Deciphering Tacrolimus-Induced Toxicity in Pancreatic ß Cells.

ß Cell transcription factors such as forkhead box protein O1 (FoxO1), v-maf musculoaponeurotic fibrosarcoma oncogene homolog A (MafA), pancreatic and duodenal homeobox 1, and neuronal differentiation 1, are dysfunctional in type 2 diabetes mellitus (T2DM). Posttransplant diabetes mellitus resembles T2DM and reflects interaction between pretransplant insulin resistance and immunosuppressants, mainly calcineurin inhibitors (CNIs). We evaluated the effect of tacrolimus (TAC), cyclosporine A (CsA), and metabolic stressors (glucose plus palmitate) on insulinoma ß cells in vitro and in pancreata of obese and lean Zucker rats. Cells were cultured for 5 days with 100 µM palmitate and 22 mM glucose; CsA (250 ng/mL) or TAC (15 ng/mL) were added in the last 48 h. Glucose plus palmitate increased nuclear FoxO1 and decreased nuclear MafA. TAC in addition to glucose plus palmitate magnified these changes in nuclear factors, whereas CsA did not. In addition to glucose plus palmitate, both drugs reduced insulin content, and TAC also affected insulin secretion. TAC withdrawal or conversion to CsA restored these changes. Similar results were observed in pancreata of obese animals on CNIs. TAC and CsA, in addition to glucose plus palmitate, induced comparable inhibition of calcineurin and nuclear factor of activated T cells (NFAT); therefore, TAC potentiates glucolipotoxicity in ß cells, possibly by sharing common pathways of ß cell dysfunction. TAC-induced ß cell dysfunction is potentially reversible. Inhibition of the calcineurin-NFAT pathway may contribute to the diabetogenic effect of CNIs but does not explain the stronger effect of TAC compared with CsA.

High frequency de novo alterations in the long-range genomic structure of the mouse pseudoautosomal region.

The pseudoautosomal region (PAR) is a segment of shared homology between the X and Y chromosomes. Here we report physical linkage of three mouse PAR probes: DXYHgu1, DXYMov15 and (TTAGGG)n. Steroid sulphatase (Sts) maps distal to these probes, indicating that there is an internal array of the telomere sequence (TTAGGG)n in the PAR. Pseudoautosomal PacI restriction fragments, up to 2 Mb in size, are unstable in C57BL/6 x C57BL/6 crosses. New alleles, often several hundred kilobases different in size, occur at a sex-averaged rate of approximately 30% per allele. Such frequent large-scale germline genome arrangements are without precedent in mammals.

Expression of the X-inactivation-associated gene XIST during spermatogenesis.

Mammalian X-chromosome inactivation is thought to be controlled by the X inactivation centre (XIC, X-controlling element -Xce-in mice). A human gene, XIST and its mouse counterpart, Xist, which map to the XIC/Xce, are expressed exclusively from inactive X chromosomes, suggesting their involvement in the process of X-inactivation. We now report the presence of Xist/XIST transcripts in newborn and adult mouse testes, and in human testicular tissue with normal spermatogenesis, but not in the testes of patients who lack germ cells. Our results indicate that while the X chromosome in males is active in somatic cells, it undergoes inactivation during spermatogenesis.

Cloning and expression of the mouse pseudoautosomal steroid sulphatase gene (Sts).

Steroid sulphatase (STS) is an important enzyme in steroid metabolism. The human STS gene has been cloned and mapped to Xp22.3, proximal to the pseudoautosomal region (PAR). Using quantitative differences in STS activity among various mouse strains, a segregation pattern consistent with autosomal linkage was first reported, but more recent studies have linked Sts to the mouse PAR. Failed attempts to clone the mouse Sts gene using human reagants (STS cDNA and anti-STS antibodies) suggest a substantial divergence between these genes. However, partial amino-terminal sequence from purified rat liver Sts is very similar to its human counterpart, and several domains are conserved among all the sulphatases. We followed a degenerate-primer reverse transcriptase-PCR (RT-PCR) approach to amplify a conserved fragment of the rat Sts cDNA that was then used to clone the mouse Sts cDNA. This 2.3-kb cDNA revealed 75% similarity with rat Sts cDNA, while it was only 63% similar to human STS cDNA. Transfection of STS(-) A9 cells with the mouse Sts cDNA restored STS enzymatic activity. Sts was also mapped physically to the distal end of the mouse sex chromosomes, and our backcross studies placed Sts distal to the 'obligatory' cross-over in male meiosis.

