E/M services and procedures. As previously discussed, with transition to the ACGME–ABMS pathway, revenue in programs where fellows are so credentialed could decline, as independent billing would no longer be permitted.While the heterogeneous background of faculty, trainees, and the institutional variations complicate the structure of neurocritical care training, they are not unique to neurocritical care. In fact, such heterogeneity is common among the currently recognized ACGME–ABMS disciplines .Variation was also the theme of fellow procedural competency requirements. Although some caution needs to be exercised in the analysis of the results due to different interpretations of “minimum volumes required,” most fellowship directors indicated that there should be specific requirements. Central venous catheter insertion and arterial catheter insertion appeared universally incorporated into fellowship training, but there was significant variability among programs in what was considered a minimum number required for competency. Procedures such as endotracheal intubation, thoracentesis, and intracranial procedures produced an even broader range of responses, from not being required to having various required minimums. These results highlight the current uncertainty around procedural requirements which could potentially lead to variable fellow competency on entry into independent practice. The results, however, also suggest an opportunity to derive consensus about procedural competency in neurocritical care which could lead to future standardization of requirements across training programs .ad hoc manner to 66 fellowship training programs currently, most which are formally accredited by the UCNS and, therefore, offer a pathway to UCNS physician certification. The current broad training requirements have allowed many institutions with diverse ICU structures and faculty to match and train fellows in neurocritical care, but given the current maturity level of the subspecialty, an opportunity may exist to standardize some of the training, such as procedural competency. The finding with the greatest potential implications for the subspecialty, however, is that more than half of the survey respondents believe that the ACGME–ABMS pathway is more desirable than the current UCNS pathway going forward. Caution needs to be exercised when interpreting this finding: while this survey represents more than half of neurocritical care fellowship directors, it contains only a small sample of all neurointensivists and may not be reflective of the attitudes of the field as a whole.In conclusion, the subspecialty of neurocritical care has transitioned from a few scattered programs accepting and training fellows in an All authors made substantial contributions to the design of the survey; analysis of the data; drafting, revision, and approval of the manuscript. All authors are accountable for the accuracy and integrity of the work. RD and VR contributed equally to manuscript preparation.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.


MEFV, responsible for familial Mediterranean fever has dramatically increased the understanding of the pathophysiology of AIDs, shedding light on the associated defective signaling pathways (Most cases of autoinflammatory diseases (AIDs) are characterized by attacks of systemic inflammation resulting in recurrent fever, acute arthritis, and increased acute phase proteins. Frequent symptoms are also skin rash, myalgia, and bone deformations. The appropriate diagnosis of an AID can be challenging due to the overlapping symptoms or the presence of comorbidities, as autoimmune diseases. The discovery of the first autoinflammatory gene, pathways . Since tNOD2 gene [responsible for Blau syndrome (BS)] was identified as a susceptibility gene. More recently, Jéru and colleagues describe the pathogenicity of a mutation affecting the same residue in TNFR1 in both Mendelian TRAPS syndrome and multifactorial inflammatory conditions   .Thus, not only a single gene could be involved in various phenotypes but also many different genes could be entailed in overlapping phenotypes.Consistent with this hypothesis, we propose that evolutionary gene duplication can have a causative role in AIDs development. Gene duplication is an important mechanism through which new genetic material is generated during molecular evolution. It can result from unequal crossing over, retroposition, or chromosomal/genome duplication . DuplicaA computational study by Wong and co-authors  observedWe have focused our attention on the nucleotide-binding domain, leucine-rich repeat (LRR) (NLR)-containing family, important components of the innate immune system. Only a few members of this family, including the cytosolic pattern recognition receptors NOD2, nod-like receptor protein 3 (NLRP3), and NLRP12, have been analyzed extensively. These NLRs are known to have a pivotal role in the innate immune response toward microbial invasion and are identfied as responsible for clinical distinct AIDs, which shares pathophysiological characteristics .NOD2 gene, encoding for the intracellular sensor of bacterial cell wall components (NOD2) that triggers the activation of NF-κB and MAPK pathways. NOD2 has a typical three-domain structure composed of two N-terminal effector regions consisting of a caspase recruitment domain, a centrally located domain NACHT that is critical for protein activation, and nine C-terminal LRRs that sense the pathogens. Mutations on LRRs are known to confer susceptibility to Crohn’s disease, a chronic inflammatory bowel disorder, whereas mutations in the central NACHT domain are associated with a granulomatous disease, named BS. BS is a rare autosomal dominant AID clinically characterized by granulomatous recurrent uveitis, dermatitis, and symmetric arthritis -1β to the activation of bacterial clearance processes.A group of autoinflammatory disorders called cryopyrin-associated periodic syndromes or CAPS results from mutations associated to the NALP3 inflammasome . CAPS enInterestingly, NLRP12 displays sequence and structure similarity to NLRP3 and plays a crucial role in immune system mechanisms against pathogenic agents . As CAPSin vitro to provoke spontaneous activation of the NLRP3 inflammasome  and NLRP3 and NLRP12 sequences (49.89% complete identity). Common clinical manifestations are also reported for the identical mutations, i.e., the absence of fever, the increased ocular involvement, and skin rash in the variations presented above. These findings suggest a close relationship between BS, CAPS, and NLRP12-associated disease, not only because both disorders affect the innate immune system but also because of common evolution.Investigation of the mechanisms that generate duplicate gene copies and the subsequent dynamics among gene duplicates represent a potential area for shedding light on evolutionary aspects related to interactions among all AIDs. From our point of view, the mechanism of gene duplication could help to explain the overlap of clinical symptoms and the great genetic heterogeneity in AIDs. We expect that the exponential increase in genomic data and rapid advances in molecular genetic technology will provide more evidence to support our hypothesis.This work was supported by Institutional funds (DOR1640251/16) from the University of Padova.GP contributed to the conception and to draft the work. Both PL and SP also contributed to revise it critically.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.


