Clinical features included reduced consciousness, fever/hypothermia, headaches, seizures, and focal neurology. Investigation showed: raised CSF protein; pleocytosis; raised CSF:peripheral blood mononuclear cell HTLV-1 proviral load ratio; and MRI either normal or showing white matter changes in brain and cord. Four of the six previous case reports of encephalopathy in HTLV-infected patients also had HAM. Histopathology, reported in three, showed perivascular predominantly CD8+ lymphocytic infiltrates in the brain. One had cerebral demyelination, and all had cord demyelination. We have reviewed the existing six cases in the literature, together with our three new cases. In all seven with HAM, the spastic paraparesis deteriorated sub-acutely preceding encephalitis. Eight of the nine were female, and four of the seven treated with steroids improved. We propose that HTLV-associated encephalopathy may be part of the spectrum of HTLV-1-induced central nervous system disease.Human T-cell lymphotropic virus (HTLV)-1-associated myelopathy (HAM) is well described. Clinical features are predominantly consistent with cord pathology, though imaging and autopsy studies also demonstrate brain inflammation. In general, this is subclinical; however, six cases have previously been reported of encephalopathy in HTLV-1-infected patients, without alternative identified aetiology. We describe three further cases of encephalitis in the UK HAM cohort (The online version of this article (10.1007/s00415-018-8777-z) contains supplementary material, which is available to authorized users. Encephalitis refers to inflammation of brain parenchyma and should be suspected clinically in the presence of fever, headache, and altered consciousness . It may Human T-cell lymphotropic virus type 1 (HTLV-1) is a retrovirus which causes HTLV-1-associated myelopathy (HAM) in 0.25–3.7% of infected individuals . AsymptoPathological and imaging studies suggest both the brain and cord can become inflamed in HTLV-1 infection. Histopathological analyses in HAM at autopsy have demonstrated perivascular predominantly CD8+ inflammatory infiltrates throughout the central nervous system (CNS) in the presence of active-chronic spinal cord disease, compared to inactive-chronic spinal cord disease, where inflammation was noted in the spinal cord only . LeucoenThe national HTLV-1 cohort in England includes 142 patients with HAM. Three have had encephalitis—an estimated incidence of 1 per 359 person-years follow-up. This equates to a rate of 278/100,000 person-years compared with the reported annual incidence of encephalitis in England of 5.23 cases per 100,000 population [A retrospective case note review was undertaken of all patients with HAM in our UK cohort who developed encephalitis (1995–2017).The literature was reviewed using PubMed. The search enquiry was (((HTLV) OR (HTLV-1) OR (HTLV-I)) AND  OR  OR ). Included in the review are reported cases of acute- or sub-acute-onset encephalopathy in patients with HTLV-1 infection. Cases where another more likely aetiology of encephalopathy was present were excluded from the discussion. The literature search was last updated in October 2017.A 35-year-old Caucasian female had an episode of encephalitis 6 years after diagnosis of HTLV-1 infection, 2 years after onset of HAM. Over 2 months, her mobility worsened and she was admitted electively for investigation. On examination, she had low-grade fever and spastic paraparesis with hyperreflexia in all four limbs, though no meningism. She could not stand unaided, while previously had mobilised with a stick. Over the next 2 days, she became pyrexial at 39 °C, but remained haemodynamically stable. She then became drowsy and disorientated and had two generalised seizures. Despite broad-spectrum antibiotics  and IV aciclovir, her GCS fell acutely to 5 before improving, though she had ongoing confusion with visual hallucinations and severe headache.20 and mild pleocytosis (9 predominantly lymphocytes per mm3). CSF protein was raised at 2.33 g/L. No organisms were identified on extended cultures. Other investigations including MR brain were unremarkable (see Supplementary Online Table). This event predated the availability of HTLV-1 proviral load measurement. Coliform bacteria grew on urine culture (sensitive to her prescribed antibiotics).Investigation revealed mild neutrophilia with elevated C-reactive protein of 32  and alanine aminotransferase 166 (0–40 IU/L). Lumbar puncture (LP) 6 h after beginning antibiotics showed opening pressure 24 cmHFour days later, she improved clinically, becoming alert and orientated, but with no recall of recent days. Cognitive problems (amnestic and language) persisted for several months before completely resolving.Haemophilus influenzae pneumonia, treated with three courses of oral antibiotics over a month.A 52-year-old Caucasian female was wheelchair dependent at initial diagnosis of HTLV-1 infection and HAM. She had recurrent HTLV-associated uveitis and a 40 pack-year smoking history. Six years after initial diagnosis, she enrolled in a clinical trial of infliximab, however, after two doses, developed a petechial rash and severe headache so was withdrawn from the trial. The headache and rash resolved over 2 months. She then developed Five months after infliximab withdrawal, her mobility deteriorated and she could no longer transfer independently. MRI brain and cervical spine revealed cervical spondylosis but nil else of note. Eight months following infliximab withdrawal, an outpatient course of 3-day pulsed IV methylprednisolone led to a marked improvement—she could again transfer independently and straight leg raise bilaterally. The improvement was short-lived. After 1 month, she developed worsening headaches.Ten months after infliximab cessation, she developed partial left sixth and third nerve palsies with anisocoria and blurred vision. In the upper limbs, there was bilateral intention tremor and left dysdiadochokinesis. In the lower limbs, she had longstanding spasticity, but now had only a flicker of movement. She was apyrexial, haemodynamically stable and oriented.3 CSF. Protein was slightly elevated at 0.67 g/L. Normal/negative investigation results are summarised in the supplementary online table. During this episode, HTLV-1 CSF proviral load was 120 per 100 CSF cells (120%), in contrast to 17.9% in PBMCs  infection with multi-dermatomal skin lesions and encephalitis, alongside worsening myelopathy. VZV was detected on CSF PCR, with 26 white cells/mm3 were seen, and protein was 0.7 g/L. VZV was not detected in the CSF.One year later, her mobility sub-acutely deteriorated and she developed hypothermia (31–33 °C) followed by acute reduction in consciousness without new focal neurology. She had no rash. On LP, 32 white cells/mm3 CSF) in all but the penultimate episode. CSF protein ranged from 0.53 to 0.85 g/L. Investigation did not reveal another recognised cause for encephalitis (Supplementary Table). No MR abnormalities were evident during the first two episodes; however, during the third, MR imaging showed ill-defined T2-weighted pontine hyperintensities . Two episodes were preceded by 2 weeks of global aphasia and all were associated with reduced consciousness. VZV was not identified by CSF PCR though CSF showed pleocytosis ; paraneoplastic limbic encephalitis associated with a breast tumour (n = 1); progressive multifocal leukoencephalopathy (n = 6); tuberculosis (n = 1); strongyloides hyperinfection (n = 1); toxoplasmosis (n = 1); and HIV (n = 1). We also identified two case reports describing women with concurrent neuromyelitis optica (NMO) [346 articles were retrieved through the initial PubMed enquiry. Of the reported cases with both HTLV-1 infection and encephalopathy, excluded from discussion were those with co-existing: haematological malignancy n = ; paraneoca (NMO) .Six case reports described HTLV-1-infected patients with encephalopathy who had no other identified explanation for their encephalopathy (Table Eight of nine cases (including patients A–C described above) occurred in women. The mean patient age at the time of encephalopathy was 47 (range 13–73). One was < 20 years, four aged 30–50, and four aged > 50. Four were Japanese, two Afro-Caribbean, two White British, and one Brazilian. In the seven patients who also had HAM, the time between myelopathy onset and encephalopathy onset ranged from < 4 weeks to 24 years. In all patients with HAM, a sub-acute prodromal deterioration in mobility occurred. Five of nine cases had fever or hypothermia at encephalopathy onset , 21. AllSix of nine patients had a CSF pleocytosis , 19, 21,Pathological findings were reported in three cases , 18, 21.Two cases have also been reported of patients with NMO and HTLV-1 infection who developed treatment-refractory encephalopathy , 16. OneIn the UK, an underlying cause is not identified in more than a third of cases of acute encephalitis . HTLV-1 One possibility is that HTLV is not the direct causative agent of the encephalopathy, but through CNS immunomodulation and effects on the blood brain barrier , infecteThe majority of reported HTLV-associated encephalopathy has been in women. HTLV-associated myelopathy is also more common in women than in men. Furthermore, being female is a risk factor for developing a more aggressive tempo of myelopathy progression in HTLV infection . Sexual HTLV-1 infection should be considered a possible cause of encephalopathy, where no other aetiology is identified, particularly in patients from endemic areas or where there is co-existing myelopathy. HTLV-1 may also increase the risk of encephalopathy in patients with existing risk factors such as co-infections or autoimmune conditions. If HTLV-associated encephalopathy is suspected, IV corticosteroids may be an effective initial treatment option.Supplementary material 1 (PPTX 37 kb)Below is the link to the electronic supplementary material.


