ST18 affects cell-cell adhesion in pemphigus vulgaris in a tumour necrosis factor-α-dependent fashion.

BACKGROUND: Pemphigus vulgaris (PV) is a life-threatening mucocutaneous autoimmune blistering disease. We previously showed that genetic variants within the ST18 gene promoter area confer a sixfold increase in the propensity to develop PV. ST18, a transcription factor, was found to be overexpressed in the epidermis of patients with PV. In addition, it was found to promote autoantibody-mediated abnormal epidermal cell-cell adhesion and secretion of proinflammatory mediators by keratinocytes. OBJECTIVES: To delineate the mechanism through which ST18 contributes to destabilization of cell-cell adhesion. METHODS: We used quantitative reverse-transcriptase polymerase chain reaction, immunofluorescence microscopy, a luciferase reporter system, site-directed mutagenesis, chromatin immunoprecipitation (ChIP) and the dispase dissociation assay. RESULTS: The ChIP and luciferase reporter assays showed that ST18 directly binds and activates the TNF promoter. Accordingly, increased ST18 expression contributes to PV pathogenesis by destabilizing cell-cell adhesion in a tumour necrosis factor (TNF)-α-dependent fashion. In addition, dual immunofluorescence staining showed increased expression of both ST18 and TNF-α in the skin of patients with PV carrying an ST18-associated PV risk variant, which was found to be associated with a more extensive PV phenotype. CONCLUSIONS: Our findings suggest a role for TNF-α in mediating the deleterious effect of increased ST18 expression in PV skin.

Molecular epidemiology of pachyonychia congenita in the Israeli population.

BACKGROUND: Pachyonychia congenita (PC) is a rare autosomal dominant disorder featuring palmoplantar keratoderma, nail dystrophy, oral leucokeratosis, pilosebaceous cysts and natal teeth. PC results from dominant mutations in one of five genes (KRT6A, KRT6B, KRT6C, KRT16, KRT17) encoding keratin proteins. AIM: To delineate the clinical and genetic features of PC in a series of Israeli patients. METHODS: We used direct sequencing of genomic DNA, and also used cDNA sequencing where applicable. RESULTS: We collected clinical information and molecular data in a cohort of Israeli families diagnosed with PC (n = 16). Most of the patients were Ashkenazi Jews and had a family history of PC. The most common clinical findings were painful focal plantar keratoderma (94%) accompanied by nail dystrophy (81%), pilosebaceous cysts (31%) and prenatal/natal teeth (13%). In contrast to the high prevalence of KRT6A mutations in other populations, we found that KRT16 mutations were the most common type among Israeli patients with PC (56%). Most (77%) of the Israeli patients with PC with KRT16 mutation carried the same variant (c.380G>A; p.R127H) and shared the same haplotype around the KRT16 locus, suggestive of a founder effect. CONCLUSION: The data gleaned from this study emphasizes the importance of population-specific tailored diagnostic strategies.

Phylogenetic classification and clinical aspects of a new putative Deltapapillomavirus associated with skin lesions in cattle.

Bovine papillomaviruses (BPVs) are recognized as causal agents of benign and malignant tumors in cattle. Thirteen types of BPVs have already been described and classified into 3 distinct genera. Divergences in the nucleotide sequence of the L1 gene are used to identify new viral types through the employment of PCR assays with degenerated primers. In the present study, a method for identifying BPVs based on PCR-RFLP and DNA sequencing allowed the identification of a new putative Deltapapillomavirus, designated JN/3SP (JQ280500.1). The analysis of the L1 gene showed that this strain was most closely related to the BPVs -1, -2, -13 , and OaPV1 (71-73% genetic similarity). In this study, we describe the detection of this new putative Deltapapillomavirus type and verify its phylogenetic position within the genus.

Chromosome aberrations in cells infected with bovine papillomavirus: comparing cutaneous papilloma, esophagus papilloma, and urinary bladder lesion cells.

THE MAJORITY OF MALIGNANT CELLS PRESENT GENETIC INSTABILITY WITH CHROMOSOME NUMBER CHANGES PLUS SEGMENTAL DEFECTS: these changes involve intact chromosomes and breakage-induced alterations. Some pathways of chromosomal instability have been proposed as random breakage, telomere fusion, and centromere fission. Chromosome alterations in tumor cells have been described in animal models and in vitro experiments. One important question is about possible discrepancies between animal models, in vitro studies, and the real events in cancer cells in vivo. Papillomaviruses are relevant agents in oncogenic processes related to action on host genome. Recently, many reports have discussed the presence of virus DNA in peripheral blood, in humans and in animals infected by papillomaviruses. The meaning of this event is of controversy: possible product of apoptosis occurring in cancer cells, metastasized cancer cells, or active DNA sequences circulating in bloodstream. This study compares chromosome aberrations detected in bovine cells, in peripheral blood cells, and in BPV lesion cells: the literature is poor in this type of study. Comparing chromosome aberrations described in the different cells, a common mechanism in their origin, can be suggested. Furthermore blood cells can be evaluated as an effective way of virus transmission.