Despite the lack of evidence for symptomatic human infection with Maguari virus (MAGV), its close relation to Cache Valley virus (CVV), which does infect humans, remains a concern. We sequenced the complete genome of a MAGV-like isolate (OBS6657) obtained from a febrile patient in Pucallpa, Ucayali, Peru, in 1998. To facilitate its classification, we generated additional full-length sequences for the MAGV prototype strain, 3 additional MAGV-like isolates, and the closely related CVV (7 strains), Tlacotalpan (1 strain), Playas (3 strains), and Fort Sherman (1 strain) viruses. The OBS6657 isolate is similar to the MAGV prototype, whereas 2 of the other MAGV-like isolates are located on a distinct branch and most likely warrant classification as a separate virus species and 1 is, in fact, a misclassified CVV strain. Our findings provide clear evidence that MAGV can cause human disease. Aedes spp., Anopheles spp., Culex spp., Wyeomyia spp., and Psorophora spp.) in Ecuador, Brazil, Trinidad and Tobago, Colombia, Argentina, and French Guiana (Maguari virus (MAGV) was first isolated from a mixed pool of mosquitoes collected in Utinga forest in Brazil in 1957 and has since been isolated from a variety of mosquito species  reassortant, for which the name Itaya virus has been proposed , spanning 1957–1998 and originating from 3 of the 7 countries from which the virus has been isolated  . IsolateWe extracted viral RNA using the QIAamp viral RNA mini kit  according to the manufacturer’s instructions with reverse transcription PCR (RT-PCR) reactions performed using the Superscript III reverse transcription kit  and the iProof High-Fidelity PCR Kit . We obtained preliminary virus genome sequences of the MAGV BeAr7272 and AG83–1746 strains, as well as PLAV (strain 75V3066), TLAV (strain 61D240), and FSV (strain 86MSP18), using the 454 FLX pyrosequencing technology platform . We constructed libraries using previously described methods  Table 1;Phylogenetic and sequence divergence analyses were conducted on the open reading frames (ORFs) and their corresponding amino acid sequences. These analyses incorporated the sequences for MAGV, CVV, TLAV, PLAV, and FSV determined in this study and additional representative orthobunyavirus sequences available in GenBank  Table 2.Sequences for each ORF or corresponding amino acid sequence were aligned using the MUSCLE algorithm, and the evolutionary history for each tree construction was inferred using the neighbor-joining . Furthermore, an additional recombination analysis was performed on concatenated  genome sequences of all CVV, MAGV, PLAV, TLAV, and FSV isolates where full-length sequences were available. Initial alignment was performed by using the MUSCLE algorithm, as implemented in MEGA5 , the other comprises the AG83-1746 strain and the CbaAr426 strain . Both ofOur divergence analysis comparing the 2 groups formed by the isolates originally identified as MAGV showed 95.7% (N), 84.3%–85.2% (GPC), and 90.1%–90.7% (L) amino acid divergence. The extremely low level of N protein variability seems to be a characteristic of these and related virus groups, and the L protein divergence values are barely above the 10%-aa divergence sometimes suggested as a cutoff for speciation. However, the glycoprotein, which is the most variable of the viral protein products, differs substantially from all other related virus groups and also between these 2 groups. Furthermore, the consistent evidence for separate evolutionary histories inferred by the phylogenies for all 3 proteins indicates that these viruses most likely differ sufficiently to warrant reclassification of the AG83-1746 and CbaAr426 strains as a distinct virus species. In accordance with the prevailing naming conventions for bunyaviruses, which dictate naming based on the location of initial isolation, we use “Córdoba virus” (CODV) and “CODV lineage” herein to refer to viruses belonging to this group and to distinguish them from the canonical MAGV strains.In contrast to these various misidentified viruses, the OBS6657 isolate clearly grouped with the prototype MAGV strain BeAr7272 and showed 100% (N), 97.1% (GPC), and 99.0% (L) amino acid identity, respectively, across each of the three major gene products, despite being isolated >40 years later. Given that this virus was isolated from a patient exhibiting clear evidence of an acute febrile infection, our data provide strong evidence that MAGV infection can cause clinical disease in humans. In light of this evidence, and particularly when considered with serologic data that indicate frequent infection with MAGV-like viruses (2%–64% positive serum samples) . Taken together, the phylogenetic arrangement , malaise, muscle aches, and sore throat. The isolate cross-reacted very closely with CVV, MAGV, and PLAV virus in complement fixation but only with CVV and PLAV by plaque-reduction neutralization test  to the CVV group, and identification of FSV as a previously unrecognized CVV reassortant, is to substantially expand the known geographic range of CVV. On the basis of these data, the range of CVV must now be considered to include not only the United States and northern and central Mexico but probably all of Central America and the northern areas of South America. Furthermore, although both PLAV strains appear to belong to CVV lineage 1, TLAV belongs to CVV lineage 2, as does the CVV donor of the FSV M segment. This finding suggests that both lineages exist in Central America and that lineage 2 has been circulating in that region since at least the 1960s, when the first of these viruses were isolated. This supposition is also supported by the assignment of the MAGV CoAr3363 strain, which was isolated in 1964, to lineage 2. Taken together, these findings suggest that although these CVV lineage 2 strains have only recently emerged in the eastern United States, they might in fact have been circulating in South America for decades and have only recently been introduced into the United States from these regions.Our analysis of the ORF and amino acid data identified several distinct clades representing CVV (lineages 1 and 2) and 2 separate clades of what had been identified as MAGV strains . These cThe extent of this issue is further highlighted by our divergence analysis (In summary, we identified MAGV as the causative agent of a human febrile infection in Peru and showed that the virus associated with this infection is highly similar to the prototype MAGV isolate, suggesting that other viruses of this lineage also might have pathogenic potential in humans. In addition, our in-depth analysis of the closely related CVV group and other similar viruses showed that PLAV and TLAV do not exist as distinct virus species but are misclassified strains of other existing groups and that the human pathogenic FSV is a previously unrecognized reassortant between CVV, itself a known human pathogen, and a new lineage whose members had previously been identified as MAGV but are genetically distinct from the lineage occupied by the prototypical MAGV isolates. On the basis of its location of origin, we suggest the name “Córdoba virus” for this new lineage. Based on the reclassification of these virus isolates, it is clear that the endemic region for CVV is much larger than previously recognized and that CVV and MAGV most likely are responsible for unrecognized febrile infections throughout North Central, and South America. We hope that a better understanding of the genetic relationships between bunyavirus groups, particularly in relation to their relevance for human infection, coupled with an increased availability of reference genome sequence information, such as provided by this study, will help enable future surveillance and diagnostic efforts and increase awareness of the importance of orthobunyaviruses as human pathogens.Source and sequence information for Maguari virus strains sequenced in this study; GenBank accession numbers for reference orthobunyavirus sequences used in the phylogenetic analyses; comparison of phylogenetic analyses of nucleoprotein, glycoprotein and polymerase amino acid sequences; comparison of phylogenetic analyses of nucleoprotein, glycoprotein and polymerase open reading frame nucleotide sequences; divergence analyses for nucleoprotein, glycoprotein, and polymerase sequences.


Exocytosis is a fundamental cellular process whereby secreted molecules are packaged into vesicles that move along cytoskeletal filaments and fuse with the plasma membrane. To function optimally, cells are strongly dependent on precisely controlled delivery of exocytotic cargo. In mammalian cells, microtubules serve as major tracks for vesicle transport by motor proteins, and thus microtubule organization is important for targeted delivery of secretory carriers. Over the years, multiple microtubule-associated and cortical proteins have been discovered that facilitate the interaction between the microtubule plus ends and the cell cortex. In this review, we focus on mammalian protein complexes that have been shown to participate in both cortical microtubule capture and exocytosis, thereby regulating the spatial organization of secretion. These complexes include microtubule plus-end tracking proteins, scaffolding factors, actin-binding proteins, and components of vesicle docking machinery, which together allow efficient coordination of cargo transport and release. Exocytosis is a secretory trafficking process during which molecules are processed and transported to the cell surface, where they can be either released into the extracellular space or inserted into the plasma membrane. Secretory transport occurs in multiple steps: after budding from the Golgi, exocytotic vesicles travel along cytoskeletal filaments toward the cell periphery, come into contact with tethering factors that can restrain them, and subsequently dock and fuse with the plasma membrane with the aid of soluble NSF attachment protein receptors (SNAREs). Secretion can occur constitutively, to maintain cell homeostasis and provide components of extracellular matrix and cell adhesion structures (constitutive exocytosis). Alternatively, release of specific cargos in many types of differentiated cells can be tightly controlled in both space and time by a variety of signaling pathways (regulated exocytosis). Regulated exocytosis plays an important role in multiple processes, including synaptic neurotransmission, endocrine and paracrine signaling, or the release of hydrolytic enzymes by intestinal cells and leukocytes  to promote their remodeling and thus facilitate efficient cell movement , a heterogeneous class of proteins distinguished by their specific accumulation at the growing microtubule plus ends  domains and proteins with a short linear motif Ser-any amino acid-Ile-Pro (SxIP) embedded in unstructured