The murine Xe169 gene escapes X-inactivation like its human homologue.

Among a number of genes that escape X-chromosome inactivation in humans, three have been evaluated in mice and unexpectedly all three are subject to X-inactivation. We report here the cloning and expression studies of a novel mouse gene, Xe169, and show that it escapes X-inactivation like its human homologue. Xe169 was assigned to band F2/F3 on the mouse X chromosome by fluorescent in situ hybridization and Southern analysis indicates that the gene is located outside the pseudoautosomal region. Homologous, but divergent, sequences exist on the Y chromosome. In vitro and in vivo studies show that Xe169 is expressed from both the active and the inactive X chromosomes. Xe169 is the first cloned non-pseudoautosomal gene that escapes X-inactivation in mice.

Epidemiology of systemic lupus erythematosus in Asia.

In this paper we aim to review the prevalence, incidence and disease presentation of lupus patients in Asian populations. The database of the National Library of Medicine (MEDLINE) through PUBMED up to September 2009 was searched for relevant articles using the search terms 'systemic lupus erythematosus', 'epidemiology', and 'Asia'. Articles on lupus prevalence, incidence, and clinical manifestations were retrieved and their bibliographies further screened for relevant articles. Proceedings of national, regional, and international rheumatology and SLE conferences were likewise searched for relevant abstracts. Retrieval rate of relevant articles was approximately 80%. Prevalence and incidence figures are expressed per 100,000 populations. Among the articles reviewed, epidemiologic information was usually obtained through population surveys or hospital cohorts. Prevalence data are available from 24 countries, and generally fall within 30-50/100,000 population. However, one survey showed a higher prevalence of 70 (Shanghai), while three others showed a lower prevalence of 3.2-19.3 (India, Japan, Saudi Arabia). Only three countries have recorded incidence rates and these varied from 0.9 to 3.1 per annum. Thirty articles from 21 countries describe lupus disease presentation. Common manifestations include mucocutaneous lesions (seen in 52-98% of patients) and arthritis/musculoskeletal complaints (36-95%). Antinuclear antibodies were generally positive in 89-100% of patients, except for two studies. Renal involvement ranged from 18% to 100% with most articles reporting this in >50% of their patients. Discoid lesions, serositis, and neurologic involvement were the least frequently seen symptoms. There is varying epidemiologic information regarding systemic lupus erythematosus among countries in Asia. Prevalence rates usually fall within 30-50/100,000 population. Incidence rates, as reported from three countries, varied from 0.9/100,000 to 3.1% per annum. It is difficult to make generalizations about how the epidemiologic character of the disease varies from country to country. However, similarities in disease manifestations can be observed. .

Correction: DUB3 and USP7 de-ubiquitinating enzymes control replication inhibitor Geminin: molecular characterization and associations with breast cancer.

The final sentence of the Acknowledgements should be as follows: This work was supported by grants from Instituto de Salud Carlos III (BA15/00092), Spanish Ministry of Economy and Competitiveness/EU-ERDF (SAF2016-80626-R, SAF2013-49149-R, BFU2014-51672-REDC), Fundación CajaCanarias (AP2015/008) to RF, and the Australian National Health and Medical Research (NHMRC program grant to SRL and KKK (APP1017028).

Low-dose-rate brachytherapy for prostate cancer in renal transplant recipients.

PURPOSE: Prostate cancer (PCa) is the most common malignancy among men and one of the most common neoplasms affecting renal transplant recipients (RTRs). The available treatments for localized PCa among the general population (GP), surgery and external beam radiotherapy, carry a risk of damage to the transplanted kidney, the ureters, and the bladder and therefore tend to be avoided by most groups. The objective of this study was to assess the efficacy and feasibility of low-dose-rate brachytherapy (LDR-BT) for PCa in RTRs. METHODS AND MATERIALS: We carried out a retrospective review on all RTRs diagnosed of PCa who had undergone LDR-BT at our institution between 2000 and 2015. Nine patients met these criteria, but 1 did not fulfill the followup. Hence, we analyzed 8 patients. We reviewed all clinical data for PCa and graft function in these patients and compared the results with the GP. RESULTS: Mean baseline prostate-specific antigen was 6.8 ± 1.9 ng/mL. All PCa had a Gleason score of 6 and were classified as low risk according the Europe Association of Urology guidelines. Mean followup after seed implantation was 48 ± 12.8 months. All 8 patients remain free of prostate-specific antigen failure. Five-year progression-free survival, cancer-specific survival, and overall survival rates were 100%, 100%, and 62.5%. There was no specific toxicity associated with LDR-BT, and there were no acute adverse events affecting the graft. CONCLUSIONS: LDR-BT is a feasible and acceptable treatment for localized PCa in RTRs. Oncological outcomes are similar to the GP, and there is minimal toxicity to the renal graft.