Children and adolescents with gender and sex diversity include (1) gender-nonconforming and transgender individuals for whom gender identity or expression are incongruent with birth-assigned sex  and (2) individuals who have differences in sex development (DSD). Although these are largely disparate groups, there is overlap in the medical expertise necessary to care for individuals with both gender and sex diversity. In addition, both groups face potential infertility or sterility as a result of desired medical and surgical therapies. The Ann & Robert H. Lurie Children's Hospital of Chicago (Lurie Children's) gender and sex development program (GSDP) provides specialized multidisciplinary care for both transgender and DSD patients. In response to patient concerns that recommended medical treatments have the potential to affect fertility, the Lurie Children's GSDP team partnered with experts from the Oncofertility Consortium at Northwestern University to expand fertility preservation options to gender and sex diverse youth. This article summarizes the results of a meeting of experts across this field at the annual Oncofertility Consortium conference with thoughts on next steps toward a unified protocol for this patient group. Specifically, the Lurie Children's GSDP team documented risks of infertility, the effects of medical and surgical intervention, and psychosocial concerns and ethical considerations in each population.Oncofertility emerged over the last decade as a field that aims to preserve fertility for cancer patients undergoing treatments, including chemotherapy and radiation, which can lead to infertility.3Although members of both the transgender and DSD communities face infertility, the reasons for fertility challenges largely differ between the two groups. Transgender youth possess inherently normal reproductive capability, but face potential infertility as a result of medical treatments intended to facilitate phenotypic transition to an affirmed gender. Pubertal suppression treatment, prescribed to youth with gender dysphoria as early as Tanner stage 2 of puberty, pauses the development of an undesired puberty, including some irreversible secondary sexual characteristics, but also prevents maturation of primary oocytes and spermatogonia to mature oocytes and sperm. Gender-affirming hormone treatment with exogenous estrogen and testosterone in adolescence may affect fertility, but the threshold at which sex steroid treatment impairs fertility is unknown.3 Furthermore, DSD conditions may be associated with elevated risk for germ cell cancer4; thus, traditionally, gonadectomy was recommended for some DSD conditions, in which infertility was presumed and risk of germ cell cancer was high.In contrast, DSD conditions are often associated with abnormal gonadal development, progressive gonadal failure over the first two decades of life, and/or abnormal hormone production, all of which can cause infertility.From these early efforts, the Lurie Children's GSDP team synthesized six representative clinical scenarios  and 2 toin vitro maturation techniques, which are currently experimental.5 While still experimental, there have been at least 60 live births following tissue transplant and either in vitro fertilization with induction of oocyte maturation or natural pregnancies.6 For those with testicular tissue, sperm is obtained for cryopreservation through masturbation or testicular sperm extraction. Testicular tissue cryopreservation is also possible, but as with ovarian tissue cryopreservation, remains investigational.7Individuals who have completed pubertal development are reproductively mature. For those with ovarian tissue, options for fertility preservation include the following: (1) embryo banking, which requires hormonal stimulation for retrieval and a suitable sperm donor, (2) oocyte banking, also requiring hormonal stimulation for retrieval, and (3) ovarian tissue cryopreservation, which requires retransplantation in the future, or Attendees discussed scenarios 1a–b noting that postpubertal transgender youths face specific challenges surrounding fertility preservation. First, given that the threshold at which gender-affirming hormones affect fertility is unknown, optimal timing for fertility preservation is likely before initiation of hormone treatment and, therefore, pursuing preservation procedures may delay hormone treatment. It can take several weeks from the time a patient expresses interest in fertility preservation to actually moving forward with scheduled procedures. In addition, an oocyte harvesting cycle, specifically, requires on average an additional 2 weeks and hormonal stimulation of the ovaries. Transgender individuals often experience long-standing gender dysphoria and may find delaying gender-affirming hormone treatment, even by a few weeks, distressing. Furthermore, among transmasculine individuals, hormonal stimulation of the ovaries results in increased estrogen, which patients may find unacceptable.8 suggesting that congruence between gamete type and gender identity is not necessary for individuals who choose to pursue genetic parenthood.Second, the techniques required for preservation are invasive and may exacerbate body dysphoria, which is frequently present among transgender individuals. Oocyte harvesting requires vaginal penetration for ultrasound monitoring and for oocyte retrieval. The simplest method of sperm retrieval is by masturbation. Such penetration or stimulation of anatomy incongruent with gender identity may be particularly distressing. Third, the harvested gametes will not match gender identity. To our knowledge, no qualitative studies exist that examine whether the type of gametes to be harvested would impact an individual's choice to pursue fertility preservation. There have been documented cases of biological parenthood in transgender individuals,9 and during which sensitivity to privacy and emotional and cognitive functioning is particularly essential.10 Adolescence also marks a developmental period during which sexuality and sexual identity are explored.11 As such, a DSD diagnosis, particularly one in which an individual's gonads are incongruent with gender identity and assigned sex at birth , may devastate an adolescent's sense of personal identity and self-esteem10 precluding careful consideration of options for fertility preservation.Review of scenarios 2 a and b elicited issues specific to postpubertal youth with DSD. As noted above, reduced inherent fertility potential is a strong possibility in some DSD conditions, thus there is no guarantee of viable oocytes or sperm. Premature gonadal failure is common; however, due to variation in timing, the window of opportunity for preservation is unpredictable. Moreover, youth and parents may be heavily burdened as they consider the complicated topic of fertility preservation while weighing the risks and benefits of potential gonadectomy to prevent germ cell cancer. Also specific to this population, many DSD conditions are known to be caused by a genetic mutation, with the possibility of transmission to offspring. As such, it is important to discuss risk for intergenerational transmission of DSD and availability of preimplantation genetic screening. As with the transgender population, gametes may not match gender identity. Anecdotally, DSD clinical care providers have expressed concern that patients and families may struggle to conceptualize the use of gametes incongruent with gender identity, but studies have not evaluated this question, thus the implications on an individual's decision to pursue fertility preservation are unknown. Finally, timing of DSD diagnosis may influence an individual's willingness to carefully evaluate options for fertility preservation. Many of these conditions are detected during adolescence, a developmental period that is critical for identity formation12In vitro follicle growth of ovarian tissue or maturation of sperm, however, remains investigational. As such, attendees unanimously agreed that such procedures must be performed under a protocol approved by an institutional review board (IRB) at an institution with expertise in these techniques. Protocols should be specific to the population, either gender or sex diverse youth. Inclusive language, such as gonad, rather than ovary or testicle, allows for preservation of either type of tissue, regardless of gender identity and is inclusive of individuals with ovotesticular DSD, who possess both ovarian and testicular tissue.Maturation of gametes occurs during puberty, as shown in 13 Pubertal suppression may alleviate psychological distress related to ongoing or anticipated pubertal changes, prevent progression of secondary sexual characteristics incongruent with affirmed gender identity, allow time for further exploration of gender identity, and provide families more time for discussion and decision-making.14 Adolescents with persistent gender dysphoria may go on to initiate later in adolescence. An additional effect of GnRHa, however, is preventing maturation of germ cells, which could be used for biological fertility potential. Preservation of ovarian or testicular tissue using IRB-approved protocol could be pursued any time before the use of gender-affirming hormones. Alternatively, the adolescent could allow some pubertal development to mature gametes, before hormone treatment, but there was consensus that this option would likely be less desirable to the youth and antithetical to the use of GnRHa for prevention of secondary sexual characteristics. The group agreed that discussion of the fertility effects of GnRHa therapy should begin before starting treatment. The gender care provider most well-known to the individual and family should initiate and repeat these discussions. Ideally, these conversations would also take place with the individual's mental health provider during the recommended assessment of readiness for medical interventions as outlined by WPATH15 and Endocrine Society clinical practice guidelines.13Considerations specific to pre- and peripubertal transgender youth with gender dysphoria include decisions about whether and when to pursue medical treatment to facilitate physical transition. Current guidelines for medical treatment of transgender youth with gender dysphoria recommend gonadotropin releasing hormone agonists (GnRHa) to pause pubertal development among early pubertal youth as early as Tanner Stages 2–3.As for postpubertal individuals with DSD, pre- and peripubertal youth with DSD may face uncertainty about inherent fertility potential and potential gonadal insufficiency. The group agreed that these questions complicate recommendations for the optimal timing of fertility preservation. Preliminary discussions may begin shortly after birth, in the case of an infant diagnosed with a DSD. Completing diagnostic testing and understanding the nature of the DSD condition can be a lengthy process of adjustment for parents, requiring patience and continued discussion. Conversation about fertility must continue in earnest if considering gonadectomy. Risk of progressive gonadal failure in early childhood, in conditions such as XO/XY Turner Syndrome and Klinefelter Syndrome, may advance the ideal timing of preservation to a younger age when primary oocytes are still present. Again, the group recognized the possibility of transmitting a genetic condition to offspring, highlighting the need to weigh this risk and consider options for preimplantation genetic screening.The group recommends a team approach for discussing and implementing fertility preservation procedures in gender and sex diverse individuals. Working group attendees offered experience with a range of institutional practice and team members. This led to an understanding that a proper fertility preservation team at one institution may be different than at another, based on the experience and availability of providers. For example, there can be overlap in roles of an endocrinologist and adolescent medicine specialist in the care of transgender youth or in surgeons' roles given varying experience with fertility preservation. In general, however, participants agreed that the team should include members with