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BACKGROUND Schistosomiasis is a disease caused by Schistosoma. Due to its complex life cycle, evolutionary position and sexual dimorphism, schistosomes have several mechanisms of gene regulation. MicroRNAs (miRNAs) are short endogenous RNAs that regulate gene expression at the post-transcriptional level by targeting mRNA transcripts. OBJECTIVES Here, we tested 12 miRNAs and identified their putative targets using a computational approach. METHODS We performed the expression profiles of a set of miRNAs and their putative targets during the parasite’s life cycle by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). FINDINGS Our results showed differential expression patterns of the mature miRNAs sma-miR-250; sma-miR-92a; sma-miR-new_4-3p; sma-miR-new_4-5p; sma-miR-new_5-5p; sma-miR-new_12-5p; sma-miR-new_13-3p and sma-miR-new_13-5p. Interestingly, many of the putative target genes are linked to oxidative phosphorylation and are up-regulated in adult-worms, which led us to suggest that miRNAs might play important roles in the post-transcriptional regulation of genes related to energetic metabolism inversion during parasite development. It is noteworthy that the expression of sma-miR-new_13-3p exhibited a negative correlation on SmNADH:ubiquinone oxidoreductase complex I. MAIN CONCLUSIONS Our analysis revealed putative miRNA genes related to important biological processes, such as transforming growth factor beta (TGF-β) signaling, proteasome regulation, glucose and lipid metabolism, immune system evasion and transcriptional regulation.

Differences in the timing of exoskeleton melanization and sclerotization are evident when comparing eusocial and solitary bees. This cuticular maturation heterochrony may be associated with life style, considering that eusocial bees remain protected inside the nest for many days after emergence, while the solitary bees immediately start outside activities. To address this issue, we characterized gene expression using large-scale RNA sequencing (RNA-seq), and quantified cuticular hydrocarbon (CHC) through gas chromatography-mass spectrometry in comparative studies of the integument (cuticle plus its underlying epidermis) of two eusocial and a solitary bee species. In addition, we used transmission electron microscopy (TEM) for studying the developing cuticle of these and other three bee species also differing in life style. We found 13,200, 55,209 and 30,161 transcript types in the integument of the eusocialApis melliferaandFrieseomelitta varia, and the solitaryCentris analis, respectively. In general, structural cuticle proteins and chitin-related genes were upregulated in pharate-adults and newly-emerged bees whereas transcripts for odorant binding proteins, cytochrome P450 and antioxidant proteins were overrepresented in foragers. Consistent with our hypothesis, a distance correlation analysis based on the differentially expressed genes suggested delayed cuticle maturation inA. melliferain comparison to the solitary bee. However, this was not confirmed in the comparison withF. varia. The expression profiles of 27 of 119 genes displaying functional attributes related to cuticle formation/differentiation were positively correlated betweenA. melliferaandF. varia, and negatively or non-correlated withC. analis, suggesting roles in cuticular maturation heterochrony. However, we also found transcript profiles positively correlated between each one of the eusocial species andC. analis. Gene co-expression networks greatly differed between the bee species, but we identified common gene interactions exclusively between the eusocial species. Except forF. varia, the TEM analysis is consistent with cuticle development timing adapted to the social or solitary life style. In support to our hypothesis, the absolute quantities of n-alkanes and unsaturated CHCs were significantly higher in foragers than in the earlier developmental phases of the eusocial bees, but did not discriminate newly-emerged from foragers inC. analis. By highlighting differences in integument gene expression, cuticle ultrastructure, and CHC profiles between eusocial and solitary bees, our data provided insights into the process of heterochronic cuticle maturation associated to the way of life.Author SummaryFrom our previous observation that bees with distinct habits of life, eusocial and solitary, exhibit different degrees of cuticle melanization and sclerotization at the emergence, we decided to analyze the genetic signatures and ultrastructure of the integument, as well as the CHC profiles that could be involved in cuticle maturation. The expression patterns of certain genes involved in the melanization/sclerotization pathway, chitin metabolism, cuticle structure, and also regulators of cuticle renewal and tanning, in addition to other genes, might be grounded the slow process of cuticle maturation in the eusocial bees in comparison to the solitary ones. The electron micrographs revealed differences in the timing of cuticle deposition for the eusocial and solitary species. Among the identified CHCs, the proportions and quantities of n-alkanes in the developing cuticle are consistent with the faster cuticular maturation in the solitary bee, thus supporting our hypothesis.