DUB3 and USP7 de-ubiquitinating enzymes control replication inhibitor Geminin: molecular characterization and associations with breast cancer.

Correct control of DNA replication is crucial to maintain genomic stability in dividing cells. Inappropriate re-licensing of replicated origins is associated with chromosomal instability (CIN), a hallmark of cancer progression that at the same time provides potential opportunities for therapeutic intervention. Geminin is a critical inhibitor of the DNA replication licensing factor Cdt1. To properly achieve its functions, Geminin levels are tightly regulated through the cell cycle by ubiquitin-dependent proteasomal degradation, but the de-ubiquitinating enzymes (DUBs) involved had not been identified. Here we report that DUB3 and USP7 control human Geminin. Overexpression of either DUB3 or USP7 increases Geminin levels through reduced ubiquitination. Conversely, depletion of DUB3 or USP7 reduces Geminin levels, and DUB3 knockdown increases re-replication events, analogous to the effect of Geminin depletion. In exploring potential clinical implications, we found that USP7 and Geminin are strongly correlated in a cohort of invasive breast cancers (P<1.01E-08). As expected, Geminin expression is highly prognostic. Interestingly, we found a non-monotonic relationship between USP7 and breast cancer-specific survival, with both very low or high levels of USP7 associated with poor outcome, independent of estrogen receptor status. Altogether, our data identify DUB3 and USP7 as factors that regulate DNA replication by controlling Geminin protein stability, and suggest that USP7 may be involved in Geminin dysregulation during breast cancer progression.

DUB3 and USP7 de-ubiquitinating enzymes control replication inhibitor Geminin: molecular characterization and associations with breast cancer.

This corrects the article DOI: 10.1038/onc.2017.21.

Changes in the epidemiology of meticillin-resistant Staphylococcus aureus associated with the emergence of EMRSA-16 at a university hospital.

This study investigated the molecular epidemiology of meticillin-resistant Staphylococcus aureus (MRSA) in the University Hospital of the Canary Islands (HUC) in order to evaluate epidemiological changes over a six-year period. Clinical and epidemiological data were collected between May 2000 and December 2003, and isolates were subjected to pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), SCCmec typing and spa typing. Since 2000, the rate of MRSA infections has increased at the HUC, coinciding with the emergence and spread of the EMRSA-16 clone (ST36-MRSA-II) and replacement of the Iberian clone (ST247-MRSA-I). Genotypic changes were associated with changes in the epidemiological profile. The mean age and proportion of patients over 60 years old (P=0.01) and the proportion of respiratory infections (P=0.001) increased significantly. Gentamicin and tetracycline susceptibility of MRSA isolates increased (P<0.001) following the emergence of EMRSA-16. Combining PFGE, SCCmec and MLST has been instrumental in understanding these changes and defining the clones circulating in the HUC patient population.

[Renal oncocytoma. Review of our 22 patients]. / Oncocitoma renal. Revisión de nuestra serie de 22 pacientes.

UNLABELLED: Renal oncocytoma (OR) is a benign tumor. It may represent up to 3-7% of solid kidney masses, and shows specifics cellular and evolutive characteristics. Metacronicity, multifocality and bilateralism has been reported. MATERIALS AND METHODS: Between 1986 and 2005, 478 kidney tumors have been surgically treated at our institution. We report the frequency and characteristics of OR in our patients, compared with renal cell carcinomas (RCC). We try to find out the rate of multifocality, bilateralism and other tumor association, and the number of neoplasms originally diagnosed as OR before surgery. Mean and median follow up: 36.86 and 13 months (1-193). Specific survival rate 100%. RESULTS: We found 24 OR in 10 men and 12 women with a mean age of 59 years (34-84). 12 in the left kidney and 12 in the right one, one patient presenting oncocytomatosis. Tumor mean size was 4.64 cm (1-12.5 cm). Tumors were discovered incidentally in 17 cases. Presentation symptoms in the rest of patients were gross hematuria. CONCLUSIONS: The rate of OR found in our sample population of renal tumors undergoing surgery matches other series already published. Two synchronic OR, but not metacronous, bilateral or metastatic tumors were found. All cases presented a benign evolution.