Bovine papillomavirus in Brazil: detection of coinfection of unusual types by a PCR-RFLP method.

Bovine papillomavirus (BPV) is recognized as a causal agent of benign and malignant tumors in cattle. Thirteen types of BPV are currently characterized and classified into three distinct genera, associated with different pathological outcomes. The described BPV types as well as other putative ones have been demonstrated by molecular biology methods, mainly by the employment of degenerated PCR primers. Specifically, divergences in the nucleotide sequence of the L1 gene are useful for the identification and classification of new papillomavirus types. On the present work, a method based on the PCR-RFLP technique and DNA sequencing was evaluated as a screening tool, allowing for the detection of two relatively rare types of BPV in lesions samples from a six-year-old Holstein dairy cow, chronically affected with cutaneous papillomatosis. These findings point to the dissemination of BPVs with unclear pathogenic potential, since two relatively rare, new described BPV types, which were first characterized in Japan, were also detected in Brazil.

Fetal middle cerebral artery Doppler fluctuations after laser surgery for twin-twin transfusion syndrome.

OBJECTIVE: The objective of this study was to compare alterations in the middle cerebral artery (MCA) pulsatility index (PI) and mean velocity (V (mean)) after laser surgery for twin-twin transfusion syndrome (TTTS). STUDY DESIGN: MCA Doppler studies were conducted 1 day before and after laser surgery for TTTS. The pre- and postoperative mean (standard deviation) of the MCA PI and V (mean) z-scores of the recipient and donor fetuses were calculated and compared. Data were analyzed using paired testing and multivariable linear regression models. RESULTS: A total of 103 patients met the study criteria. Recipients' MCA PI increased from -1.29 (1.20) preoperatively to 0.14 (1.52) postoperatively (P < 0.0001), whereas the donors' PI did not change significantly (-0.31 (1.67) to -0.67 (1.29); P = 0.12). There was no significant difference between preoperative and postoperative MCA V (mean) in donors (0.39 (0.83) and 0.38 (0.93), respectively; P = 0.5048) or recipients (0.60 (0.74) and 0.63 (0.90), respectively; P = 0.5324). CONCLUSIONS: Despite the changes in the MCA PI after laser surgery for TTTS, the MCA V (mean) remained constant. These findings may suggest some autoregulatory capacity in the cerebral vessels of the mid-trimester fetus.

Residual vascular communications in twin-twin transfusion syndrome treated with sequential laser surgery: frequency and clinical implications.

OBJECTIVES: The goal of fetoscopic laser surgery for twin-twin transfusion syndrome (TTTS) is to ablate all placental vascular communications, thereby separating the fetal circulatory systems. We sought to ascertain the frequency and clinical implications of residual vascular communications (RVC) post preferential sequential selective laser photocoagulation of communicating vessels (SQLPCV). STUDY DESIGN: TTTS placentas treated via preferential SQLPCV were examined. Patency of vascular communications was assessed via water and/or milk injections. Cases with intrauterine fetal demise or placental disruption were excluded. Outcomes with and without RVC were compared. RESULTS: One hundred seventy-four TTTS patients were treated during the study period. Dual survival at birth was 76% (133/174). Of the 133 dual survivors, 105 (79%) submitted an intact placenta. Five of these 105 placentas had RVC (4.8%). Comparison of RVC versus non-RVC cases revealed the following: gestational age at delivery 28.7(6.5) vs. 33.4(3.3) weeks (p=0.178); recipient birth weight 1287(1061) vs. 1973(610) grams (p=0.020); donor birth weight 1429(1369) vs. 1653(715) grams (p=0.518); donor central/eccentric placental cord insertion 80% vs. 17% (p=0.006). One case required a second laser surgery to complete the laser ablation; this placenta did not have RVC after delivery. Otherwise there were no cases of persistent TTTS. One of the 5 RVC cases (20%) exhibited neonatal findings consistent with twin anemia-polycythemia sequence (TAPS), while none of the non-RVC cases had TAPS (p=0.005). CONCLUSIONS: The rate of RVC was less than 5% among gestations with dual survivors post preferential SQLPCV treatment for TTTS.

Fetal heart rate changes associated with sequential selective laser surgery for twin-twin transfusion syndrome.

OBJECTIVE: To investigate perioperative changes in fetal heart rate (FHR) associated with sequential vs standard selective laser photocoagulation of communicating vessels for the treatment of twin-twin transfusion syndrome (TTTS). STUDY DESIGN: Women with TTTS were treated with the intent of using the sequential procedure. Those who failed this treatment were categorized as having undergone the standard procedure. Pre- and postoperative FHR of donor and recipient fetuses were analyzed. RESULT: Of 98 women, 35 received the standard technique. A postoperative drop in the mean donor FHR was observed in gestations receiving the standard laser, but not in those receiving the sequential technique. In multivariable models that included operative and gestational characteristics, the use of the sequential treatment was associated with improved stability of the FHR of the donor twin. CONCLUSION: The stability in donor FHR following sequential laser ablation when compared with the standard technique is consistent with improved donor hemodynamics.