positively charged regions  protein family22, was proposed to be involved in tethering microtubules to the cell cortex via IQGAP123, a cortical scaffold protein with interesting roles in exocytosis which we will discuss below -trisphosphate (PIP3)-binding protein LL5β36 10. Interestingly the ACF7-CAMSAP3 interaction was also associated with FA targeting and cell migration9.Finally, spectraplakins, a group of very large proteins, have been shown to be involved in cortical microtubule stabilization. Spectraplakins have the ability to directly link microtubules and actin filaments and are involved in a wide range of cellular processes. Despite the presence of only two mammalian genes, a variety of spectraplakin isoforms have been found. This results from the existence of alternative tissue-specific promotors and multiple splice variants  2/3 complex and formins. IQGAP can stimulate Arp2/3-dependent actin polymerization through direct as well as indirect interactions via the activation of neural Wiskott-Aldrich syndrome protein (N-WASp)77, has been shown to form a complex with IQGAP178. Remarkably, not only exocytosis but also endocytosis of insulin secretory membranes, a process essential to maintain a constant cell volume and to allow the reuse of exocytotic machinery, strongly depends on complex formation between Rab27A and IQGAP1. Depletion of IQGAP1 prevented glucose-induced redistribution of Rab27A from the cytosol to the plasma membrane78. These data suggest that IQGAP1 participates in both endocytosis and exocytosis upon glucose stimulation in β cells. Whether these functions relate in some way to the interactions of IQGAP1 with microtubule-binding proteins is currently unclear, especially as, strikingly, microtubules in β cells restrict, rather than promote, the availability of insulin granules for secretion79.Next to Rho GTPases, Rab GTPases also belong to the key regulators of membrane trafficking and exocytosis. Interestingly, Rab27A, a small GTPase that regulates exocytosis of insulin-containing vesicles in pancreatic β cells80 87 87 . As disc91, a transmembrane protein that is involved in axon guidance , the principal site of Ca2+-dependent exocytosis of neurotransmitters at neuronal synapses; these proteins thus have complex roles in neurotransmission across different animal species , and the ankyrin repeat protein KANK1. Liprin-α1 and -β1 were initially identified as interacting partners of the protein tyrosine phosphatase LAR87 102, an abundantly expressed Rab GTPase that strongly decorates the Golgi apparatus and cytoplasmic vesicles105. Although these vesicles were originally believed to be responsible for COPI-independent transport to the Golgi107, detailed imaging studies demonstrated that in fact they predominantly fuse with the plasma membrane and thus represent carriers of constitutive secretion101.ELKS is a ubiquitously expressed adaptor, which can be recruited to the plasma membrane by both LL5s and liprin-α1101. The underlying mechanism is not entirely clear. Similar to CAZ components, ELKS-containing complexes might promote the interaction between SNAREs located on the vesicles and the plasma membrane; however, it is currently unclear whether there is a direct connection between SNAREs and ELKS. Furthermore, in addition to the direct binding to ELKS, Rab6 also controls the recruitment to exocytotic vesicles of another Rab GTPase, Rab8A. Rab8A is a well-known player in exocytosis111. Interestingly, Rab8A interacts with ELKS-positive cortical sites through the binding partner of ELKS, MICAL3112, a multidomain oxidative enzyme which can promote disassembly of actin filaments and potentially remodel other protein complexes and also act as a scaffold114 , which can degrade the extracellular matrix around FAs, resulting in integrin detachment, loss of tension, and FA turnover116. These observations help to explain why liprin-α1, liprin-β1, LL5β, and ELKS promote invasive behavior and internalization of integrins in breast cancer cells119. Importantly, MT1-MMP delivery and integrin recycling also strongly depend on endosomal trafficking, which requires microtubules  are also formed at neuromuscular junctions (NMJs) undergoing remodeling during postnatal stages of development, and LL5β, which strongly localizes to regions of high density of acetylcholine receptors at the NMJ, has been implicated in this process126. At the NMJ, the complexes of LL5β and CLASPs were shown to capture microtubule plus ends and in this way create a route for the delivery of vesicles containing acetylcholine receptors to the postsynaptic membrane129. It is currently unknown whether ELKS participates in the regulation of the fusion of acetylcholine receptor-containing carriers with the plasma membrane, but this possibility seems quite likely, given the involvement of ELKS in secretion and the observation that ELKS is present at the NMJ130. Taken together, these data show that CMSCs or complexes related to them in composition regulate both microtubule organization and secretion in different types of undifferentiated as well as differentiated cells.LL5β and ELKS were also shown to concentrate at podosomes, actin-rich dynamic structures which can remodel the extracellular matrixin vivo they act in the same or different cell and tissue settings. Addressing this question will require systematic analysis of all major players using the same cellular models and also exploring their expression and interactions in tissues. Relevant in this respect is the analysis of tissue-specific isoforms of the investigated proteins. For example, whereas IQGAP1 was extensively studied, much less is known about its homologues IQGAP2, which is enriched in the liver and stomach, and IQGAP3, which is mainly found in brain and lung tissue131. Their domain composition is highly similar to that of IQGAP1 and given their specific expression in tissues with high exocytotic activity, they are interesting candidates for having profound but undiscovered roles in exocytosis.Microtubules play an essential role in exocytosis by serving as tracks for motor proteins that transport secretory carriers. The best studied mammalian cell models so far include migrating mesenchymal cells, in which a surprisingly diverse set of molecules is responsible for attaching and stabilizing microtubules to cortical sites close to the leading cell edge. An important unresolved question is whether the different complexes described so far, such as CMSCs and IQGAP-linked cortical assemblies, represent cooperating or redundant pathways or whether132. ELKS was also found to coincide with the docking and fusion sites of insulin in a pancreatic β-cell line; consistent with this observation, ELKS clusters show significant overlap with the clusters of the SNARE syntaxin 1, and ELKS depletion strongly affected insulin exocytosis133. However, as indicated above, the connections between CMSC components and SNAREs require further elucidation. Furthermore, microtubule plus ends do not appear to be directly connected to the sites of exocytosis in neurons or β cells. It is possible that microtubules are linked to secretory sites when relatively rapid transport of newly synthesized proteins from the Golgi apparatus is functionally important. Direct microtubule-based delivery might not be essential when an excess of secretory cargo is available or when extensive local recycling of secreted molecules takes place, as is the case in neurons. In some cell types, such as pancreatic β cells, microtubules may even be used to sequester rather than deliver secretory cargo79. Investigating the diversity of the mechanisms responsible for vesicle delivery and fusion represents an exciting subject for future research.Another interesting set of questions concerns the involvement of the discussed complexes in regulated secretion. There are strong data showing that many of the factors described above are important for regulated exocytosis. For example, ELKS and liprin-α are well-known players in neurotransmitter release in neurons and also are required for exocytotic release of inflammatory mediators by mast cells upon induction of allergic responses


A 39-year-old woman presented with a twice-recurrent, full-thickness left chest and abdominal wall metaplastic sarcomatoid carcinoma (MSC). Two additional radical resections of the tumor left a defect measuring 30 × 40 cm. An anterolateral thigh (ALT) free flap with anastomosis to the right gastroepiploic vessels was chosen for reconstruction.What are the local reconstructive options for chest and abdominal wall reconstruction?What are the free flap options?What considerations were made in the reconstruction of a patient with a large defect and vital organ exposure?Which vessels are used as a recipient in free tissue transfer in abdominal reconstruction?,2,6Advances in reconstructive surgery over the past 2 decades have permitted the oncologist to proceed with radical cancer resection and ensure negative margins without the fear of causing an irreparable defect. Local options in defect reconstruction include primary closure, skin grafting, tissue expansion, negative pressure-assisted closure, the components separation technique, prosthetic mesh, and pedicled flaps.1Free flaps are indicated when there is a massive loss of local tissues or when the defect size exceeds the reach or coverage of a pedicled flap.2a and b). In planning reconstruction, the extensive musculofascial damage and defect dimensions precluded our use of the rectus abdominis and the latissimus dorsi, respectively, so we chose to proceed with a free flap consisting of the contralateral ALT and a part of the vastus lateralis muscle. Incorporation of the vastus lateralis allowed for a larger skin paddle and its added bulk is beneficial for dead space obliteration.Fig 2c). The descending branch of the circumflex femoral artery was anastomosed to the right gastroepiploic artery through an opening on the posterior rectus sheath, over the liver, so as to avoid any undue tension or hernia formation (Our patient was a 39-year-old woman who presented with a twice-recurrent, full-thickness, left chest and upper abdominal wall MSC involving the costal margin and costochondral junctions of multiple lower ribs . The patFigs 2ormation  and 4.,8In abdominal reconstruction, recipient vessels typically utilized are the extraperitoneal vessels, including the deep epigastric vessels, the internal mammary vessels, and the saphenous vein loop graft; however, intraperitoneal vessels, such as the gastroepiploic vessels, may also be used.7Successful reconstruction is contingent on fully addressing the anatomical, functional, and cosmetic deformities of a defect. Our case demonstrated reconstruction of a complex wound in a single stage using the gastroepiploic artery for anastomosis.