Mouse p73 gene maps to the distal part of chromosome 4 and might be involved in the progression of gamma-radiation-induced T-cell lymphomas.

We have isolated and sequenced a DNA fragment of about 12 kb that comprises exons 5-14 of the mouse p73 gene. We have identified four polymorphic markers, and one of them has been used to genetically map p73 to the distal part of chromosome 4. Previously, we have reported that gamma-radiation-induced T-cell lymphomas undergo frequent loss of heterozygosity around marker D4Mit205b at the distal part of chromosome 4. Based on this, we have performed loss of heterozygosity analysis in a set of T-cell lymphomas, and we have found allelic losses of p73 in 32.6% (16 of 49) of the tumors analyzed. Interestingly, allelic losses occur concurrently at both p73 and D4Mit205b, thus suggesting that p73 could be specifically inactivated in radiation-induced T-cell lymphomas.

[Complex vesicoureteral reflux. Our experience]. / Reflujo vesicoureteral complejo. Revisión de nuestra serie.

OBJECTIVE: To analize the proportion of complex reflux in the whole amount of patients treated endoscopically of vesicoureteral reflux in our hospital. To determine the endoscopic treatment success in complex reflux, and the influence of reflux grade in it. MATERIAL AND METHODS: We present our experience between 1992 and 2003 with three kinds of substances (polytetrafluoroethylene, polydimethylsiloxane and dextranomer-hyaluronic acid copolymer). We treated complex reflux in 74 patients with endoscopic injection. All patients were scheduled to have voiding cystourethrogram 3 and 9 moths after injection. A positive response was defined as grade 0 or I reflux. RESULTS: Reflux was solved using the endoscopic procedure in 86.25% after first injection, 93.75% after second and 96.25% after third. The corresponding results for reflux grade II, III and IV were 88.9%, 83.3% and 100%. CONCLUSIONS: We conclude that subureteral injection of different sustances (Teflon, Macroplastique or Deflux) is a useful treatment for most cases of vesicoureteral reflux. We propose it as first step of treatment.

[Cystic ureteritis in a kidney transplantation candidate]. / Ureteritis quística en paciente candidata a trasplante renal.

Cystic ureteritis is a very uncommon pathology, whose pathogenesis is not well established. It is usually asociated with chronic infectious factors. It presents unspecific symptoms but characteristic radiologic findings. There is not an especific treatment for this disease. Kidney transplant is the final pathway for patients with chronic renal failure. We report a case of cystic ureteritis diagnosed during pre-transplant study.

[Xanthogranulomatous pyelonephritis in childhood. A case report]. / Pielonefritis xantogranulomatosa en la infancia. A propósito de un caso.

Xanthogranulomatous pyelonephritis is an uncommon chronic inflammatory renal disorder. Most cases have been described in middle aged women and it is extremely rare in children. We report a case of a 4 year old girl who suffered from recurrent urinary tract infection and antibiotic therapy resistance. She had low growth-rate and palpable left renal mass on examination and was confirmed by radiological findings. We performed left kidney partial resection and then histological examination showed focal xanthogranulomatous pyelonephritis. The focal form of the disease may respond to antibiotic treatment although usually an enucleation or partial resection must be performed. In conclusion xanthogranulomatous pyelonephritis should be considered in the differential diagnosis of a renal mass and recurrent urinary tract infection in childhood.

Expression of steroid sulfatase during embryogenesis.