training in disciplines as shown in In addition, the group agreed that including other stakeholders will be essential in guiding the direction of this field. Members of this working group are part of the National physicians cooperative (NPC), a national interdisciplinary group of specialists, which provides fertility preservation clinical services to cancer patients, including basic scientists, allied health professionals, and physician groups . The NPC was born from needs associated with cancer patients, but now extends to incorporate other fields as fertility threats are identified. The subset of the NPC for gender and sex diverse individuals should seek input from youth affected by these conditions and their parents, as well as advocate for unique and important perspectives from those outside the medical field.in vitro maturation of cryopreserved prepubertal tissue is experimental, with no guarantee for successful future use of preserved immature gametes. Thus, the group suggested using the term “cryopotential,” rather than cryopreservation, to denote storage of immature tissue, where the potential for future use is still under development. Finally, ownership of the biological material must be established and documented before preservation. Adult individuals specify the fate of the material in the event of their death: donation to research, to another individual, or that the material be destroyed. Embryos made with a partner's gametes belong to the partner. In the pediatric population, current policies specify that in the event of death, tissue is either destroyed or donated for research.In discussing the options for fertility preservation in gender and sex diverse youth, the group raised many ethical concerns. First, as in many dilemmas in the pediatric population, issues arise regarding a patient's ability to participate meaningfully in medical decision-making . While fertility decisions ideally are made by an individual in adulthood, postponing decision-making may result in missing an optimal window during which fertility preservation is most likely to be successful. Second, the fact that parents, as the legal guardian and proxy for the youth, make decisions for their child can raise concerns, especially if there are divergent views. For example, parents' wishes may differ from those of their child or may differ between parents sharing legal decision-making rights. Third, providers may exert pressure due to their own personal biases regarding issues such as alternative options for parenting, whether to pursue fertility preservation, and the role of preimplantation genetic screening. Fourth, there is great concern about the implications of encouraging “false hope” for fertility potential. As reviewed, fertility potential in many of these conditions is unknown and Success in fertility preservation is a complicated process, dependent on multidisciplinary expertise and specific technical abilities. Thus, the group determined that fertility preservation for gender and sex diverse individuals should be offered at centers that can provide both clinical and technical expertise, with a designated team leader. In addition to the team specified in 16In discussing other potential barriers to care, participants identified geographic and financial concerns. The scarcity of fertility preservation centers limits some individuals' ability to pursue fertility preservation, particularly if travel to a center is too burdensome. Furthermore, the cost of the preservation process itself may be prohibitive. Potential fees include those for harvesting of tissue or retrieval of gametes , consultation with fertility preservation specialists, hormones to stimulate oocyte production, tissue processing fees, and shipping of gametes or tissue to a storage facility and long-term storage. Insurance coverage for such procedures is extremely limited and state specific, although the Oncofertility Consortium has worked to address this issue.The inaugural meeting of the Gender and Sex Diversity Fertility Working Group set the stage for a new field in fertility preservation. This report synthesizes the current state of the field, gaps in knowledge, and goals to address moving forward . Ultimat


Plasmodium merozoites. Merozoites invade erythrocytes using multiple binding ligands that recognise specific surface receptors. It has been suggested that adaptation of Plasmodium parasites to infect specific hosts is driven by changes in genes encoding Plasmodium erythrocyte-binding ligands (EBL) and reticulocyte-binding ligands (RBL). Homologs of both EBL and RBL, including the EBA-140 merozoite ligand, have been identified in P. falciparum and P. reichenowi, which infect humans and chimpanzees, respectively. The P. falciparum EBA-140 was shown to bind human glycophorin C, a minor erythrocyte sialoglycoprotein. Until now, the erythrocyte receptor for the P. reichenowi EBA-140 remained unknown.All symptoms of malaria are caused by the intraerythrocytic proliferation of The baculovirus expression vector system was used to obtain the recombinant EBA-140 Region II, and flow cytometry and immunoblotting methods were applied to characterise its specificity.P. reichenowi EBA-140 ligand on chimpanzee red blood cells.We showed that the chimpanzee glycophorin D is the receptor for the P. falciparum lineage. We speculate that the P. falciparum EBA-140 evolved to hijack GPC on human erythrocytes during divergence from its ape ancestor.We propose that the development of glycophorin C specificity is spurred by the  Plasmodium falciparum [Plasmodium reichenowi, infects chimpanzees [P. reichenowi isolate was genetically characterised [P. falciparum originated from P. reichenowi, most likely by a single transfer from chimpanzees (Pan troglodytes) [Pan panicus) [P. reichenowi, P. falciparum can infect chimpanzees, albeit without severe symptoms as seen in humans [Malaria causes over a half million deaths per year, chiefly among children and pregnant women in sub-Saharan Africa, and most of the cases are caused by lciparum . A closempanzees , 3. Unticterised . Due to lodytes) , 6 or evpanicus) . Howevern humans .Plasmodium species, have revealed new phylogentic species in great apes [Laverania, six Plasmodium species were confirmed, of which P. reichenowi, P. gaboni and P. billcollinsi only infect chimpanzees, whereas P. praefalciparum, P. adleri and P. blacklocki only infect gorillas. Indeed, it was proposed that great apes are natural hosts to diverse Plasmodium species, including P. falciparum previously considered as strictly human-specific [Recently, the use of molecular tools for species identification to explore the diversity of eat apes , 9, 10. specific .Plasmodium DNA from a large collection of ape fecal samples revealed the closest relative, and likely origin of human P. falciparum, is a clade of parasites found in the western gorillas [P. falciparum, in opposition to previous theories proposing chimpanzee-human transmission of P. reichenowi-related parasite. In particular, all known strains of P. falciparum circulating in humans nowadays resulted from a single cross-transmission event from gorilla to human [Sequencing of gorillas . This fito human . These dto human –15.P. falciparum adaptation to human hosts was explored by sequencing the genomes of two closely related species, P. reichenowi and P. gaboni, parasitic in chimpanzees. While it was shown that the genomes of P. falciparum and P. reichenowi are remarkably similar, striking differences were found in the genes involved in red cell invasion, which determine host specificity [Plasmodium parasites involves multiple ligands; merozoites bind to erythrocytes using proteins that belong to two families including erythrocyte-binding ligands (EBL) and reticulocyte-binding ligands (RBL) [Plasmodium adaptation to its host.The genomic basis of the cificity , 16, 17.ds (RBL) , 19. It P. falciparum EBL family recognise different human red blood cell receptors and thus enable the merozoite to gain entry through alternative invasion pathways. Four functional P. falciparum EBL proteins have been identified so far: erythrocyte-binding antigen-175 (EBA-175), erythrocyte-binding antigen-181 (EBA-181), erythrocyte-binding ligand-1 (EBL-1) and erythrocyte-binding antigen-140 (EBA-140) [Several proteins in the EBA-140) . These pP. falciparum on the human genome. These data emphasized the crucial role that EBL proteins and glycophorins play when merozoites burgle erythrocytes.Recent results from the Malaria Genomic Epidemiology Network Project  demonstrPlasmodium falciparum merozoite ligands that bind glycophorins show distinct binding behaviors, which result in different invasion pathways. The well-studied P. falciparum EBA-175 recognizes glycophorin A (GPA) [P. reichenowi [P. falciparum and P. reichenowi EBA-140 proteins are 81% identical, and the highest degree of similarity is seen within the binding region (Region II). However, the binding specificity of the P. reichenowi EBA-140 is still poorly understood. Both ligands require sialic acid for binding. Using the surface expression of human and chimpanzee EBA ligands on COS 7 cells, it was shown that the P. falciparum EBA-140 requires Neu5Ac, while its P. reichenowi counterpart requires Neu5Gc [Plasmodium species [P. falciparum, which suggests that the EBA 175-GPA interaction is probably not the sole determinant of Plasmodium host specificity [P. falciparum Rh5 RBL ligand with basigin on erythrocytes is a major determinant of host species tropism. A (GPA) –25, whil A (GPA) –28 was s A (GPA) –33, a mi A (GPA) , 34. Bot A (GPA) , but bin A (GPA) . EBA-175ichenowi , 38. The species . The hum species . HoweverP. falciparum recognises GPC on human erythrocytes [GYPC gene, which is unique among the glycophorin genes because it contains two separate translation initiation sites [P. falciparum EBA-140 [GYPC gene homologs in six Hominidae species  reveal a C to A transversion, which results in the emergence of a new start codon present only in humans [P. reichenowi-like parasites, while GPC emerged as a new receptor in humans, targeted by P. falciparum [There is a general agreement that hrocytes –33. GPC on sites , 42. Thi EBA-140 –32. Also EBA-140 , 36, 43.n humans . Thus, hlciparum .P. reichenowi binding, we used the recombinant binding region (Region II) of P. reichenowi EBA-140 obtained in baculovirus expression vector system [P. reichenowi EBA-140 is probably the homolog of human glycophorin D on chimpanzee erythrocytes.To explain the role of GPC and GPD in r system . We demoPan troglodytes) blood was freshly collected on EDTA during non-experimental clinical veterinary practice in the Warsaw Zoological Garden. The blood was drawn from immobilised chimpanzee female during a medical intervention. This sample was used to perform diagnostic tests, and the intact remainder (0.5 ml) was used in our experiments.Chimpanzee , pH 7.4 for 2 h at 4 °C with 3 × 105 native and trypsin- and chymotrypsin-  treated chimpanzee erythrocytes. The heat-denatured Region II was used as the negative binding control. The cells were washed three times with PBS and incubated for 1 h at 4 °C with rabbit serum (diluted 1:200) raised against the whole P. falciparum EBA-140 Region II [The recombinant egion II . The celegion II , dilutedP. reichenowi or P. falciparum recombinant Region II (100 μg/ml) in TBS overnight at room temperature. The bound Region II was detected with a mouse MoAb directed against the c-myc epitope . Erythrocyte GPD was detected on the blots with MoAb 2G11 [Proteins of native and enzyme-treated chimpanzee erythrocytes or human erythrocyte membranes were fractionated by SDS-PAGE using a 10% polyacrylamide gel according to the Laemmli method  and thenoAb 2G11  , stand out against that conserved background [P. falciparum but not in P. reichenowi, while EBL-1 has a substantial deletion in P. reichenowi. Similarly, of six RBLs in the P. falciparum genome, only three  have orthologues in P. reichenowi. The Rh2 locus seems to be, the most different between these two species. Notable differences can also be seen between the rif and stevor multigene families where the numbers are much lower in the human parasite [While genomes of ckground . It was parasite .var2CSA , have sum split . Thus, i alleles . ExplainP. falciparum EBA-140 [P. falciparum EBA-140 Region II does not bind to GPD.There is general agreement that GPC is the sialylated receptor on human erythrocytes for the  EBA-140 –32. SincP. reichenowi EBA-140 remained unknown. To identify it, we used a soluble, recombinant Region II obtained in insect cells. We showed previously that the P. reichenowi Region II of EBA-140 binds specifically to chimpanzee erythrocytes in a sialic acid-dependent manner [P. falciparum EBA-140 after digestion [P. reichenowi EBA-140 RII to erythrocytes after chymotrypsin treatment is detected by both flow cytometry and Western blotting experiments.Until now, the erythrocyte receptor for the t manner . We founigestion . In our P. falciparum EBA-140 ligand [P. falciparum EBA-140 ligand does not bind to human GPD [P. falciparum EBA-140 ligand lost the ability of its ancestor to bind GPD, but developed the specificity for GPC, involving its N-glycan and O-glycans.It was shown before that GPC, but not GPD plays a role in human erythrocyte invasion mediated by 0 ligand –32. Also0 ligand , 36, 43.uman GPD , due to P. reichenowi EBA-140 on chimpanzee erythrocytes. Also, these results hint that development of EBA-140 GPC specificity may have helped P. falciparum to thrive in human erythrocytes when the human and chimpanzee lineages diverged from their ancestor.Our results show that the chimpanzee GPD is the receptor for the