Laboratory models have been crucial to increase knowledge about oncogenic mechanisms but due to the limitations of rodents and cell culture approaches, they are viewed as artificial models, whereas the spontaneous tumors in pets are empirically considered to be a more powerful and reliable tool. With the publication and full annotation of cat's genome , researcNowadays, the incidence of feline mammary carcinoma (FMC) is increasing, probably because of the humanized lifestyle of the cat, unregulated environmental factors and better prevention/efficient treatment of infectious diseases. Despite this scenario, few studies have evaluated the protein expression in FMC, in order to uncover new prognostic factors and discover possible therapeutic targets. In two recent studies, we identified in a pool of FMC, the six molecular subtypes described in breast cancer patients, using the broadly accepted IHC-based classification established by St. Gallen International Expert Consensus Panel ,3. The rConcerning the role of the feline homologue of human epidermal growth factor receptor-2 (HER2) in mammary carcinoma, some reports have brought fragmentary data revealing that the molecular mechanisms underlying HER2 tumorigenesis may not be fully conserved between cat and man. The overexpression of this transmembrane glycoprotein that belongs to the epidermal growth factor receptor family, is correlated with a rapid tumor growth and progression in breast patients and because of this, is routinely measured by immunohistochemistry and by in situ hybridization, since in the majority of the cases (80%), HER2 overexpression is associated with gene amplification. Published data reveal that 20-30% of human breast cancer patients display HER2 over-expression, prompting a therapy based on humanized monoclonal antibodies and tyrosine kinase inhibitors which improves survival rates. In cats, although the majority of HER2-positive mammary carcinomas are classified as either tubulo-papillary or solid adenocarcinomas, as human HER2-positive breast tumors and the incidence of HER2 overexpression in mammary carcinomas 20-40%) seems to be similar to the one that occurs in breast patients 0% seems ,6, beingRecently, five sequence variations in the HER2 gene region that encodes for the extracellular domain, comprising exons 17 to 20, were described with 2 non-redundant sequence variations being associated with malignant/metastatic-status specificity . InteresIn summary, the published data clearly show that more research on HER2-positive FMC is needed in order to fully prove that this pathology is a suitable cancer model. From the veterinary point of view, the high prevalence of HER2 overexpression in FMC can be a great opportunity to study specific target therapies directed against the HER2 receptor and to improve diagnostic tools based on tissue and serum HER2 status.


The prefrontal cortex (PFC) is indispensable for several higher-order cognitive and executive capacities of primates, including representation of salient stimuli in working memory (WM), maintenance of cognitive task set, inhibition of inappropriate responses and rule-guided flexible behavior. PFC networks are subject to robust neuromodulation from ascending catecholaminergic systems. Disruption of these systems in PFC has been implicated in cognitive deficits associated with several neuropsychiatric disorders. Over the past four decades, a considerable body of work has examined the influence of dopamine on macaque PFC activity representing spatial WM. There has also been burgeoning interest in neuromodulation of PFC circuits involved in other cognitive functions of PFC, including representation of rules to guide flexible behavior. Here, we review recent neuropharmacological investigations conducted in our laboratory and others of the role of PFC dopamine receptors in regulating rule-guided behavior in non-human primates. Employing iontophoresis, we examined the effects of local manipulation of dopaminergic subtypes on neuronal activity during performance of rule-guided pro- and antisaccades, an experimental paradigm sensitive to PFC integrity, wherein deficits in performance are reliably observed in many neuropsychiatric disorders. We found dissociable effects of dopamine receptors on neuronal activity for rule representation and oculomotor responses and discuss these findings in the context of prior studies that have examined the role of dopamine in spatial delayed response tasks, attention, target selection, abstract rules, visuomotor learning and reward. The findings we describe here highlight the common features, as well as heterogeneity and context dependence of dopaminergic neuromodulation in regulating the efficacy of cognitive functions of PFC in health and disease. The prefrontal cortex (PFC) is a critical node in brain networks involved in complex cognitive functions and has undergone a great evolutionary expansion in primates Wise, . PFC dysThe PFC is subject to substantial neuromodulation from all major ascending modulatory systems  family comprising of D1 and D5 receptors and the Gi-coupled D2 receptor (D2R) family comprised of D2, D3 and D4 receptors  in neighboring periprincipal PFC, affecting neurons with identical delayed response activity, increased contralateral memory-guided saccade errors and reaction times (Sawaguchi and Goldman-Rakic, Another, related point of interest concerns the physiological actions of dopamine. Data gathered in macaques from a series of iontophoretic studies (Wang et al., A trivial explanation would be that these differences in behavioral consequences of D1R blockade reflect an areal difference in dopamine’s modulation of the “spotlight of attention”. Alternatively, there may be differences in dopaminergic modulation of  “immediate” vs. “delayed” selection of action targets: enhancement of response field stimulus selection could be positively modulated by D1R blockade when the stimulus is visible, whereas the introduction of a delay would cause the opposite effect, perhaps due to increases in variability of cortical representations in the intervening delay period leading to deterioration in attentional representation of the stimulus location. These divergent pharmacological effects appear to highlight the heterogeneity of D1R actions on different cortical areas. Future experiments employing similar microinjections in a task similar to Lebedev et al.  may shedDopamine has a well-established role in reward signaling in the brain (Schultz et al., gain of spatial tuning in the memory period, a phenomenon reminiscent of attentional modulation. Interestingly, Leon and Shadlen (Ventrolateral PFC neurons are modulated by reward, and spatial selectivity of neurons therein is modulated by the reward contingency (Kennerley and Wallis,  Shadlen  found th Shadlen , and the Shadlen . These f Shadlen .Two studies examined the role of PFC D1Rs and D2Rs in associative learning (Puig and Miller, Puig and Miller , 2014 alThus, injections of antagonists of both dopamine receptor families disrupted novel visuomotor learning involving associating objects with saccades in particular directions, but did not affect familiar and well-learned associations. This is interesting when considering effects of injection of these antagonists in periprincipal PFC on oculomotor delayed response performance, as discussed in the previous section (Sawaguchi and Goldman-Rakic, Another critical role that has been associated with PFC is in the maintenance of cognitive sets: learning, maintenance and application of sets or rules used to guide flexible responses to environmental stimuli (Wallis and Miller, PFC neurons maintain information about the cognitive set and the rule to be employed in a behavioral trial (Wallis et al., Recently, our group examined the influence of dopamine receptor subtypes on rule representation in PFC (Vijayraghavan et al., Using iontophoresis, we examined the effects of D1R and D2R stimulation on the activity of dorsolateral PFC neurons engaged in this task. We tested two agonists, SKF81297 and quinpirole, and examined the overall effects and dose-dependence on PFC excitability and task representation. D1R stimulation had an overall suppressive effect on the population of PFC neurons tested, while D2R stimulation had an overall excitatory effect Figure , consistOur results with dopamine receptor modulation of rule selectvity were different from those found in the study by Ott et al. . One expIn our study (Vijayraghavan et al., A previous study found that D2Rs selectively modulated the activity of neurons that were only activated during the saccade response in the spatial delayed response task (Wang et al., What then are the origins and functional role of perisaccadic activity in dorsolateral PFC, and thereby, what is the role of D2Rs in the neuromodulation of this activity?One possibility is that, D2Rs modulate the presaccadic component of perisaccadic activity that is involved in saccade target selection. PFC neurons exhibit greater representation of the contralateral visual space in their stimulus-driven activity (Funahashi et al., In this review, we have described the gamut of evidence about the role that dopaminergic neuromodulatory systems play in regulating the cognitive functions of PFC in primates. While much work has been done over the past four decades on involvement of dopamine signaling in shaping PFC cognitive circuitry during delayed response task performance, new studies performed over the past decade have begun to explore the role that dopamine plays in other cognitive and executive functions in which the PFC is intimately involved, including spatial attention, maintenance and flexible switching of cognitive sets, reward and feedback representation. While consensus is emerging about the direct physiological effects of dopamine receptor subtypes in the PFC of awake behaving primates, the complexity of dopamine’s actions on PFC task activations point to heterogeneity in the functions of cortical dopamine. Further work examining potential links between what is known about dopamine’s role in reward and reinforcement learning and its modulation of “states” of activity in well-learned tasks are needed. Such studies will be of considerable significance, given the interest in dopaminergic pharmacology in the development of treatments for enhancing cognitive outcomes in patients with neuropsychiatric diseases.SV and SE designed the experiments. SV and AJM conducted experiments and analyzed data. SV, AJM and SE wrote 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.


Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophic factor family of signaling molecules. Since its discovery over three decades ago, BDNF has been identified as an important regulator of neuronal development, synaptic transmission, and cellular and synaptic plasticity and has been shown to function in the formation and maintenance of certain forms of memory. Neural plasticity that underlies learning and memory in the hippocampus shares distinct characteristics with spinal cord nociceptive plasticity. Research examining the role BDNF plays in spinal nociception and pain overwhelmingly suggests that BDNF promotes pronociceptive effects. BDNF induces synaptic facilitation and engages central sensitization-like mechanisms. Also, peripheral injury-induced neuropathic pain is often accompanied with increased spinal expression of BDNF. Research has extended to examine how spinal cord injury (SCI) influences BDNF plasticity and the effects BDNF has on sensory and motor functions after SCI. Functional recovery and adaptive plasticity after SCI are typically associated with upregulation of BDNF. Although neuropathic pain is a common consequence of SCI, the relation between BDNF and pain after SCI remains elusive. This article reviews recent literature and discusses the diverse actions of BDNF. We also highlight similarities and differences in BDNF-induced nociceptive plasticity in naïve and SCI conditions. BDNF was purified as the second member of the neurotrophin family in 1982 by Barde et al. i is high, NKCC1 is highly expressed, thereby mediating the excitatory action of GABA seen during early development. However, in adult neurons, the inhibitory actions of GABA are due to a predominant KCC2 expression. GABA receptor functions and chloride homeostasis are influenced by manipulations that modify the expression of NKCC1 and KCC2. In hippocampal cells, BDNF decreased both the expression and phosphorylation of KCC2 [The multifaceted BDNF-GABA relationship after peripheral and spinal cord injury is directly linked to the ability of both BDNF and SCI to independently regulate the expression and function of chloride transporters. K of GABA , 172, un (NKCC1) , 174  in intact spinal cord, after SCI, BDNF could produce protective or antinociceptive effects by increasing KCC2 and GABAergic inhibition. Clearly, there are no solid explanations for these differential effects of BDNF or the mechanisms that trigger a tangible switch in BDNF functions after SCI. Both small diameter primary afferents and activated microglia are likely sources of increased levels of BDNF within the spinal cord. If high levels of BDNF are retained into the chronic stages of injury presumably after the primary injury has been healed, then probable mechanism of maintained pain hypersensitivity would be BDNF-induced reduction in KCC2 mediated GABAergic inhibition. However, although the effect BDNF has on KCC2 and GABA circuits in the chronic stage of injury is clearly speculative at this time, it seems that SCI-induced reduction in KCC2 expression is considered a possible mechanism that underlies acute or early onset pain after SCI.Nonetheless, different observations are noted after SCI. Boulenguez et al.  recently per se. It is not determined whether these changes in ERK expression are determined by BDNF-TrkB signaling or whether BDNF-induced activation of these neural substrates plays an important role in producing neuropathic pain after SCI particularly as ERK can be engaged by other signaling pathways such as TNFα. Moreover, as we and others have shown, both TNFα and its receptors (TNFR1 and TNFR2) are upregulated after SCI [As we discussed earlier, ERK-dependent signaling mechanisms are involved in the development of pain. Interestingly, in our studies, we observed a decrease in ERK1/ERK2 and activated ERK1 (pERK1) expression at the lesioned epicenter during the first 7 days after SCI  althoughfter SCI  and may Importantly, it should be noted that although SCI appears to mimic some of BDNF's detrimental actions, these effects may not be mediated by BDNF itself. The phenomenon termed “metaplasticity” by Abraham and Bear  is a chaStudies of spinal plasticity have revealed that the processes involved are indeed complex. An even greater level of complexity arises after SCI, partly due to the enormous amount of cellular and morphological changes that ensue. BDNF has been classified as a neuromodulator and has been shown to be intricately involved in spinal plasticity. Clearly, there is no uniformity with BDNF actions as it has been shown that it can mediate both adaptive plasticity and maladaptive plasticity in spinal networks. For many years, our studies focused on unraveling these differential effects BDNF has on spinal networks in uninjured and injured spinal cord. Deductions made from several studies including some from our laboratories overwhelming show that as it relates to nociceptive processes, whereas BDNF exerts pronociceptive actions in the absence of injury, no such effect is observed after injury, at least acutely. Moreover, we found that functional deficits and mechanical sensitivity after SCI are associated with reduced BDNF levels, not increases. In contrast, increases in BDNF after SCI promote adaptive plasticity and functional recovery. The multifaceted effects of BDNF after SCI may be linked to SCI-induced changes in (i) expression of TrkB receptor and downstream kinases, (ii) GABA transmission due to altered expression of chloride transporters, and (iii) the activity of astroglial cells. The idea that BDNF plasticity is potentially susceptible to SCI-induced modifications indicates that metaplastic changes are essential to spinal function, particularly after injury to the CNS. In conclusion, we show that while significant progress has been made towards a clearer understanding of BDNF functions, after 30 years, additional research is warranted to elucidate the spinal effects of BDNF-TrkB signaling.


To evaluate the vascular changes of idiopathic macular telangiectasia type 2 (MacTel 2) patients with optical coherence tomography angiography (OCTA) and correlate these changes with the findings of spectral domain optical coherence tomography (SD-OCT).Simultaneous SD-OCT and OCTA images of 10 eyes of 6 patients who were diagnosed as MacTel 2 in Ankara University Faculty of Medicine, Department of Ophthalmology were obtained and graded according to the OCTA grading system for MacTel 2.Ten eyes of 6 patients were included. Four (66%) patients were female and 2 (34%) were male. The disease was grade 0 in 2 eyes, grade 1 in 2 eyes, grade 2 in 3 eyes, grade 3 in 1 eye, grade 4 in 1 eye, and grade 5 in 1 eye. The most common findings in grade 1, 2, and 3 non-proliferative disease were thinning of the outer retinal layers, presence of intraretinal hyporeflective layers and inner limiting membrane draping. In cases with subretinal choroidal neovascularisation (CNV) in OCTA, CNV or CNV scar was present in the B-scan SD-OCT images. In a case in which OCT was within normal limits, vascular changes consistent with grade 1 disease were observed in OCTA. On the contrary, 2 patients with significant foveal atrophy and macular hole in B-scan showed changes of early disease in OCTA. In some of the eyes, OCTA revealed an intact superficial vascular layer despite visible changes in the deep layer and the presence of neovascularisation.OCTA yields findings which are important for understanding the pathogenesis of the disease and providing better follow-up. Contrary to fundus fluorescein angiography, changes in the deep arterial plexus in the early disease and CNV can be clearly observed with OCTA. To achieve the best results in clinical practice, en face flow maps should be evaluated together with B-scan SD-OCT images. It is the most common type of IJRT, and includes cases that are usually bilateral, occult, and nonexudative. The disease typically presents in the fifth and sixth decades and affects men and women equally, although various studies have reported contradictory findings on the latter point.5Idiopathic macular telangiectasia type 2  is an acquired vascular disease of the macula primarily involving neural and glial cell degeneration and loss. This condition, named by Yannuzzi et al.,6 Optical coherence tomography (OCT) allows better understanding of retinal pathologies. Characteristic findings of the disease include macular thinning, hyporeflective cavitations in the inner and outer retinal layers, development of full-thickness macular hole in the absence of vitreoretinal traction, and atrophic changes in the outer retinal layer.8Clinically, it is characterized by loss of transparency in the foveal region, intraretinal crystalline deposit accumulation, hyperplastic retinal pigment epithelium migration, macular pigment loss, and progressive abnormalities in the juxtafoveolar retinal vessels. These include right-angle veins, subretinal and outer retinal neovascularization, and vascular invasion of the foveal avascular zone.3From its initial description to the present day, there has been no consensus regarding the mechanism of perifoveal telangiectasia. However, it is currently believed that the degeneration of Müller cells, which serve protective and supportive functions, is the primary pathology, and the damage to the retinal layers, atrophy, and vascular