The ACE2 (angiotensin-converting enzyme 2)/Ang-(1–7) [angiotensin-(1–7)]/MAS axis of the RAS (renin–angiotensin system) has emerged as a pathway of interest in treating both cardiovascular disorders and cancer. The MAS protein is known to bind to and be activated by Ang-(1–7); however, the mechanisms of this activation are just starting to be understood. Although there are strong biochemical data regarding the regulation and activation of the AT1R (angiotensin II type 1 receptor) and the AT2R (angiotensin II type 2 receptor), with models of how AngII (angiotensin II) binds each receptor, fewer studies have characterized MAS. In the present study, we characterize the MAS promoter and provide a potential feedback mechanism that could compensate for MAS degradation following activation by Ang-(1–7). Analysis of ENCODE data for the MAS promoter revealed potential epigenetic control by KRAB (Krüppel-associated box)/KAP-1 (KRAB-associated protein-1). A proximal promoter construct for the MAS gene was repressed by the SOX [SRY (sex-determining region on the Y chromosome) box] proteins SRY, SOX2, SOX3 and SOX14, of which SRY is known to interact with the KRAB domain. The KRAB–KAP-1 complex can be tyrosine-nitrated, causing the dissociation of the KAP-1 protein and thus a potential loss of epigenetic control. Activation of MAS can lead to an increase in nitric oxide, suggesting a feedback mechanism for MAS on its own promoter. The results of the present study provide a more complete view of MAS regulation and, for the first time, suggest biochemical outcomes for nitration of the KRAB domain.

Several proteins and different species of RNA that are produced in the nucleus are exported through the nuclear pore complexes, which require a family of conserved nuclear export receptors called exportins (XPOs). It has been reported that the XPOs (XPO1, XPO5, and XPOT) are directly involved in the transport processes of noncoding RNAs from the nucleus to the cytoplasm and/or from cytoplasm to the nucleus. All three genes are present in fungi, plants, and deuterostome metazoans. However, protostome metazoan species lack one of the three genes across evolution. In this report, we have demonstrated that all three XPO proteins are present in the parasite protostome Schistosoma mansoni. As this parasite has a complex life cycle presenting several stages in different hosts and environments, implying a differential gene regulation, we proposed a genomic analysis of XPOs to validate their annotation. The results showed the conservation of exportin family members and gene duplication events in S. mansoni. We performed quantitative RT-PCR, which revealed an upregulation of SmXPO1 in 24 h schistosomula (sixfold when compared with cercariae), and similar transcription levels were observed for SmXPO5 and SmXPOT in all the analyzed stages. These three XPO proteins have been identified for the first time in the protostome clade, which suggests a higher complexity in RNA transport in the parasite S. mansoni. Taken together, these results suggest that RNA transport by exportins might control cellular processes during cercariae, schistosomula, and adult worm development.

We evaluated the expression of the Fas receptor gene in Vero cells infected with the Lederle vaccine strain of canine distemper virus using RT-PCR. Vero cells were plated, and after being grown for 24 h in MEM with 5% FBS, 80-90% confluent monolayer cultures were infected with the virus. The cells were harvested at 3, 6, 9, and 15 h post-infection. Uninfected Vero cells were used as a control. Total RNA was isolated from Vero cells using 1 mL Trizol(®) LS, and RT was performed using 2 μg total RNA. Primer pairs for RT-PCR amplification for the canine distemper virus nucleocapsid gene, the S26 reference gene, and the Vero rFas gene were used to analyze expression in Vero cells. RT-PCR results revealed virus activity at 3, 6, 9, and 15 h in the virus-infected Vero cells. The S26 housekeeping gene was amplified in virus infected and control samples. However, expression of the cell death receptor Fas was detected in Vero cells only at 15 h post-infection. We suggest that the Lederle vaccine induces apoptosis by Fas receptor signaling, possibly through caspase-8 signaling rather than through mitochondrial signaling in the infected cells.