US are currently threatened by the spread of several species of woody plants. We studied a coastal tallgrass prairie in Texas, USA, to determine if existing woody structure increased the supply of seeds from woody plants via dispersal by birds. Specifically, we determined if (i) more seedlings of an invasive tree (Tridacia sebifera) are present surrounding a native woody plant (Myrica cerifera); (ii) wooden perches increase the quantity of seeds dispersed to a grassland; and (iii) perches alter the composition of the seed rain seasonally in prairie habitats with differing amounts of native and invasive woody vegetation, both underneath and away from artificial wooden perches. More T. sebifera seedlings were found within M. cerifera patches than in graminoid‐dominated areas. Although perches did not affect the total number of seeds, perches changed the composition of seed rain to be less dominated by grasses and forbs. Specifically, 20–30 times as many seeds of two invasive species of woody plants were found underneath perches independent of background vegetation, especially during months when seed rain was highest. These results suggest that existing woody structure in a grassland can promote further woody encroachment by enhancing seed dispersal by birds. This finding argues for management to reduce woody plant abundance before exotic plants set seeds and argues against the use of artificial perches as a restoration technique in grasslands threatened by woody species.Woody encroachment threatens prairie ecosystems globally, and thus understanding the mechanisms that facilitate woody encroachment is of critical importance. Coastal tallgrass prairies along the Gulf Coast of the  Similarly, Battaglia, Denslow, Inczauskis, and Baer . We tested the overarching hypothesis that the presence of woody vegetation would facilitate the dispersal of other woody plant species by providing perches for birds, especially at certain times of the year when seeds have set and dispersal is high.We worked in a highly endangered grassland type  the dispersal of plant seeds, especially those of woody plants that are bird dispersed, is higher under versus away from artificial wooden perches; (3) composition of seeds being dispersed differs over time between perch treatments and among habitats with different types of vegetation ; dispersal should be higher in areas already dominated by woody vegetation that birds may be actively using compared to graminoid‐dominated areas.We tested three specific hypotheses: (1) 2Schizachyrium scoparium, Andropogon geradii, Rhycospora cauduca, and Tripsacum dactyloides. Common forb species at the site include Euthamia letocephala, Helianthus angustafolius, and Liatris pynchostachya, and common woody species include the natives M. cerifera and Baccharis halimifolia, and the invasive exotic Chinese tallow, T. sebifera. Forests dominated by T. sebifera and other woody invasive species with sparse native trees, such as Celtis laevigata, are now the most abundant habitat on the property and are a common habitat type throughout the Gulf Coast . The UHCC is a ~374 ha research property previously dominated by coastal tallgrass prairie. Around 100 ha of this rare type of grassland remains, and this prairie is normally dominated by graminoids, especially T. sebifera individuals were more common throughout or around the outside perimeter of thickets of M. cerifera than in graminoid‐dominated areas, we compared the density (the number per m2) of T. sebifera individuals in paired, similarly sized M. cerifera patches and graminoid‐dominated patches in May of 2012 (n = 125 pairs of patches). We chose patches by randomly first locating one M. cerifera patch and a randomly selected graminoid‐dominated area at least 1 m outside of the M. cerifera patch. Basal area of each M. cerifera patch was calculated by measuring the radius from the center of the M. cerifera patch to 0.3 m outside of the patch; T. sebifera seedlings in graminoid‐dominated areas were counted in a patch of the same basal area. We then walked a randomly chosen distance and located the next M. cerifera patch  and repeated the process.To test hypothesis one, that To test hypothesis two, that seed dispersal is higher under woody perches, we measured how woody perches influenced the amount of seed rain (number of seeds per month) and the number of seeds in different plant functional groups every month by deploying artificial wooden perches  constructed from untreated pine. These perches are as tall as existing woody vegetation, but likely a little more robust. Paired seed traps were placed on the ground underneath an artificial perch or ~2 m away without an artificial perch. We made seed traps by securing mesh hardware cloth (1 cm) over rectangular plastic containers (29.85 cm × 19.69 cm × 12.7 cm high). Mesh was folded underneath the top edges of the seed traps; seeds fell through the mesh hardware cloth into the container, and the hardware cloth kept other animals  from getting into the traps and eating the seeds. Each pair of seed traps was located at least 4 m from the next pair, and traps were put on the soil surface.M. cerifera patches, and areas of abundant T. sebifera saplings. Five pairs of seed traps were placed in each of three replicate locations of each habitat, for a total of 180 seed traps (n = 5 replicates* perch presence or absence * three replicate locations * three vegetation types = 90 pairs of seed traps). Seed traps were deployed from June 2012 to March of 2013 to span the period during which most plants at the site were producing seeds. We removed all seeds from seed traps every month, sorted them to morphospecies, and counted the number of each morphospecies in each trap per month. We identified 23 of 33 morphospecies we observed to species by consulting keys of local vegetation , and perches did not alter rodent activity. We did, however, regularly observe birds using the perches and found bird droppings only in seed traps that were located underneath perches.T. sebifera seedlings inside versus outside M. cerifera patches, we used a paired t test. To test for differences in total number of seeds and overall species richness, we conducted repeated measures GLMs for the total number of seeds and seed species richness. To test for differences in the number of seeds of each functional group , we used repeated measures, multivariate GLMs . All possible interactions were examined because mechanistically each interaction could be important. For example, the interaction date * perch * vegetation could arise if seed rain changes under perches differently in different vegetation types. Post hoc means comparisons for differences between vegetation types were performed using Tukey's test. To determine if particular species were driving responses of functional groups to perches, for any functional group with significant effects relevant to our hypotheses (any significant main effects of or significant interactions with perch presence), we conducted follow‐up repeated measures, multivariate GLMs as described above with numbers of seeds from individual species that composed each functional group. To look at how seed composition of different plant functional groups changed over time with and without perches, we used PCA and extracted values for any principle components with eigenvalues over 1. We examined how seed composition changed graphically by plotting the average PCA values (±SE) for each date for seed composition underneath perches or not. Data were analyzed in SAS.To test for differences in the abundance of 3Tridacia sebifera seedlings were over twice as dense underneath M. cerifera thickets as in graminoid‐dominated areas of the same size . Both the abundance and species richness of seeds found in seed traps changed over time in all three types of vegetation: overall seed abundance was highest in December in all three vegetation types  and the species richness of seeds was highest in November in all three vegetation types . Myrtle‐dominated sites, however, had more different species of seeds than other sites, especially in November . When broken into plant functional groups, graminoids and herbaceous plants followed the patterns described above, but woody plants did not, with the number of seeds not differing across date or vegetation type.es Table . The effF = 10.65, p = .002) and enhanced species richness in wax myrtle vegetation , but did not affect the total number of seeds collected , or the species richness of seeds found . Moreover, the numbers of graminoid seeds and herbaceous seeds were not affected by perches in any type of vegetation . Although perches did not increase the total number of seeds delivered by birds, they did enhance the delivery of woody seeds (Hypothesis 2). Additionally, perches altered seed composition by increasing the seed rain of two invasive of woody plant species in months when seed rain was highest (Hypothesis 3). Most likely, woody structure provides perches where animals, such as birds, either drop seeds while feeding  or pass them in their feces. This positive feedback loop may be further enhanced by increased growth and survival of woody seedlings underneath woody plants, as has been shown for T. sebifera  and B. scandens (a native but invasive woody vine in grasslands). Tridacia sebifera has a substantial outer seed coating that is high in lipids and proteins , because any increase in woody species is likely to promote the further establishment of undesirable woody plants. If changing climatic conditions, like increased variability in precipitation, promote an increase in woody structure, this positive feedback loop between woody structure and increase in seed pressure due to animal dispersal may become even more important. Quick management of increases in woody plants may be especially important for grasslands that are under threat by a woody invasive species whose seeds can persist in the seed bank for long periods of time, like those of T. sebifera seeds if the woody vegetation is removed before December, when T. sebifera seeds begin to be dispersed (Figure Our results also have implications for the best seasonal timing of management activities. In particular, control of woody vegetation in Texas may be most effective in limiting dispersal of d Figure . If woodFinally, it has been suggested that artificial perches may enhance grassland restoration efforts by increasing seed pressure (Guidetti et al., None declared.CMP, AH, and SCP conceived the ideas and designed the methodology; AH collected the bulk of the data with help from CMP; CMP analyzed the data and led the writing of the manuscript. All the authors contributed critically to the drafts and gave final approval for publication.