changes develop secondarily.9Optical coherence tomography angiography (OCTA) is an imaging method that visualizes the movements of blood cells using motion contrast, thus providing information about blood flow. Thusly, imaging of the retinal vascular layers is possible without the need for intravenous contrast injection. As opposed to fundus fluorescein angiography (FFA), in which layers other than the superficial vascular plexus appear only as background hyperfluorescence, OCTA enables separate imaging of the superficial and deep vascular layers and the choroidal vessels.In this study, we aimed to use OCTA to evaluate vascular anomalies in patients with MacTel 2 and elucidate the relationship between these and retinal anomalies detected on OCT.10  and OCTA images were recorded simultaneously from 6 patients diagnosed with MacTel 2 in the Ophthalmology Department of the Ankara University Faculty of Medicine. Patients were considered to have MacTel 2 in the presence of findings such as reduced autofluorescence in the central macula on fundus autofluorescence; loss of retinal transparency, intraretinal pigment, presence of right-angle venules, and telangiectatic vessel appearance on fundus examination; and OCT showing interior limiting membrane drape, retinal pigment epithelium migration creating intraretinal hyperreflectivity, atrophy of the outer retinal layers, and presence of intraretinal hyporeflective cavitation. OCTA en face flow maps were created using RTVue XR Avanti version 2015.1.1.98 and Split Spectrum Amplitude Decorrelation Angiography images were obtained. Images that could not be evaluated due to low resolution were excluded. Each eye was graded based on the MacTel OCTA grading system according to Chen et al.10 .Ten eyes of 6 patients were included in the study. Four (66%) of the patients were female, 2 (34%) were male. Their mean age was 67.6 years. The OCT and OCTA findings of the patients are shown in 9 demonstrated that FFA primarily shows the superficial vascular network of the retina and is inadequate for visualizing the deep capillary layer and the choroid. Consistent with other OCTA studies, in our series we observed that the superficial vascular network of some MacTel 2 patients was remarkably well preserved, even in advanced disease, while pronounced changes were evident in the deep capillary network. In addition, abnormalities may occur in the deep vascular network even in early disease stages where OCT is within normal limits, and FFA alone is not sufficient to evaluate these patients. It has also been shown that FFA findings which may be evaluated as perifoveal vascular leakage and lead to unnecessary injections in clinical practice are due to telangiectatic vessels in the deep vascular layer.11 OCTA clearly demonstrates that leakage simulating choroidal neovascularization on FFA is not a result of the formation of new vessels. Other advantages over FFA are that it is easy to perform, can be repeated frequently, and is non-invasive.OCTA provides valuable information regarding disease severity and neovascularization activity in patients with parafoveal telangiectasia. In a study using OCTA, Spaide et al.13 Müller cells are known to play important roles in the maintenance of foveal structural integrity, neuronal support, and continuity of the blood-retina barrier.15 It is believed that macular hole and intraretinal cavitation arise due to neural atrophy and disruption of the foveal structure resulting from the destruction of Müller cells.13 These findings appear independent of vascular changes and show the clinical importance of en face circulation maps in combination with B-scan OCT.In the current series, OCTA evaluation of patients with advanced atrophic changes in the foveal region and macular hole revealed minimal changes consistent with grade 0 or 1 disease in the foveal vascular network. These findings support the theory that retinal structural changes seen in MacTel 2 are not secondary to vascular anomalies, but are a primary condition.16 studied patients with marked MacTel 2 in one eye and apparently healthy fellow eyes and detected foveal pit asymmetry and temporal foveal thinning which were associated with very early disease.On the other hand, a patient with grade 2 disease in the left eye exhibited no pronounced abnormality in the right eye on OCT, while OCTA revealed reduced vascular density and telangiectatic vessels in the deep vascular plexus. On careful examination of SD-OCT images of this eye, it was noted that the foveal pit was asymmetric. Similarly, Charbel Issa et al.18 In their OCTA studies of MacTel 2, they emphasized the antiangiogenic properties of Müller cells, stating that hypoxia due to deep vascular network dysfunction and suppression of antiangiogenic factors due to Müller cell degeneration may lead to intraretinal neovascularization. Several previous studies have indicated that neovascularization in MacTel 2 originates more from retinal vascular structures rather than the choroid.20 In their study using OCTA, Zhang et al.21 showed that these subretinal neovascular complexes may not be only retinal, but may also be connected to the choroidal vasculature.Another finding we observed in our cases is that the formation of new vessels was not limited to the choroid, but could also occur between the outer retinal layer and the choroidal vascular network, which is usually avascular. This finding is also similar to those observed by Spaide et al.22 Spaide et al.18 also emphasized the importance of Müller cells in intraretinal vascular circulation and retinal support. Thus, it seems that Müller cell loss is responsible not only for intraretinal neural cell death and morphologic changes, but also vascular telangiectasia, leakage, and neovascularization.The fact that the severity of intraretinal and vascular changes are mutually independent supports the view that the primary pathology of MacTel 2 is not vascular. Moreover, it has been shown that Müller cell loss may be another factor that leads to telangiectasia.The results of this pilot study demonstrate that OCTA, which has recently been introduced into clinical use, offers insight into the pathogenesis of MacTel 2 and is useful as an auxiliary method to OCT in patient follow-up, but does not directly correlate with OCT findings in terms of the extent of anatomic abnormalities.


The aetiology of obesity has been attributed to several factors ; however none of these fully explain the increase in the prevalence of obesity worldwide. Gut microbiota located at the interface of host and environment in the gut are a new area of research being explored to explain the excess accumulation of energy in obese individuals and may be a potential target for therapeutic manipulation to reduce host energy storage. Several mechanisms have been suggested to explain the role of gut microbiota in the aetiology of obesity such as short chain fatty acid production, stimulation of hormones, chronic low-grade inflammation, lipoprotein and bile acid metabolism, and increased endocannabinoid receptor system tone. However, evidence from animal and human studies clearly indicates controversies in determining the cause or effect relationship between the gut microbiota and obesity. Metagenomics based studies indicate that functionality rather than the composition of gut microbiota may be important. Further mechanistic studies controlling for environmental and epigenetic factors are therefore required to help unravel obesity pathogenesis. Initial Evidence of the Role of Gut Microbiota in Obesity. The worldwide increase in obesity has prompted researchers to investigate its aetiology which is multifactorial, involving environmental, dietary, lifestyle, genetic, and pathological factors. Although the gut microbiota were already established as a metabolic organ that could ferment nondigestible dietary components (particularly nondigested carbohydrates) to generate short chain fatty acids (SCFA), their role as a significant environmental factor affecting host adiposity through an integrated host signalling pathway was explored in 2004 by Bäckhed and colleagues [ de novo lipogenesis and triglyceride storage through carbohydrate response element binding protein (ChREBP) and sterol response element binding protein 1 (SREBP1) and by suppressing fasting induced adipocyte factor (fiaf) which is an inhibitor of adipocyte lipoprotein lipase [lleagues . This br ob genotype in genetically obese leptin deficient C57BL/6J ob/ob mice and lean mice (ob/+ and +/+ wild-type siblings) fed similar polysaccharide rich diets, the ob/ob mice had reduced relative abundance of Bacteroidetes (by 50%) and a proportional increase in Firmicutes regardless of kinship [Subsequent studies conducted by the same group suggested that although gut microbiota communities were shared between mothers and offspring regardless of kinship . A highe ob/ob mice clustered together , richer in enzymes for degradation of polysaccharides, higher faecal acetate and butyrate, and less stool energy loss than in lean mice. Transplantation of gut microbiota from ob/ob mice or lean mice to germ-free mice resulted in obese (high Firmicutes) or lean (high Bacteroidetes) gut microbiome in the recipients. Obese microbiome recipients had higher percentage body fat despite similar food intake.Turnbaugh et al. (2006) used