The presence of classical components of the renin-angiotensin system has been demonstrated in the male reproductive tract, mainly in the testes and epididymis. The objective of this study was to verify the localization of angiotensin (Ang)-(1-7) and its receptor Mas in human testis. The study included 12 men with previously proven fertility submitted to orchiectomy for prostate cancer and 20 infertile men submitted to testicular biopsy for infertility work-up, comprising a subgroup with obstructive azoospermia/normal spermatogenesis (n = 8) and another with non-obstructive azoospermia and severely impaired spermatogenesis (n = 12). Testicular tissue samples were processed by immunohistochemistry and real time polymerase chain reaction. Ang-(1-7) was strongly expressed in the interstitial compartment, mainly in Leydig cells, with similar intensity in all groups evaluated. The peptide was also detected in the seminiferous tubules, but with much less intensity compared to interstitial cells. The receptor Mas was equally distributed between interstitial and tubular compartments and was found in all layers of the normal seminiferous epithelium. However, neither Ang-(1-7) nor Mas were detected in the seminiferous tubules of samples with impaired spermatogenesis. The testicular samples of infertile men with impaired spermatogenesis (non-obstructive azoospermia) expressed Mas and ACE2 mRNA at lower concentrations (fold change = 0.06 and 0.04, respectively, P < 0.05) than samples with full spermatogenesis (obstructive azoospermia). This shows, for the first time, the immunolocalization of Ang-(1-7) and its receptor Mas in testes of fertile and infertile men, and suggests that this system may be altered when spermatogenesis is severely impaired.

The aim of the present work was to analyze c-fos response within the trigeminal nucleus caudalis (TNC) of pinealectomized rats and animals that received intraperitoneal melatonin, after intracisternal infusion of capsaicin, used to induce intracranial trigeminovascular stimulation. Experimental groups consisted of animals that received vehicle solution (saline–ethanol–Tween 80, 8:1:1, diluted 1:50) only (VEI, n = 5); animals that received capsaicin solution (200 nM) only (CAP, n = 6); animals submitted to pinealectomy (PX, n = 5); sham-operated animals (SH, n = 5); animals submitted to pinealectomy followed by capsaicin stimulation (200 nM) after 15 days (PX + CAP, n = 7); and animals that received capsaicin solution (200 nM) and intraperitoneal melatonin (10 mg/kg) (CAP + MEL, n = 5). Control rats, receiving vehicle in the cisterna magna, showed a small number of c-fos-positive cells in the TNC (layer I/II) as well as the sham-operated and pinealectomized rats, when compared to animals stimulated by capsaicin. On the other hand, pinealectomized rats, which received capsaicin, presented the highest number of c-fos-positive cells. Animals receiving capsaicin and melatonin treatment had similar expression of the vehicle group. Our data provide experimental evidence to support the role of melatonin and pineal gland in the pathophysiology of neurovascular headaches.

The renin-angiotensin system (RAS) is subject to sex-specific modulation by hormones and gene products. However, sex differences in the balance between the vasoconstrictor/proliferative ACE/ANG II/AT1 axis, and the vasodilator/antiproliferative ACE2/ANG-(1–7)/MAS axis are poorly known. Data in the rat have suggested the male-specific Y-chromosome gene Sry to contribute to balance between these two axes, but why the testis-determining gene has these functions remains unknown. A combination of in silico genetic/protein comparisons, functional luciferase assays for promoters of the human RAS, and RNA-Seq profiling in rat were used to address if regulation of Sry on the RAS is conserved in the homologous X-chromosome gene, Sox3. Both SRY and SOX3 upregulated the promoter of Angiotensinogen ( AGT) and downregulated the promoters of ACE2, AT2, and MAS, likely through overlapping mechanisms. The regulation by both SRY and SOX3 on the MAS promoter indicates a cis regulation through multiple SOX binding sites. The Renin ( REN) promoter is upregulated by SRY and downregulated by SOX3, likely through trans and cis mechanisms, respectively. Sry transcripts are found in all analyzed male rat tissues including the kidney, while Sox3 transcripts are found only in the brain and testis, suggesting that the primary tissue for renin production (kidney) can only be regulated by SRY and not SOX3. These results suggest that SRY regulation of the RAS is partially shared with its X-chromosome homolog SOX3, but SRY gained a sex-specific control in the kidney for the rate-limiting step of the RAS, potentially resulting in male-specific blood pressure regulation.