Neuropsychiatric disorders are common forms of disability in humans. Despite recent progress in deciphering the genetics of these disorders, their phenotypic complexity continues to be a major challenge. Mendelian neuropsychiatric disorders are rare but their study has the potential to unravel novel mechanisms that are relevant to their complex counterparts.GNB5. GNB5 encodes an atypical β subunit of the heterotrimeric GTP-binding proteins (Gβ5). Gβ5 is enriched in the central nervous system where it forms constitutive complexes with members of the regulator of G protein signaling family of proteins to modulate neurotransmitter signaling that affects a number of neurobehavioral outcomes. Here, we show that the S81L mutant form of Gβ5 has significantly impaired activity in terminating responses that are elicited by dopamine.In an extended consanguineous family, we identified a novel neuropsychiatric phenotype characterized by severe speech impairment, variable expressivity of attention deficit hyperactivity disorder (ADHD), and motor delay. We identified the disease locus through linkage analysis on 15q21.2, and exome sequencing revealed a novel missense variant in Gnb5 deficiency in mice, which manifest motor deficits and hyperactivity, and highlight a critical role of Gβ5 in normal behavior as well as language and motor development in humans.We demonstrate that these deficits originate from the impaired expression of the mutant Gβ5 protein, resulting in the decreased ability to stabilize regulator of G protein signaling complexes. Our data suggest that this novel neuropsychiatric phenotype is the human equivalent of The online version of this article (doi:10.1186/s13059-016-1061-6) contains supplementary material, which is available to authorized users. It is increasingly recognized that neuropsychiatric disorders have complex etiology and that many conditions defy classical definitions based purely on phenotypic observations . While tOne large group of genes with prominent roles in neuropsychiatric disease process encodes components of neurotransmitter signaling cascades acting via G protein coupled receptors (GPCR) . NotablyThe strength of the signaling in GPCR cascades is controlled by the members of the regulator of G protein signaling (RGS) proteins, which terminate the signaling initiated by the GPCRs by accelerating the GTP hydrolysis on the Gα subunits, thereby promoting their inactivation. RGS proteins also act as signaling thresholders preventing constitutive and uncontrolled G protein signaling in the absence of GPCR activation –11. In tGNB5 and shares only ~50 % sequence similarity with classical Gβ1–4 subunits that transmit GPCR signals and are ~90 % identical to each other. Gβ5 also appears to be the only member of Gβ family that can have cellular localization other than the cell membrane, e.g. cytosolic and nuclear ..40]. S81To further characterize the effects of the mutation on the expression of Gβ5, we heterologously expressed Gβ5 constructs in HEK293T cells and determined its levels by immunoblotting. Similar to results with lymphoblastoid cells, we observed that S81L Gβ5 mutant had lower expression levels relative to wild-type protein Fig. . FurtherC. elegans results in increased locomotor activity [GNB5 is a candidate gene for ADHD in humans [The family we present in this study provides a unique opportunity to observe the phenotypic consequence of Gβ5 deficiency in humans. GNB5 knockdown in activity . Knockoun humans . ADHD isn humans . Despiten humans , 46. Intn humans .Gnb5 KO mouse, was also variably observed in patients we describe in this study. Similarly, hyperactivity, another prominent phenotype in Gnb5 KO mouse, was only present at reduced penetrance in the patients we describe with GNB5 mutation. This could be explained on the basis that these patients have partial whereas Gnb5 KO mice have complete loss of function. Expectation is that the severity may correlate with severity of Gβ5 disruption and other genetic factors may contribute to how much Gβ5 destabilization the particular mutation would cause, e.g. strength of folding machinery, or lower tone of dopamine signaling in general in unaffected subjects. This suggests that even in this Mendelian form of ADHD, modifiers may play an important role in defining the final phenotype.Motor delay, a consistent feature in Gnb5 KO mice suggested a model where increased availability of dopamine is not accompanied by a reciprocal increase in serotonin and that this imbalance may underlie the pathogenesis of hyperactivity in these mice [GNB5-related ADHD remains to be seen.Our previously published detailed analysis of the signaling perturbation in the brains of ese mice . This moese mice . The impGnb5–/– mice, deactivation of GIRK signaling after GABA binding was significantly slower than in the wildtype counterpart. Furthermore, hippocampal CA1 pyramidal neurons from Gnb5–/– mice showed altered evoked inhibitory postsynaptic currents, an important inhibitory signaling in the nervous system [GNB5-related human phenotype because the use of the GABA agonist baclofen was found to induce marked reduction in hyperactivity in Gnb5–/– mice [GNB5 mutation given the role of hippocampal declarative memory system in language development [KCNJ6, which encode GIRK2, are known to cause a severe neurocognitive phenotype in humans [GNB5 mutation [G protein–gated inwardly rectifying K+ (GIRK/Kir3) channels play an important role in synaptic plasticity and behavior , 49. We s system . These f–/– mice . In addielopment . We alson humans . Finallymutation –54. GiveGNB5 mutation in human results in an autosomal recessive neuropsychiatric disorder that is characterized by severe language delay, fine motor delay, and incompletely penetrant ADHD phenotype. It will be of interest to examine the contribution of rare variants in GNB5 in future exome/genome sequencing studies of patients with ADHD, especially those with severe speech delay that is out of proportion to their IQ.In conclusion, we suggest that


UA‐induced injury in human umbilical vein endothelial cells. The effects of UA and phloretin on cell viability, inflammation, THP‐1 monocyte adhesion, endothelial cell tube formation, GLUT9 expression and UA uptake in human umbilical vein endothelial cells were evaluated. The changes of nuclear factor‐kappa B/extracellular regulated protein kinases signalling were also analysed. Our results showed that UA reduced cell viability and tube formation, and increased inflammation and monocytes adhesion in human umbilical vein endothelial cells in a dose‐dependent manner. In contrast, phloretin significantly attenuated pro‐inflammatory factors expression and endothelial injury induced by UA. Phloretin inhibited the activation of extracellular regulated protein kinases/nuclear factor‐kappa B pathway, and reduced GLUT9 and it mediated UA uptake in human umbilical vein endothelial cells. These results indicated that phloretin attenuated UA‐induced endothelial injury via a synergic mechanism including direct anti‐inflammatory effect and lowering cellular UA uptake. Our study suggested that phloretin might be a promising therapy for hyperuricemia‐related cardiovascular diseases.Hyperuricemia is an important risk factor for cardiovascular and renal diseases. Phloretin had shown antioxidant and anti‐inflammatory properties, but its role in endothelial injury is rarely reported. In this study, we aimed to investigate the protective effect of phloretin on  Although the normal level of UA may provide antioxidant defence in the human body, abnormal high level of blood UA (hyperuricemia) is often associated with gout, metabolic syndrome, insulin resistance, hypertension, T2DM and chronic renal diseases (CKD) Increased evidences showed that some natural products such as polyphenols had beneficial effects on cardiovascular health. Phloretin belongs to the chalcone class of flavonoid compounds that abundance in apple and pear trees, which had shown various biological properties including regulation of glucose transporters, anticancer, antioxidant and anti‐inflammatory activities SLC22A12), GLUT9 (SLC2A9), ABCG2, OATs and NPTs had been previously reported SLC2A9, SLC22A12 and ABCG2 had been proved could affect renal UA excretion and blood UA levels X. laevis oocytesCellular UA uptake is a complex processes with balance between UA reabsorption and excretion In this study, we aimed to study the protective role of phloretin on UA‐induced endothelial injury in HUVECs. We evaluated the effects of phloretin on GLUT9 expression, UA uptake and endothelial cell injury in HUVECs, and explored the potential protective mechanism of phloretin on UA‐induced endothelial injury.HUVECs obtained from ScienCell Research Laboratories (USA) were cultured in supplemented EBM‐2 basal media (EndoGRO™VEGF Supplement kit. Millipore). Uric acid  was dissolved in warmed serum‐free EBM‐2 medium and filtered with 0.22 μm syringe filter unit (Millipore) as previously described SLC2A9 (GLUT9) and control siRNAs were purchased from Shanghai Genechem Co., Ltd. , and their sequences were listed in Table HUVECs were washed with PBS twice before experiment, and then UA uptake assay was performed in serum‐free EBM‐2 media containing 500 μM UA. The culture supernatant was collected at 30 and 60 min. after incubation, and the concentration of UA was measured by commercial uric acid assay kit . After experiment, cells were digested, collected and counted using haemocytometer. The UA uptake rate was determined by normalizing the UA level to cell number in each sample.HUVECs were seeded in 96‐well plate and treated with indicated conditions. After treatment, 10 μl of CCK‐8 solution  was added to each well, and the plates were incubated for 2 hrs at 37°C. The absorbance at 450 nm was measured by microplate reader . The cell viability was calculated by normalizing optical density of the experimental group to the control group.Total RNA was extracted using Trizol reagent  and reverse‐transcribed into cDNA using an iScript cDNA synthesis kit . Real‐time polymerase chain reaction  was performed on CFX96 real‐time PCR detection system  with SYBR Green . Primers used in this study were listed in Table Cells were lysed in radioimmuno‐precipitation assay (RIPA) buffer, supplemented with protease inhibitors  and phosphatase inhibitors . The protein concentration was determined by BCA Protein Assay Kit . Equal amount of protein was subjected to electrophoresis on 10% sodium dodecyl sulphate–polyacrylamide gel (SDS‐PAGE), and then transferred to polyvinylidene difluoride membranes . The membranes were blocked with 5% non‐fat milk and incubated with primary antibodies against p‐NFκB  and NFκB , GLUT9 , IL‐1β , OAT1 , URAT1  ICAM‐1 , MCP‐1 , VCAM‐1  overnight at 4°C. After washed with PBST, the PVDF membrane was incubated with horseradish peroxidase‐conjugated secondary antibody  at 37°C for 1 hr. An enhanced chemiluminescence kit (Millipore) was used for signal detection. Protein bands were quantified by NIH Image J software and normalized to the expression of β‐actin.2 incubator. The THP‐1 cells were pre‐labelled with CM‐DiI  for 5 min. at 37°C and then for an additional 15 min. at 4°C in RPMI 1640. After treatment, the THP‐1 cells were added to each well of HUVECs and co‐cultured for 2 hrs. Each well was washed three times with PBS, and then, images were randomly captured at ten regions in each well at 200 × magnification using fluorescence microscope .Human monocyte cell line THP‐1 was cultured in RPMI 1640 supplemented with 10% FBS (Gibco) and cultured at 37°C in humidified 5% CO4 cells/well). After 2 hr of incubation at 37°C, the tube‐like structure of endothelial cells was examined by inverted microscope  at 100 × magnifications. Branching points in ten random fields per well was quantified by Image‐Pro Plus software. Inhibition percentage was expressed as percentage of the control (100%). The assay was repeated three times independently.Matrigel matrix  was thawed at 4°C, then pipetted into pre‐chilled 96‐well plates (50 μl matrigel/well) and incubated at 37°C for 1 