whole genome shotgun metagenomic and microbiota transplantation studies to investigate the mechanisms . They obIn a human study , obese a fiaf increased in HF and WD fed GF mice compared to CONV mice but not in the systemic circulation [Further evidence suggested the presence of the gut microbiota was necessary for development of obesity as germ-free mice were resistant to obesity even when they consumed more calories from normal chow or a high fat Western-type diet compared with CONV mice . Howeverculation .Several possible mechanisms were proposed to explain the impact of structural and functional differences in gut microbiota in lean and obese individuals that may contribute to host adiposity and whether an obese phenotype is transmissible by transplantation of gut microbiota. However, most of these studies were conducted in experimental animals which exhibited different anatomical, physiological, and bacterial colonisation patterns from humans. Several human and animal based studies have now revealed controversial evidence attributing differences in gut microbiota to the differences in diet –17 whileThe gut microbiota can be regarded as a “microbial organ” contributing to a variety of host metabolic processes from digestion to modulation of gene expression. The differences in gut microbiota between lean and obese animals or human subjects suggest a link between gut microbiota and energy homeostasis although there is still some debate as to whether these differences are causally related to an obese or lean phenotype. Various mechanisms have been suggested to link gut microbiota with obesity-genesis and other metabolic disorders . HoweverDietary polysaccharides and proteins that escape digestion in the small intestine are fermented in the colon by the gut microbiota into SCFA mainly acetate propionate and butyrate. The amount of energy harvested is hypothesised to be influenced by the composition of the gut microbiota . It has To a greater extent, there is a general agreement from many studies that the obese phenotype is associated with excess SCFA in caecal and faecal samples in animal and human studies compared with the nonobese . HoweverThe obese phenotype in animals is associated with higher total caecal SCFA, acetate, and butyrate and higher expression of bacterial genes responsible for polysaccharide metabolism . IncreasFiaf/ANGPTL4) is a target gene for peroxisome receptor activated proteins (PPARs) and is produced by large intestinal epithelial cells and the liver. Fiaf/ANGPTL 4 inhibits lipoprotein lipase (LPL) which causes accumulation of fat in peripheral tissues. Inhibition of fiaf by the gut microbiota with a resultant increase in LPL may be one mechanism for gut bacterial induced host adiposity [ fiaf genes (fiaf −/−). Lack of the fiaf gene causes disinhibition of LPL which leads to deposition of up to 60% higher epididymal fat compared to germ-free wild-type littermates expressing fiaf genes (fiaf +/+). fiaf/ANGPTL4 is therefore involved in the regulation of fat storage mediated by the gut microbiota. Controlled manipulation of the gut microbiota may alter the expression of this hormone [ Lactobacillus paracasei F19 for 10 weeks. Compared to the nonsupplemented group, plasma fiaf/ANGPTL4 was upregulated in the Lactobacillus paracasei F19 supplemented group with significantly elevated plasma VLDL but no change in other lipoproteins. In another study, Lactobacillus paracasei F19 and Bifidobacterium lactis BB12 were found to upregulate ANGPTL4 in the colon carcinoma HCT116 cell line in a dose and time dependent manner while Bacteroides thetaiotaomicron had no effect [ fiaf/ANGPTL4 is dependent on the gut microbiota and future interventional studies on weight management can be based on modification of ANGPTL4 by manipulating the gut microbiota.Fasting induced adipocyte factor or angiopoitein-like protein 4 , an enzyme in liver and muscle cells that acts as a fuel gauge monitoring cellular energy status. Inhibition of AMPk results in reduced muscle and liver fatty acid oxidation ultimately leading to excess fatty acids storage in these tissues .Phosphorylated AMPk inhibits the formation of malonyl CoA via acetyl CoA carboxylase. Inhibition of malonyl CoA causes disinhibition of carnitine palmitoyltransferase-1 (Cpt-1) which in turn catalyses the rate limiting step in the entry of long chain fatty acyl-CoA into mitochondria for fatty acid oxidation . IncreasGerm-free mice have a consistently raised level of phosphorylated acetyl CoA carboxylase (Acc) and carnitine palmitoyltransferase-1 (Cpt-1) activity in gastrocnemius muscles and raised AMPk in liver and skeletal tissue compared to CONV mice , 76. Thi de novo lipogenesis, very low density lipoprotein (VLDL) triglyceride export, and plasma triglyceride turnover leading to improved lipid and glucose metabolism [α/β) which help in absorption, intracellular transport, and systemic transport of bile acids into the liver by enterohepatic circulation  are ligands for the farnesoid x receptor (FXR) which plays a key role in the control of hepatictabolism . By bindculation . Study o colitis . in vitro and in vivo reduced GLP-1 secretion and impaired glucose tolerance [The gut microbiota, through production of SCFA, may affect host energy metabolism and development of obesity by changing the hormonal milieu in the intestine and other visceral organs . Glucagoolerance .SCFA including acetate, propionate, and butyrate act as ligands for the activation of G protein coupled receptors 43 and 41 (GPR41 and GPR43) which are expressed by gut epithelial cells, endocrine cells, and adipocytes. GPR43 in white adipose tissue act as sensors of postprandial energy excess and regulate energy expenditure and hence body energy homeostasis. GPR43 and GPR41 enhance insulin sensitivity and activate the sympathetic nervous system at the level of the ganglion to prevent excess energy deposition in adipose tissue and enhance energy expenditure in other tissues such as liver and muscles . GPR43 dBifidobacteria are inversely associated with the development of fat mass, glucose intolerance, and bacterial lipopolysaccharide (LPS) in the blood via SCFA-induced stimulation of PYY and ghrelin. Intervention with prebiotics such as dietary fructans or oligofructose stimulates bifidobacterial growth  and reduOverall, this evidence suggests that alteration in the gut microbiota may affect hormonal status via GLP-1 and G protein coupled receptors. These hormonal changes bring a change in satiety, food intake, and overall metabolic status of an individual that could affect host adiposity. Whether this relationship is causal needs further investigation.α, IL-6, and leptin [α, and IFN-α) which increase the risk of cardiovascular diseases [Emerging evidence suggests close ties between metabolic and immune systems . Obesityd leptin . Inflammd leptin . Intra-adiseases . These pThe gut microbiota may contribute to chronic low-grade inflammation and obesity via the absorption of bacterial LPS, an outer membrane component of Gram negative bacteria, which is increasingly recognized as a player in chronic low-grade inflammation, a hallmark of obesity. Bacteroides, Bifidobacterium species, and Eubacterium rectale-C coccoides group [ ob/ob (CD14 −/−) mice has been shown to protect against diet induced obesity and inflammation in mouse models [Cani et al. (2007) demonstrated the link between LPS and metabolic disease by infusing bacterial LPS subcutaneously into germ-free mice for 4 weeks which produced the same level of metabolic endotoxemia as by high fat diet . Furtheres group . Additioe models .Lactobacilli/Enterococci, E. rectale/C. coccoides, Bacteroides, and sulphate reducing bacteria) [Alteration in the gut microbiota is linked to changed gut barrier function  and may acteria) . GLP-1 hacteria) . Lactobacillus, Bacteroides/Prevotella, and Bifidobacteria) of obese ob/ob mice fed high fat diet [The association of high fat diet with subclinical or clinical inflammation in obesity has been investigated in several studies and there is a clear evidence to suggest that consumption of high fat diet is associated with metabolic endotoxemia and 2-3-fold increase in bacterial LPS levels in the blood . Howeverfat diet . This waIn contrast, de la Serre et al. (2010) suggested that high fat diet induced intestinal inflammation in obese Sprague-Dawley rats may cause hyperphagia and obesity by impairing the regulation of food intake. However, changes observed in the gut microbiota were independent of lean and obese phenotype . High fa ob/ob mice compared with lean mice [Cannabinoid receptors 1 and 2 (CB1 and CB2) are G proteins activated by the endocannabinoid (eCB) system. The eCB system is composed of endogenous lipids and plays an important role in adipogenesis, as studied in genetically obese mice models. Two of the most widely studied lipids in the eCB system are N-arachidonoylethanolamine and 2-arachidonoylglycerol. The level of eCB components is inversely related to obesity and type-2 diabetes as both the conditions are associated with a higher tone of eCB system. Furthermore, the expression of CB1 and CB2 degrading enzymes (fatty acid amide hydrolase) is increased in adipose tissue of obeseean mice . in vitro and in vivo studies [ ob/ob obese mice with disrupted gut barrier and metabolic endotoxemia improves gut