Ubiquitin-conjugating enzymes (Ub-E2) perform the second step of ubiquitination and, consequently, are essential for regulating proteolysis and for modulating protein function, interactions and trafficking. Previously, our group demonstrated the crucial role of ubiquitination and the Ubproteasome pathway during the Schistosoma mansoni life cycle. In the present investigation, we used a homology-based genome-wide bioinformatics approach to identify and molecularly characterise the Ub-E2 enzymes in S. mansoni. The putative functions were further investigated through molecular phylogenetic and expression profile analyses using cercariae, adult worms, eggs and mechanically transformed schistosomula (MTS) cultured in vitro for 3.5 h or 1 or 3 days. We identified, via in silico analysis, 17 Ub-E2 enzymes with conserved structural characteristics: the beta-sheet and the helix-2 form a central core bordered by helix-1 at one side and helix-3 and helix-4 at the other. The observed quantitative differences in the steady-state transcript levels between the cercariae and adult worms may contribute to the differential protein ubiquitination observed during the parasite ’s life cycle. This study is the first to identify and characterise the E2 ubiquitin conjugation family in S. mansoni and provides fundamental information regarding their molecular phylogenetics and developmental expression during intra-mammalian stages.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:58:48Z No. of bitstreams: 1 S0103-50531999000300013.pdf: 39242 bytes, checksum: 06a88b7acc4a4b8b89b176ba5dcfb254 (MD5) Made available in DSpace on 2013-08-22T18:58:48Z (GMT). No. of bitstreams: 1 S0103-50531999000300013.pdf: 39242 bytes, checksum: 06a88b7acc4a4b8b89b176ba5dcfb254 (MD5) Previous issue date: 1999-01-01 Made available in DSpace on 2013-09-30T19:54:51Z (GMT). No. of bitstreams: 2 S0103-50531999000300013.pdf: 39242 bytes, checksum: 06a88b7acc4a4b8b89b176ba5dcfb254 (MD5) S0103-50531999000300013.pdf.txt: 12814 bytes, checksum: 212fb90692ac0a120dde5ec7cff78c9a (MD5) Previous issue date: 1999-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T14:20:54Z No. of bitstreams: 2 S0103-50531999000300013.pdf: 39242 bytes, checksum: 06a88b7acc4a4b8b89b176ba5dcfb254 (MD5) S0103-50531999000300013.pdf.txt: 12814 bytes, checksum: 212fb90692ac0a120dde5ec7cff78c9a (MD5) Made available in DSpace on 2014-05-20T14:20:54Z (GMT). No. of bitstreams: 2 S0103-50531999000300013.pdf: 39242 bytes, checksum: 06a88b7acc4a4b8b89b176ba5dcfb254 (MD5) S0103-50531999000300013.pdf.txt: 12814 bytes, checksum: 212fb90692ac0a120dde5ec7cff78c9a (MD5) Previous issue date: 1999-01-01 O ácido canárico 1 foi isolado das folhas de Rudgea jasminoides. A substância isolada é um derivado triterpênico do tipo seco-lupano e teve sua estrutura elucidada com base nos dados espectrais, principalmente em experimentos de RMN a 1D e 2D. O sitosterol, o estigmasterol e os ácidos ursólico e oleanólico também foram isolados. The canaric acid 1 was isolated from the leaves of Rudgea jasminoides. Compound 1 is a seco-lupane derivative and its structure was established on the basis of spectral data, mainly by 1D and 2D NMR experiments. Sitosterol, stigmasterol and ursolic and oleanolic acids were also isolated. Universidade Estadual Paulista Instituto de Química Instituto de Botânica Seção de Fisiologia e Bioquímica de Plantas Universidade Estadual Paulista Instituto de Química

The digestive tract of ferrets is anatomically simple, with no caecum, ileocolic valve or external differentiation between the transition of ileum and colon. The species has a short large intestine that provides minor contributions to the digestive process. Aiming to better understand the digestibility efficiency of ferrets, the present study compared the digestibility of extruded diets with different amounts of macronutrients fed to dogs, cats and ferrets. Three formulations for cat maintenance were used (values in % of DM basis): high carbohydrate (HC; nitrogen-free extract (NFE) = 54 %, protein = 31 % and fat = 8 %); moderate carbohydrate (MC; NFE = 37 %, protein = 41 % and fat = 10 %); and low carbohydrate (LC; NFE = 19 %, protein = 46 % and fat = 23 %). Apparent total tract macronutrient digestibility was determined by the method of total collection of faeces. Results were compared by ANOVA, considering the diet and species effects and their interactions. Means were compared by the Tukey's test (P P P P P P