hr. After treatment, HUVECs were collected and placed on the layer of matrigel , mouse anti‐ICAM‐1 antibody (1:200), rabbit anti‐MCP‐1 antibody (1:200) overnight at 4°C followed by incubation with fluorescein isothiocyanate (FITC)‐conjugated or tetramethylrhodamine (TRITC)‐conjugated secondary antibody for 1 hr at 37°C. Nuclei were visualized by staining with DAPI. Digital images were captured by fluorescent microscope.anova test with a value of P < 0.05 considered to significant difference. Comparison between two groups of values was performed by t‐test.All data were presented as mean ± S.D., and analysed using one‐way As shown in Figure Because high dose of phloretin (>20 μM) induced obvious cytotoxicity in HUVECs Fig. A, relatiIn TNF‐α treated HUVECs, phloretin can only reduced the ICAM‐1 level, but had no influence on IL‐1β and MCP‐1 expression Fig. A‐B. BothOur results showed that GLUT9 expressed in HUVECs, which was consistent with the positive controls including HK2 cell and rat kidney Fig. A. Phlorevia multiple mechanisms such as oxidative stress, growth inhibition, epithelial mesenchymal transition (EMT) and inflammation It was well documented that there was a strong relationship between hyperuricemia and cardiovascular diseases, and high level of UA‐induced endothelial injury Extensive epidemiologic studies suggest that elevated blood UA level is highly associated with diabetes, cardiovascular and renal diseases via directly recruiting inflammatory cells such as neutrophils, monocytes and macrophages via activating NF‐κB/ERK pathway. Phloretin is a natural polyphenolic compound derived from rosaceae family plants, which had shown antioxidant and anti‐inflammatory properties Inflammation plays a key role in the initiation and progression of vascular injury response to stress stimulation, and the release of inflammatory factors further aggravated endothelial injury SLC2A9) had been recognized as a high capacity UA transporter in human and rodents xenopus laevis oocyte model at a dose of 1 mM via inhibiting GLUT9‐mediated UA transport in HUVECs. Furthermore, we used siRNA to knockdown GLUT9 and assayed the effect of GLUT9 inhibition on inflammation in HUVECs. Our results showed that GLUT9 siRNA significantly decreased inflammatory factors expression and NF‐κB/ERK activation induced by UA, which was similar to the effect of phloretin on UA‐treated HUVECs. These results suggested that inhibition of GLUT9‐mediated UA transport also contributed to the protective effect of phloretin on UA‐induced endothelial injury.Although the anti‐inflammatory effect of the phloretin has been reported, few studies investigated the role of phloretin on cellular UA uptake and UA transporters. UA transporters such as OATs, URAT1 and GLUT9 played a key role in maintaining UA homoeostasis in various types of cells and tissues Taken together, hyperuricemia is strongly associated with endothelial dysfunction and development of diabetes, cardiovascular and renal diseases. Phloretin significantly attenuates UA‐induced vascular injury due to its anti‐inflammatory and lowering UA action. In addition, phloretin is a natural compound abundant in plant trees, and its therapeutic efficacy can be further improved by chemical or biological modification. Therefore, phloretin provides a novel and cost‐effective approach to prevent hyperuricemia‐associated vascular complications.via activation of inflammatory signalling in HUVECs, whereas phloretin significantly attenuated inflammation and endothelial injury induced by UA. We further revealed that phloretin decreased UA‐induced HUVECs injury via a synergic mechanism including direct anti‐inflammatory effect and blocking of GLUT9‐mediated UA uptake. This study suggested that phloretin might be a promising therapy to hyperuricemia‐related cardiovascular diseases.In conclusion, our study demonstrated that high level of UA‐induced endothelial dysfunction The authors confirm that there are no conflict of interests.Table S1 The sense and antisense sequences for siRNA in this study.Table S2 Primers sequences for real‐time PCR in this study.Click here for additional data file.


GnRH is secreted in short pulses and GnRH effects on its target cells are dependent upon the dynamics of these pulses. Here we overview GnRH receptors and their signaling network, placing emphasis on pulsatile signaling, and how mechanistic mathematical models and an information theoretic approach have helped further this field.Gonadotropin-releasing hormone (GnRH) acts via G-protein coupled receptors on pituitary gonadotropes to control reproduction. These are G •We describe a mechanistic model for GnRH action via its Gq/11-coupled receptors.•GnRH is secreted is pulses which can increase signaling efficiency and robustness.•Maximal responses occur at sub-maximal GnRH pulse frequency.•This may reflect incoherent feedforward loops but not upstream negative feedback.•Single cell measures reveal noisy signaling systems with marked information loss. It acts via receptors (GnRHR) on pituitary gonadotropes to control synthesis and secretion of the two gonadotropin hormones (LH and FSH) that in turn regulate gametogenesis and steroidogenesis in the gonads. LH and FSH are heterodimeric proteins with distinct β-subunits (LHβ and FSHβ) and a common α-gonadotropin subunit (αGSU) that are packaged into vesicles for release from gonadotropes. Acutely, GnRH regulates the exocytotic fusion of these vesicles with the plasma membrane whereas chronically it increases synthesis of gonadotropins and thereby controls vesicle content. There are three distinct forms of the hormone termed GnRH-I , GnRH-II and GnRH-III. The cloned GnRHR, which are members of the rhodopsin-like GPCR family, have been classified into three groups based on sequence homology. All of the cloned mammalian GnRHR are in groups I or II, and the type I GnRHR of humans, rats, mice, pigs, sheep, and horses share >80% amino acid sequence homology . Some prq, which activates PLC to generate IP3 and DAG by cleavage of phosphatidylinositol -bisphosphate , calmodulin-dependent protein kinases, the calmodulin dependent phosphatase calcineurin (Cn) and the Ca2+ dependent transcription factor NFAT  over the known system topologies. Instead, several groups have developed mathematical models for modules or pathways within the network, notably by modeling receptor trafficking, Cativation . We haveecretion A. To exp cascade . As show cascade B and C,  cascade . More re cascade . For exa cascade  is a usereversal .Fig. 2Siin vivo . To exp−7 M GnRH whereas physiologically GnRH pulses are in the low nM range  and nM range  so the d−7 M GnRH as a constant stimulus or in pulses , setting the internalization and recycling rates at 1× (again as multiples of the estimates obtained from data) or varying them by serial halving or doubling (i.e. from 0.03125× to 32×). Using the integrated PLC response as a readout we found, as expected, that with either paradigm increasing the rate of internalization reduced the response whereas increasing the rate of recycling increased it. With constant stimulation the system shows comparable sensitivity to internalization and recycling because they are equally important determinants of cell surface receptor number at equilibrium, and this is evidenced by the near symmetrical curves for internalization or recycling versus PLC activity in Extending the modeling outlined above, we simulated responses to 102+ responses with 5 min pulses of 10−7 M GnRH at varied period and at varied GnRHR internalization rates . System output was calculated as the AUC for the Ca2+ concentration over 16 h and the condition giving the highest AUC is plotted in red . As shown, the system output was greatest at 15 min period with 1× internalization, at 30 min period with 8× internalization and at 60 min period with 16× internalization. Simulations with a broader range of pulse frequencies and internalization rates -2 -2 , inductialing)-2 , down-reeceptors , and ERKeceptors . Howevereceptors  and thiseceptors , as welleceptors  but it ieceptors  and the eceptors .Fig. 5MaTaken together, the work outlined above shows how upstream negative feedback could theoretically generate bell-shaped frequency response relationships but also suggest that such feedback is insufficient to shape GnRH signaling with physiologically relevant pulsatile stimulation. Where signaling inputs to the nucleus show increasing monotonic frequency-response relationships, the obvious possibility is that feedback and/or feed-forward regulatory loops within the nucleus underlie the observed bell-shaped frequency-response relationships for gene expression. This has been explored most extensively for the FSHβ promoter, for which a number of incoherent feed-forward loops have been described. These are signaling modules that fan out from an upstream node and re-converge at a downstream node and for which the two divergent branches have different overall signs (i.e. positive and negative effects). Thus, for example, stimulation of FSHβ gene expression by GnRH is, in part, mediated by its ability to phosphorylate and activate the transcription factor CREB, but GnRH can also increase expression of the inducible cAMP early repressor (ICER), which inhibits the effect of CREB, providing both positive and negative inputs to the promoter . As note−7 M GnRH at varied period and with varied GnRHR internalization rates. 2+ and nuclear NFAT as well as predicted transcriptional responses driven by ERK or NFAT alone (ERK-DT and NFAT-DT) and in each case maximal system outputs are predicted at maximal pulse frequency. However, simulations assuming co-operative convergence of the two transcription factors at a promoter reveals non-monotonic frequency–response relationships at all three internalization rates (i.e. non-monotonic relationships due to co-operative convergence at the transcriptome rather than due to negative feedback). Interestingly, when the same parameters were used to explore GnRH concentration-dependence  the simulations suggest that GnRHR internalization influences the balance of signaling via ERK and NFAT  act at separate sites on a common gene promoter and considered three different logic gates; an “and-gate”, an “or-gate” or a “co-operative gate”. This model predicted bell-shaped frequency-response relationships when two transcription factors act co-operatively. The characteristic feature of maximal response at sub-maximal frequency was never seen with the and-gate or with the or-gate, and this behavior was predicted without negative feedback . More re52+ concentration, gonadotropin secretion, effector activation and gene expression  in light of it. The simulations outlined above effectively model the behavior of a typical GnRH-stimulated cell as representative of the whole population and ignore the cell-to-cell variation that has already been documented for GnRH effects on cytoplasmic Capression .Information theory was developed to analyze electronic communication but is now also being used to measure how reliably biological signaling systems transfer environmental information . Here, ‘The value of this approach can be illustrated by considering a simple signaling network that bifurcates and adapts over time as shown in 28), of which <1 Bit of information was transferred. Second, population-averaged measures consistently show responses to GnRH being graded over a wide range of GnRH concentrations, yet an MI of <1 implies that single cells cannot unambiguously distinguish between just two inputs (i.e. with and without GnRH). This was not due to use of a heterologous expression system because information transfer values were similar in HeLa cells (with exogenous GnRHR) and LβT2 gonadotropes (with endogenous GnRHR). It was also not restricted to the ERK pathway because information transfer from GnRHR to NFAT was <0.5 Bits in both cell models  upstream negative feedback b) co-operative convergence of distinct pathways and c) the existence of incoherent feedforward loops. Our mathematical modeling argues against (a) as it requires strong negative feedback and associated pronounced desensitization that is not evident with our pulsatile stimulation paradigms . Indeed,