permeability and reduces body weight, compared with lean littermates [Bacterial LPS regulates the expression of cannabinoid receptors via the LPS receptor signalling system shown in both studies . This in studies . Using Ctermates . The gut db/db mice suggested that the abundance of Gram negative bacteria, higher Firmicutes and Proteobacteria, and lower Bacteroidetes were correlated with upregulation of apelin and APJ expression. This was shown to be the result of direct action of bacterial LPS on the expression of apelin and APJ mRNA in obese diabetic mice through chronic low-grade inflammation [A study by Geurts et al. 2011) in obese leptin resistant11 in obeammation . EndocanIn summary, bacterial LPS increase the tone of eCB system and increase the expression of apelin/RPJ system in adipose tissue. However, how far gut microbiota population contribute to the actions of eCB and apelin/APJ and eCB in obesity is unknown. This has opened yet another area of interest in the role of gut microbiota in obesity. ob/ob mice, and Sprague-Dawley rats. Initial evidence suggesting a strong association of the gut microbiota with obesity was explored in a series of studies using germ-free and CONV mice. Components of gut microbiota acting as triggers in the development of obesity [The evidence from animal studies has thus far concentrated on studies which looked at the interplay of diet, gut microbiota, and metabolic changes  in animal models such as wild-type mice, leptin deficient obesity  and the  obesity  suggeste obesity , 3, 4 an ob/ob mice models [However the gut microbiota are located at the interface of environment and host. The effect of environmental factors particularly diet may therefore be highly significant and contribute to changes in the gut microbiota composition and function and ultimately their phenotype (obese or lean microbiome) . Ingestie models , and imme models  showing  Bacteroides, a relative increase in favour of Firmicutes [α) and nuclear factor-kappaB (NF-kappaB), which play a major role in promoting inflammation [Furthermore, observations from studies on GF/CONV mice and Sprague-Dawley rats suggest that a high fat diet, especially HF Western diet, is associated with increased adiposity, reduced bacterial diversity , reducedrmicutes , and higrmicutes . Moreovermicutes  such as ammation , immune ammation . This su ob/ob mice, genetically prone to obesity, indicated that although the obese phenotype is characterised by a particular set of gut microbiota, change in caloric load and diet redistributes the equilibrium that may be independent of the genotype or phenotype (obese or lean) [Studies in leptin deficientor lean) . Changesor lean) . The assor lean) .Faecal transplantation studies support the causal role of the gut microbiota in the aetiology of obesity. Transplantation of gut bacteria from obese human twins to lean mice caused not only obesity but also a higher number of genes involved in detoxification and stress response, biosynthesis of cobalamin, essential and nonessential amino acids, and gluconeogenic pathways. In contrast, animals with lean-transplanted microbiota exhibited genes capable of fermenting plant polysaccharides and producing butyrate and propionate . Additio ob/ob animal model [It is however interesting to observe that lean animals cohoused with obese cage mates are reported to develop obesity and obesity related microbiota and metabolism in some studies  but not al model , 76. Howal model . de Wit al model . High fa Ruminococcaceae and increase in Rikenellaceae with HF compared to CARB) but also the metabolome and proteome of the individual groups [n = 3) which might have contributed to variation within the groups.Several other studies suggested similar changes in gut microbiota and the presence of genes for lipid metabolism in animal models using different dietary regimens , 38, 99 l groups . AlthougIn conclusion, the relationship of gut microbiota with diet and metabolic disorders has been studied in a variety of animal models. There is controversy as to whether these changes are attributable to the diet itself or are caused by the gut microbiota. Studies in germ-free mice suggest the gut microbiota are the critical player in inflammation, development of immunity, and host metabolic regulation. However, diet is also considered a confounding factor that determines a change in gut microbiota and obesity because the diversity of gut microbiota has not been found to be different between wild-type and certain genetic models of obese mice.Discrepancies between and within studies could be attributed to the selection of animals (rats versus mice) and individual strains. A recent study by Walker et al. (2014) observed a distinct microbiome and metabolome in two strains of C57BL/6J and C57BL/6N mice . Some di Bacteroides ratio [Evidence linking the gut microbiota with obesity in humans is thus far inconclusive and controversial. This may be partly due to marked interindividual variations in the gut microbiota and metabolic activity in humans with age, diet, use of antibiotics, genetics, and other environmental factors . Apart fes ratio . BridginThe first evidence showing higher Firmicutes and lower Bacteroidetes in obese versus lean adults before the onset of dietary intervention was presented by Ley et al. (2006) , followeThe energy harvesting capability of the gut microbiota in obese subjects is thought to be set at a higher threshold than in the lean with or without differences in the relative abundance of the gut microbiota. Obese adults had higher individual and total SCFA than lean adults in the absence of any difference in the relative abundance of major gut bacterial phyla . Moreove Eubacterium rectale (a butyrate producing bacteria) to ~10% and Ruminococcus bromii (an acetate producer) to ~17% compared with ~4% in volunteers consuming nonstarch polysaccharides [ Collinsella aerofaciens, a member of Actinobacteria. This shows the substantial effect of diet on the gut microbiota and its energy harvesting capability [ Roseburia and Eubacterium rectale [On the other hand, evidence also suggests that diet plays an important role in altering the proportion of gut microbiota in individuals because the amount and type of bacteria change significantly with diet , 67. Thipability , 67. Sim rectale . Bacteroides) have been observed in children who were exposed to antibiotics in early childhood [ Bacteroides and significantly higher E. coli and Staph. aureus in overweight and obese compared to normal weight pregnant women [Long term changes in gut microbiota  may beneficially affect short term energy homeostasis in weaning infants [In addition to compositional differences between lean versus obese subjects , functio infants . However infants . Staphylococci in breast fed children at age of 6 and 12 months had a negative correlation with overweight and obesity at 7 years [n = 5914) at age of 4, 7, 15, and 23 years showed that although children born with caesarean section had ~50% higher prevalence ratio of obesity, this effect was lost when adjusted for socioeconomic, demographic, maternal, anthropometric, and behavioural factors [In this context, factors affecting colonisation of the gut microbiota in the newborn from before birth to early and late childhood might play an important role. However, the role of these factors in establishing a gut microbiota with tendency towards obesity or allergic disorders in later life is controversial. For example, higher numbers of Bifidobacteria and lower numbers of 7 years . Similar 7 years . However factors . Bacteroides in obese and nonobese subjects on different dietary regimens [Controversies exist as to whether or not obese and nonobese individuals host a particular type of bacterial phyla or enterotype and whether the response of the gut microbiota to diet differs. Correlation of BMI withregimens  is uncleregimens , adding The prevalence of obesity has increased in pandemic proportions in adults and children. Several factors have been identified to explain the aetiology and pathogenesis of obesity including diet, lifestyle, environmental factors, and host genetic factors. However, none of these fully explain the aetiology of obesity and the search for possible causes continues. The gut microbiota have been advocated as one factor affecting host energy homeostasis through several putative mechanisms investigated in mice models and human studies. However, several studies have suggested a profound effect of diet on the gut microbiota which modify host metabolism towards a lean or obese phenotype.The evidence linking gut microbiota to the increasing epidemic of obesity is too contradictory and inconclusive to prove a “cause or effect” relationship. This may be due to differences in methodology, study design, control of diet, genetic propensity of individuals to obesity, and other lifestyle factors. Moreover, faecal samples are the usual source of gut microbiota which may not represent the true picture of the colonic microbial population. Access to the full length of the gut is restricted for medical or ethical reasons. In addition, differences between animals and human beings including intestinal microbiota, metabolic rate, and length of intestine, caecal fermentation, coprophagy, and genetic variability limit the extrapolation of results from animal studies.