Hair ingested by licking during cat grooming can eventually coalesce into solid masses in cat gastrointestinal tract. It is believed that dietary fibre might reduce formation of these trichobezoars (hairballs). The effects of two insoluble fibre sources added to kibble diets were evaluated with respect to trichobezoar faecal excretion. Thirty-two cats and four diets were used in a randomised block design: a control diet without additional fibre, 10 % added sugarcane fibre, 20 % added sugarcane fibre or 10 % added cellulose. Animals were fed for 42 d and during three separate periods (days 15–17, 25–27 and 40–42), the cats were housed individually in metabolic cages and their faeces were totally collected. The faeces were evaluated and the trichobezoars were isolated and classified into small (2·1 cm). Means were evaluated by repeated measures ANOVA and contrasts (P P = 0·004) as well as a reduction in trichobezoar mass excretion (mg per cat per day;P P = 0·003), which were not present in the high sugarcane fibre group (P

The cysteine knot motifs are widely spread in several classes of peptides including those with antimicrobial functions. These motifs offer a major stability to the protein structure. Nevertheless, the antimicrobial activity is modulated by physicochemical properties. In this paper, we create a model of support vector machine to predict antimicrobial activity from sequences with similar motifs, based on physicochemical properties: net charge, ratio between hydrophobic and charged residues, average hydrophobicity and hydrophobic moment. The support vector machine model was trained with 146 antimicrobial peptides with six cysteines from the antimicrobial peptides database and an equal number of random sequences predicted as transmembrane proteins. The polynomial kernel shows the best accuracy (77.4%) on 10-fold cross validation. Testing in a blind dataset, we observe an accuracy of 83.02%. Through this model, proteins of varied size with a cysteine knot motif can be predicted with good reliability.

The search for novel antimicrobial peptides in free databases is a key element to design new antibiotics. Their amino acid physicochemical features impact into the antimicrobial peptides activities. The relationship between the amino acid physicochemical properties and the antimicrobial target might have a fuzzy behavior. This study proposes a sequence similarity and physicochemical search method followed by a fuzzy inference system to find the most appropriated antimicrobial peptides for each domain. The proposed system was tested with NCBI's NR protein data file and the obtained peptide sub dataset will be tested in vitro .

Endometriosis is an estrogen-dependent disease, causing pelvic pain and infertility. c-fos is an early transcription factor that has been reported to be related to estradiol-dependent cell proliferation. The aim of the present study was to assess the c-fos gene and protein expression in pelvic endometriotic implants in comparison to normal endometrium from infertile women. An open, prospective and controlled study included 15 infertile women with endometriosis and 19 control infertile women. Endometrial and endometriotic biopsies were performed at the follicular phase and the samples were processed for RT-PCR and immunohistochemistry. ERalpha mRNA levels were similar in the endometriotic implants/eutopic endometrium from women with endometriosis and in normal tissue (P = 0.649). The aromatase gene, however, was not expressed in the eutopic endometrium from either control or endometriosis groups, and was only expressed in 50% of endometriotic implants (P = 0.044). c-fos gene expression was higher in endometriotic implants (1.32 +/- 0.13; P = 0.011) than in eutopic endometrium from patients with endometriosis (0.97 +/- 0.11) or from the control group (0.91 +/- 0.05). In addition, immunohistochemistry showed a more abundant distribution of c-Fos in the stroma of endometriotic tissue compared to eutopic endometrium. These data suggest that c-fos may play a role in the molecular mechanisms of estrogen action on the induction, promotion or progression of endometriosis.