The Sonic Hedgehog (Shh) signaling pathway is active during embryonic development in metazoans, and provides instructional cues necessary for proper tissue patterning. The pathway signal transducing component, Smoothened (Smo), is a G protein-coupled receptor (GPCR) that has been demonstrated to signal through at least two effector routes. The first is a G protein–independent canonical route that signals to Gli transcriptional effectors to establish transcriptional programs specifying cell fate during early embryonic development. The second, commonly referred to as the noncanonical Smo signal, induces rapid, transcription-independent responses that are essential for establishing and maintaining distinct cell behaviors during development. Herein, we discuss contributions of this noncanonical route during embryonic development. We also highlight important open questions regarding noncanonical Smo signal route selection during development, and consider implications of noncanonical signal corruption in disease. During embryogenesis, the Sonic Hedgehog (Shh) signal transduction network instructs cell fate decisions to drive tissue patterning. Corruption of Shh pathway regulation leads to developmental disorders including holoprosencephaly (HPE), Gorlin’s Syndrome, Greig cephalopolysyndactyly, and Pallister-Hall syndrome, and in some cases results in early embryonic lethality . As suchVertebrates encode three Hh family ligands: Shh, Desert Hh, and Indian Hh, with Shh being expressed most broadly . All Hh Gli1, which functions as a feed-forward activator to sustain or amplify target gene expression . In. In49]. Recent work suggests that, in addition to affecting axon guidance in the central nervous system, noncanonical Smo signaling is also utilized in the enteric nervous system to repel enteric axons of the gut. In this context, Smo signaling through the Gαi family member Gαz repels axons to prevent them from projecting into intestinal villi . Gαz is 2+ increase that triggers protein kinase Cα and integrin-linked kinase activation [2+ concentration through Gαi [As axons travel toward their final targets, neighboring axons bundle as they grow, allowing them to track along paths determined by earlier pioneer axons . Althougtivation . The docough Gαi  suggestsXenopus support that noncanonical Smo-Gαi signaling regulates post-mitotic neuronal differentiation through controlling neurotransmitter expression [2+ spikes in response to specific extracellular signals [Xenopus spinal cords has been observed to induce phospholipase C-mediated inositol triphosphate (IP3) production at the primary cilium, leading to the release of Ca2+ from internal stores and influx through transient receptor potential cation channel 1 (TRCP1). These calcium transients resulted in the establishment of a GABAergic neuronal phenotype, indicating a role for Shh signaling in post-mitotic neuron differentiation. Importantly, in this study, Shh-regulated Ca2+ modulation was found to be inhibited by PTX or constitutively active PKA, indicating that Smo-Gαi signaling is likely required [Studies in pression . During  signals ,63,64. Srequired .2+ by opening TRP Ca2+ channels in non-neuronal cell types [2+ can also inhibit activity of ciliary adenylyl cyclases 5 and 6, leading to decreased cAMP and PKA activity, and increased Gli transcriptional activity. The precise mechanism by which Smo signals to control Ca2+ in this cellular context was not determined, but was demonstrated to be PTX-insensitive, suggesting Gαi independence [2+ modulation occurs through Smo-Gαz coupling or through a novel noncanonical Smo signal.Subsequent studies in mouse fibroblasts demonstrated the ability of Smo signaling to control ciliary Call types . The spell types . In fibrpendence . HoweverThe ability of Shh to induce proliferative responses by activating mitogenic transcriptional programs is well established . Recent Drosophila [Drosophila identified the Hh pathway as the top-scoring signaling cascade capable of modulating adipocyte fate determination [An increasing body of work connects canonical and noncanonical Shh signaling with metabolism in both vertebrate and invertebrate systems . Althougosophila . In thisosophila . In a sumination . This stmination . Howevermination ,72, raismination . As such2+ channels, leading to phosphorylation-dependent activation of AMPK [An aspect of metabolism that is more clearly linked to Shh-Smo-Gαi signaling is the rapid induction of Warburg-like metabolism in adipocytes . This me of AMPK . ImportaThe ability of Smo to signal through at least two distinct effector routes provides Shh signaling the opportunity to integrate with additional signaling pathways and impact wide-ranging biological responses during development. Although knowledge of these novel signaling connections is growing, a number of open questions regarding noncanonical Smo signaling have yet to be addressed. Key among them is identification of the molecular mechanisms controlling Smo effector route selection. As discussed above, studies in pre-adipocytes suggest that Smo effector route selection can be modulated through differential small molecule binding . As suchAn additional mechanism by which signal bias might be controlled is through post-translational modification of Smo. We recently identified that loss of Smo N-linked glycosylation can shift signaling away from Gαi and toward Gli . We did It is well established in the literature that Smo sub-cellular localization controls its activity. In order to signal to Gli, Smo must traffic to the tip of the primary cilium . ConversContinued analysis of noncanonical Smo signaling is essential for a better understanding of the physiological role of Gli-independent Smo signaling during development. Going forward, the impact of noncanonical Smo signaling to disease will also need to be considered. Aberrant Shh signaling is causative in medulloblastoma and basal cell carcinoma, and is commonly activated in a number of additional tumor types . Consequ


In rare instances, recipients of organ transplants from human T-lymphotropic virus type I- (HTLV-I-) positive donors reportedly developed neurologic symptoms due to HTLV-I-associated myelopathy (HAM). We present herein two cases of HAM associated with renal transplantation from HTLV-I seropositive living-donors. The first patient was a 42-year-old woman with chronic renal failure for twelve years and seronegative for HTLV-I. She underwent renal transplantation with her HTLV-I seropositive mother as the donor, and she developed HAM three years after the transplantation. The second patient was a 65-year-old man who had been suffering from diabetic nephropathy. He was seronegative for HTLV-I and underwent renal transplantation one year previously, with his HTLV-I seropositive wife as the donor. He developed HAM eight months after renal transplantation. Both cases showed neurological improvements after the immunomodulating therapies. We tried to shed some light on the understanding of immunological mechanisms of transplantation-associated HAM, focusing on therapeutic strategies based on the immunopathogenesis of the condition. Human T-lymphotropic virus type I (HTLV-I) is a retrovirus and an etiologic agent for adult T-cell leukaemia/lymphoma (ATL)  and HTLVCase  1. A 42-year-old woman with chronic renal failure had been undergoing hemodialysis for 12 years and was seronegative for HTLV-I. She underwent renal transplantation 5 years previously, with her HTLV-I seropositive mother as the donor, but her renal function failed 11 months owing to toxaemia during pregnancy. Three years after the transplantation, she began experiencing difficulty with walking and attended our hospital. Neurological examinations demonstrated mild hyperreflexia in all extremities, especially the lower limbs. Spasticity of the lower limbs was evident, and the pathological reflexes were easily elicited bilaterally; she showed unstable and spastic walking. The cranial nerves and cerebellar systems were intact. She experienced mild sensory disturbance in all extremities. Her serum anti-HTLV-1 antibody titer was highly elevated (×1024); and Western blotting identified HTLV-I gP46, P19, P24, and P56 cerebrospinal fluid (CSF) antibodies. The CSF protein content was 42 mg/dL with a cell count of 14/mm3. Cranial magnetic resonance imaging (MRI) showed small subcortical lesions  improving from 8 to 5 after the treatment. After the treatment, repeated MRI examinations of the spinal cord were performed, and signal alterations of the atrophic spinal cord segment still remained unchanged. These observations may indicate gliosis in the spinal cord. lesions , and MRI lesions . In addiCase  2. The second patient was a 65-year-old man who had been suffering from diabetic nephropathy. He was seronegative for HTLV-I and underwent renal transplantation one year previously, with his HTLV-I seropositive wife as the donor. Eight months after renal transplantation, he presented with weakness of legs. This progressed rapidly, leaving him wheelchair-bound 12 days later. Despite receiving pulsed steroid therapy (1 g of methylprednisolone for three days), he was unable to regain his leg muscle strength, and he was therefore referred to our hospital. Neurological examinations showed flaccid paraparesis of the legs. Deep tendon reflexes were diminished, and the pathological reflexes were easily elicited bilaterally. Nerve conduction studies did not indicate any evidence of peripheral neuropathy. His serum anti-HTLV-I antibody titer was elevated (×256). The CSF protein content was 77 mg/dL with a cell count of 11 cells/mm3. Western blotting identified HTLV-I gP46, P19, P24, and P56 CSF antibodies. MRI of the thoracic spinal cord showed T2 high signal intensities with slight cord swelling . Following this, he was able to move his legs more easily and stand up with assistance. MRI examinations showed spinal cord atrophy  may be correlated with the risk of onset, but this remains unconfirmed and further studies are required. The class I allele HLA-A2 has been reported to have a preventive effect against development of HAM in patients infected with HTLV-I. However, the latency period for Case 2 who possessed the A2 allele was 8 months, whereas it was 36 months for Case 1, who did not possess the allele. These results suggested that the onset of HAM after renal transplantation in our cases is not delayed by this potential immunologic defense mechanism.There have been reports of causative correlations between transmission of HTLV-I from a carrier donor and the development of HAM in the recipient. In these cases, the use of immunosuppressants and the HTLV-I viral load have both been suggested as important for HAM development. It is thought that key roles in the pathogenic mechanisms of HAM are played by activated T cells, an exaggerated cytotoxic T lymphocyte response to HTLV-I, and induced cytokines via the activate lymphocytes; these could ultimately induce the spinal cord damage. One therapeutic strategy for HAM should therefore be to administer immunosuppressive agents. In our cases, tacrolimus and mycophenolate mofetil (MM) were used as immunosuppressants for renal transplantation. Tacrolimus binds to calcineurin and inhibits the expression of cytokine and proliferation of cytotoxic T cells. MM is an inhibitor of the type 2 isoform of inosine monophosphate dehydrogenase expressed in activated T and B lymphocytes and inhibits the proliferation of both T and B cells. Thus, both drugs could theoretically inhibit the development of HAM. However, in our cases, the use of immunosuppressants could not have significantly affected the development of HAM, given that Case 2 developed HAM within 8 months, a short latency period while taking immunosuppressive drugs. An intramuscular interferon-In conclusion, reports of HAM onset following living-donor renal transplantation are rare, and further analysis of the pathological mechanisms and treatment strategy in this neurological disorder is needed.