Neurosteroids are steroids that are synthesized de novo in the brain from cholesterol and, in general, mediate their effects through ion-gated channel receptors such as gamma-aminobutyric acidA (GABA[A]) and N-methyl-D-aspartate receptors rather than through classical nuclear steroid hormone receptors. Steroid hormones are known to exist not only as free compounds, but also as sulfated derivatives. Pharmacological studies indicate that unconjugated and sulfated steroids, such as pregnenolone and pregnenolone sulfate, may have opposite effects on GABA(A) receptors. Thus, pregnenolone acts as a potent positive allosteric modulator of gamma-aminobutyric acid action at GABA(A )receptors, whereas pregnenolone sulfate acts as a potent negative modulator. Recent experiments also suggest that dehydroepiandrosterone and dehydroepiandrosterone sulfate may have distinct effects on growth of neurites from embryonic neocortical neurons in vitro. Thus, regulation of steroid sulfation may have profound behavioral and morphological effects on the nervous system. We, therefore, studied the developmental expression of the enzyme steroid sulfatase (STS), which converts sulfated steroids to free steroids. By in situ hybridization, STS messenger RNA was expressed in the embryonic mouse cortex, hindbrain, and thalamus during the last third of gestation. The sites of expression of STS were similar to those of P450c17, suggesting that these two enzymes may have concerted actions in similar functional processes.

Effect of thyroxine administration on the expression of epidermal growth factor in the kidney and submandibular gland of neonatal mice. An immunocytochemical and in situ hybridization study.

The effect of T4 administration on epidermal growth factor (EGF) expression in kidney and submandibular glands (SMG) was studied in newborn mice. EGF messenger RNA (mRNA) abundance and EGF immunoreactivity were assessed by in situ hybridization and immunohistochemical techniques, respectively. T4 treatment on days 0-6 augmented both the in situ hybridization and immunocytochemical signals in kidney but not in SMG on day 7. By contrast, T4 injections on days 7-14 did not alter the in situ hybridization or immunocytochemical signals in kidney but increased both signals in SMG on day 15. Thus, neonatal T4 treatment augments the levels of EGF mRNA as well as EGF immunoreactivity in both kidney and SMG. The T4 effect is manifested during the first week in kidney whereas the SMG responds to T4 treatment only during the second week of life.

Steroid sulfatase expression in human placenta: immunocytochemistry and in situ hybridization study.

Steroid sulfatase (STS), an important enzyme in the pathway of estrogen synthesis from sulfated steroid precursors, was localized to the syncytial trophoblast of human placentas during different periods of pregnancy by using a mouse monoclonal antibody and immunocytochemical techniques. Preembedding immunoelectron microscopy revealed STS immunoreactivity associated with the rough endoplasmic reticulum of the syncytial trophoblast. STS mRNA was also localized to this outermost layer of the human trophoblast. At a cellular level, both STS message and immunoreactivity seemed to be more abundant during the late first and early second trimesters than in term placentas. Although the syncytial trophoblast is known to originate from the cytotrophoblast, neither STS immunoreactivity nor STS mRNA was detected in the cytotrophoblast at any stage of placental development studied.

Characterization of the mouse Dazap1 gene encoding an RNA-binding protein that interacts with infertility factors DAZ and DAZL.

BACKGROUND: DAZAP1 (DAZ Associated Protein 1) was originally identified by a yeast two-hybrid system through its interaction with a putative male infertility factor, DAZ (Deleted in Azoospermia). In vitro, DAZAP1 interacts with both the Y chromosome-encoded DAZ and an autosome-encoded DAZ-like protein, DAZL. DAZAP1 contains two RNA-binding domains (RBDs) and a proline-rich C-terminal portion, and is expressed most abundantly in the testis. To understand the biological function of DAZAP1 and the significance of its interaction with DAZ and DAZL, we isolated and characterized the mouse Dazap1 gene, and studied its expression and the subcellular localization of its protein product. RESULTS: The human and mouse genes have similar genomic structures and map to syntenic chromosomal regions. The mouse and human DAZAP1 proteins share 98% identity and their sequences are highly similar to the Xenopus orthologue Prrp, especially in the RBDs. Dazap1 is expressed throughout testis development. Western blot detects a single 45 kD DAZAP1 protein that is most abundant in the testis. Although a majority of DAZAP1 is present in the cytoplasmic fraction, they are not associated with polyribosomes. CONCLUSIONS: DAZAP1 is evolutionarily highly conserved. Its predominant expression in testes suggests a role in spermatogenesis. Its subcellular localization indicates that it is not directly involved in mRNA translation.