Liver X receptor alpha regulates fatty acid synthase expression in chicken.

Liver X receptor alpha (LXRalpha), also referred to as nuclear receptor subfamily 1, group H, member 3 is a member of the nuclear hormone receptor superfamily, and has recently been shown to act as a master transcription factor governing hepatic lipogenesis in mammals. Liver X receptor alpha directly regulates both the expression of other lipogenic transcription factors and the expression of lipogenic enzymes, thereby enhancing hepatic fatty acid synthesis (FASN). In birds, like in humans, fatty acid synthesis primarily occurs in the liver. Whether LXRalpha is involved in hepatic regulation of lipogenic genes remained to be investigated in this species. Here we show that fatty acid synthase and the expression of other lipogenic genes (sterol regulatory element binding protein 1 and steroyl coenzyme A desaturase 1) are induced in chicken hepatoma cells in response to a pharmacological liver X receptor agonist, T0901317. A detailed analysis of the chicken FASN promoter revealed a functional liver X response element. These data define the chicken FASN gene as a direct target of LXRalpha and further expand the role of LXRalpha as a regulator of lipid metabolism in this species.

New insights into the structural characteristics of the arabinogalactan-protein (AGP) fraction of gum arabic.

The structural characteristics of the gum exudate of Acacia senegal (gum arabic) have been investigated by monitoring the composition and physicochemical properties before and after treatment with proteolytic enzyme and various alkaline systems. Molecular mass ( M w) and radius of gyration ( R g) measurements were performed using gel permeation chromatography (GPC) coupled to refractive index, UV absorbance, and multiangle light scattering detectors and indicated that the macromolecules present have a compact structure. It was found that treatment with proteolytic enzyme caused the arabinogalactan-protein component (AGP) with average molecular mass approximately 2 x 10 (6) Da to degrade, yielding material of molecular mass approximately 4 x 10 (5) Da, whereas the bulk of the material corresponding to the protein-deficient arabinogalactan component (AG) with molecular mass 4 x 10 (5) remained unaffected. Barium hydroxide was found to hydrolyze the polysaccharide component (AG) itself in addition to the proteinaceous component as demonstrated in control experiments using dextran. However, sodium borohydride/sodium hydroxide treatments were unable to hydrolyze dextran and were assumed to hydrolyze only the proteinaceous component of gum arabic. The AGP component was completely degraded, yielding material of molecular mass approximately 4.5 x 10 (4) Da. It has been concluded, therefore, that the enzyme did not fully hydrolyze all of the protein present and that the AGP component of gum arabic consists of carbohydrate blocks of approximately 4.5 x 10 (4) Da linked to a polypeptide chain consistent with the wattle blossom structure. Because the AGP was degraded to differing extents using a mild and more severe sodium borohydride/sodium hydroxide treatment, it was concluded that the polysaccharide moieties were linked through both O-serine and O-hydroxyproline residues. The gum arabic sample was deglycosylated by treatment with anhydrous hydrogen fluoride and revealed the presence of two putative core proteins of approximately 3 x 10 (4) and approximately 5 x 10 (3) Da, respectively, which correspond to proteins of approximately 250 and 45 amino acids in length. A new model for the structure of the AGP component has been proposed.

Endotoxemic-like shock induced by Loxosceles spider venoms: pathological changes and putative cytokine mediators.

The systemic symptoms, tissue lesions and release of cytokines were analysed in four isogenic mouse strains with distinct haplotypes injected with various doses of Loxosceles intermedia spider venom. The estimated LD50 were 24.5 microg for C57Bl/6, 17.6 microg for BALB/c, 6.3 microg for C3H/HeJ and 4.6 microg for A/Sn mice. Prostration, acute cachexia, hypothermia, neurological disorders and hemoglobinuria were the signals preceding death. Accumulation of eosinophilic material inside the proximal and distal renal tubules and acute tubular necrosis were the most common histopathological findings. Death was prevented by previous treatment of venom with specific antivenom serum. The protein F35 purified from the whole venom retained the ability to induce the symptoms of the whole venom. The cytokines tumor necrosis factor (TNF), interleukins IL-6 and IL-10 and the radical nitric oxide were detected in serum at different levels after venom injection. These findings indicate that the state of shock produced in mice by whole endotoxin-free L. intermedia venom or by its purified fraction, protein F35, mimics the endotoxemic shock, that susceptibility to the systemic effects of the venom varies among mice of different haplotypes and that the pattern of in vivo cytokine release resembles that of endotoxemic shock.