Protein glutathionylation is a redox-mediated posttranslational modification that regulates the function of target proteins by conjugating glutathione with a cysteine thiol group on the target proteins. Protein glutathionylation has several biological functions such as regulation of metabolic pathways, calcium homeostasis, signal transduction, remodeling of cytoskeleton, inflammation, and protein folding. However, the exact role and mechanism of glutathionylation during irreversible oxidative stress has not been completely defined. Irreversible oxidative damage is implicated in a number of neurological disorders. Here, we discuss and highlight the most recent findings and several evidences for the association of glutathionylation with neurodegenerative diseases and the role of glutathionylation of specific proteins in the pathogenesis of neurodegenerative diseases. Understanding the important role of glutathionylation in the pathogenesis of neurodegenerative diseases may provide insights into novel therapeutic interventions. Most ofin vivo . GSH canin vivo . GSH is in vivo , 6. The in vivo , 8.GSH is synthesized in the cytoplasm; it is translocated to different organelle-specific functions. It may be implicated in organelle-specific functions related to the regulation of redox homeostasis. The redox state of GSH in the endoplasmic reticulum (ER) possesses more oxidative than that in cytosol for maintaining and promoting the folding and posttranslational modifications of the proteins. GSSG is the source of oxidizing power that ensures the functional conformation of native polypeptides by the formation of required intramolecular disulfide linkages in the ER , 10. MitThe major functions of GSH are acting as the indicator of the cellular redox state, antioxidant defense, and the storage and transport of cysteine , 17. TheUnder normal physiological conditions and oxidative stress, many redox modifications can occur. Especially, cysteine thiols (-SH) of proteins can become targets of reactive oxygen species (ROS) and modulators including glutaredoxins and glutathione transferases. It can cause a chain of modifications in their oxidation state, which can conclude the fate of particular proteins. Redox modifications can enable the cysteine thiols of protein to maintain and tolerate the irreversible and reversible posttranslational modifications during oxidative stress.Protein palmitoylation is an important reversible lipid modification in which one or more cysteine thiols on a substrate are modified to form a thioester with a palmitoyl group . The revα-ketoglutarate dehydrogenase complex (KGDHC), which function in the glycolytic pathway and Krebs cycle, respectively, are regulated by glutathionylation. GAPDH is inactivated via glutathionylation on Cys149 in endothelial cells following exposure to oxidative stress [α-induced apoptosis through glutathionylation of caspase-3 [κB) is a transcription factor that upregulates the expression of many genes involved in inflammation, cell proliferation, and defense against apoptosis [κB can itself be directly glutathionylated. The glutathionylation of NF-κB subunit p50 at Cys62, which lies in the DNA-binding domain, has been shown to induce the reversible blocking its DNA-binding activity [κB kinase (IKK) β subunit is a target of glutathionylation during oxidative stress. Glutathionylation of IKKβ is reversed by Grx, which rescues the activity of kinase [Protein glutathionylation is considered as a defense mechanism to protect proteins from oxidative states that leads to irreversible damage and is the reversible posttranslational modification on cysteine thiol groups of the protein through the disulfide bond with GSH . Glutathe stress , 32. Mite stress . In addiaspase-3 . For exaaspase-3 –38. Furtpoptosis . NF-κB cactivity . Anotherf kinase . Moreovef kinase . It is af kinase , 44. Celf kinase . In contf kinase . In addif kinase . Cytoskef kinase .Thus, as discussed above, glutathionylation possesses several biological functions such as regulation of metabolism pathways, calcium homeostasis, modulation of cell signaling, apoptotic pathways, remodeling of cytoskeletal organization, inflammation, and protein folding. However, the exact role and mechanism of glutathionylation during oxidative stress have not been completely defined. Although significant progress has been made in deciphering the biological roles of protein glutathionylation in cells, several critical questions regarding the selection of target and the tight regulation of glutathionylation remain.Neurodegenerative diseases involve some common pathological and pathogenesis characteristics such as progression of neuron loss, aggregation of misfolded proteins, mitochondrial dysfunction, increased removal of iron, and neuronal cell death due to the overexposure to reactive nitrogen species (RNS) and reactive oxygen species (ROS) –51. The β (Aβ) peptide aggregation surrounded by dying neurites called senile plaques [β is associated with mitochondrial dysfunction, increased calcium levels, and breakdown of the membranes. AD follows after the accumulation of Aβ induced by oxidative stress [α-Enolase is also glutathionylated in AD brain and that this is associated with a decreased activity [Neurodegenerative diseases exhibit common pathological and pathogenesis characteristics such as the progression of neuron loss. AD is a progressive neurodegenerative disease in which there is a decrease in memory and recognition ability. There are some specific characteristics in AD pathology, including the formation of intracellular neurofibrillary tangles that result in accumulation of hyperphosphorylated tau proteins, disruption of axonal transport, and amyloid- plaques , 56. Aβ e stress . Specifiactivity . Furtheractivity , and theactivity . Tau proactivity , 62. Gluactivity , 64. Moractivity . AD braiactivity , 67. As substantia nigra of the midbrain [PARK2 (Parkin), SNCA (α-synuclein), PINK1 (Pink1), and PARK7 (DJ-1) [α-synuclein act as a core for the association with other proteins. These deposits, which consist of an ubiquitin-proteasome system and intraneuronal inclusions, are called Lewy plaques. α-Synuclein is localized in the synaptic vesicles and on the cell membranes of nervous tissue [α-synuclein including phosphorylation and nitrosylation can cause misfolding and deposition of the protein [in vivo. Furthermore, they found that DJ-1 is susceptible to glutathionylation in vitro and in vivo [in vitro studies, one can suggest from these observations that Grx may serve to deglutathionylate the subunit of mitochondrial complex I and play an important role in maintaining mitochondrial complex I activity in sporadic PD. In mice treated with MPTP to simulate PD, glutathionylation of mitochondrial NADP+-dependent isocitrate dehydrogenase has been reported [Drosophila PD model induced by loss of parkin gene, glutathionylation levels of ATP synthase β subunit are decreased and restored by expressing GST omega (GstO) [β subunit by GstO regulates mitochondria F1F0-ATP synthase activity and is important for restoration of mitochondrial function in Drosophila PD model.PD is a common neurodegenerative disorder, which occurs due to the loss of dopaminergic neurons in the midbrain . PD is cmidbrain . The fam7 (DJ-1) –72. Thes7 (DJ-1) –77. Alth7 (DJ-1) . The pros tissue . Posttra protein . DJ-1 fu protein . Recentl in vivo . These r in vivo . In addi in vivo . Despitereported . In Drosa (GstO) . GlutathALS, also known as the Lou Gehrig's disease, is a devastating adult-onset neurodegenerative disease characterized by progressive degeneration of the motor neurons. ALS leads to gradual muscle weakness, eventually leading to fatal muscle atrophy and paralysis, and death within 3~5 years of disease onset . ALS can2+-permeable transient receptor potential cation (TRPC) channel member 5 (TRPC5) in the striatum of transgenic mice and patient with HD. In vitro studies further demonstrated that TRPC5 is glutathionylated at Cys176 and Cys178 by oxidative stress such as GSH loss. This results inactivation of the calcium channel, leading to an influx of calcium into the cells and eventually death of neurons [HD is a neurodegenerative disorder characterized by loss of motor control and recognition ability. HD is caused by abnormal CAG triplet expansions, which encodes a poly-glutamine repeat at the N-terminus of the huntingtin (HTT) protein . An abno neurons . TRPC5 gProtein glutathionylation is one of the important posttranslational modifications that regulate various cellular processes by regulating protein functions and prevents irreversible oxidation of cysteine thiol in the target proteins. Numerous pieces of evidence suggest that glutathionylation of specific proteins is associated with a number of neurodegenerative diseases, including AD, PD, ALS, and HD. Protein glutathionylation can occur in response to oxidative damage and other stress and may contribute to neurodegeneration by dysregulating several cellular processes . RecentlSince protein glutathionylation is likely to regulate protein functions and cellular signaling pathways that play a critical role in pathogenesis of neurodegenerative diseases, it is an important therapeutic target for drug development. However, no therapeutic drugs are available for modulating protein glutathionylation in the progression of neurodegenerative diseases. The potential involvement of glutathionylation in human neurodegenerative diseases highlights the need of a better understanding of how diseases are caused by glutathionylation of specific proteins and if they are targets for prevention of neurodegenerative disease.