Developing survey strategies for threatened species is often limited by access to sufficient individuals to acquire information needed to design appropriate conservation measures. The goliath grouper (Epinephelus itajara) is a large reef fish, globally classified as critically endangered. In Brazil, fishing has been prohibited since 2002. Herein we investigated habitat use and abundance of E. itajara in Brazil drawn from a participative survey, which engaged volunteer divers in data-collection. A total of 188 reports were recorded between 2005 to 2011 throughout 13 Brazilian states. Our results revealed that habitat type influenced the relationship between total length and depth of occurrence of E. itajara. We observed a significant difference between habitats, artificial presented a nonlinear and natural an asymptotic relationship. This study underscores the importance of developing conservation measures addressing specific habitats and locations to enhance population recovery. Artificial structures (shipwrecks and manmade reefs) are critical habitats that must be considered as highly sensitive areas to E. itajara strategic conservation policies. We suggest the creation and implementation of marine protected areas, as key strategies for E. itajaraconservation, especially for artificial habitats and aggregation sites. O desenvolvimento de estratégias de pesquisa para espécies ameaçadas é frequentemente limitado pelo acesso a indivíduos suficientes para adquirir informações necessárias para elaborar medidas de conservação adequadas. Dentre essas espécies, o mero (Epinephelus itajara) é um peixe recifal de grande tamanho, classificado globalmente como criticamente ameaçado. No Brasil, a pesca é proibida desde 2002. Investigou-se o uso de hábitat e abundância de E. itajara no Brasil através de uma pesquisa participativa, que engajou mergulhadores voluntários na coleta dos dados. Um total de 188 relatos foram registrados entre 2005 a 2011 em 13 estados brasileiros. Os resultados revelaram que o tipo de hábitat influenciou a relação entre o comprimento total e profundidade de ocorrência de E. itajara. Observou-se uma diferença significativa entre hábitats: hábitat artificial apresentou uma relação não linear e hábitat natural uma relação assintótica. Este estudo ressalta a importância do desenvolvimento de medidas de conservação abordando hábitats e locais específicos visando a recuperação populacional. Estruturas artificiais (naufrágios e recifes feitos pelo homem) são hábitats críticos que devem ser considerados como áreas relevantes para as estratégias políticas conservação de E. itajara. Sugere-se a criação e implementação de áreas marinhas protegidas como uma estratégia fundamental para a conservação de E. itajara, especialmente em hábitats artificiais e locais de agregação.

The main purpose of this study was to describe a novel missense mutation (p.D179H) found in a Western Sicily family and to examine the genetic and audiologic profiles of all family members by performing a GJB2 and GJB6 mutations analysis and a complete audiologic assessment. The proband was a 3-month-old infant with a congenital profound sensorineural hearing loss; direct sequencing of the GJB2 revealed the presence of a c.35delG mutation in the heterozygous state and a heterozygous G[C transition at nucleotide 535 in trans; this novel mutation, called p.D179H, resulted in an aspartic acid to histidine change at codon 179. It was also evidenced in the heterozygous state in two members of this family, both with normal hearing. No GJB6 mutations were evidenced in all subjects studied. Considering the genotypic and phenotypic analysis of all family members, we suggest, differently from the p.D179 N mutation previously reported, a recessive mode of inheritance. Functional studies on p.D179H have to be performed to confirm our hypothesis.

Deafness is caused by a variety of facts, genetic and environmental. Regarding the acquired causes, deafness can be the consequence of prenatal infections, acoustic or cerebral trauma, and the use of ototoxic drugs. Deafness can be the only manifestation (nonsyndromic forms) or it may occur together with other phenotypic findings (syndromic forms). The majority of nonsyndromicdeafness has a genetic basis [Van Camp et al., 1997]. In recent years, deafness and hearing loss have assumed a clinical importance in the study of congenital disorders [Morton et al., 1991]. The clinical interest for hearing loss is supported by the social impact that this disorder has; if not treated, delays in the development of language and learning skills will occur [Yoshinaga-Itano et al., 1998]. In the absence of newborn screening, hearing loss might not be noticed by parents, teachers or paediatricians until the child begins to have difficulties at speaking and learning, sometimes as late as age 2 or 3 years. In genetic nonsyndromic hearing loss (NSHL) the inheritance pattern is autosomal recessive (80%), autosomal dominant (17%), X-linked (2–3%), and mitochondrial (

Point mutations and deletions of SRY gene have been described in several cases of XY gonadal dysgenesis. To date, most of these mutations affect the HMG domain of SRY which plays a central role in DNA binding activity of SRY. We report on a non-mosaic XY sex-reversed newborn girl (completely female external genitalia). The direct sequencing of SRY showed a new nonsense mutation in a codon of SRY gene flanking the 3' end of the HMG domain: a thymine is replaced by a guanine at position +387 in codon 129, resulting in the replacement of the amino acid tyrosine (TAT) by a stop codon (TAG). The new mutation of this patient provides further evidence to support the functional importance of the putative DNA binding activity of the HMG-box domain.