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Background: Since it is currently not possible to eradicate hepatitis B virus (HBV) infection with existing treatments, research continues to uncover new therapeutic strategies. HBV core protein, encoded by the HBV core gene ( HBC ), intervenes in both structural and functional processes, and is a key protein in the HBV life cycle. For this reason, both the protein and the gene could be valuable targets for new therapeutic and diagnostic strategies. Moreover, alterations in the protein sequence could serve as potential markers of disease progression., Aim: To detect, by next-generation sequencing, HBC hyper-conserved regions that could potentially be prognostic factors and targets for new therapies., Methods: Thirty-eight of 45 patients with chronic HBV initially selected were included and grouped according to liver disease stage [chronic hepatitis B infection without liver damage (CHB, n = 16), liver cirrhosis (LC, n = 5), and hepatocellular carcinoma (HCC, n = 17)]. HBV DNA was extracted from patients' plasma. A region between nucleotide (nt) 1863 and 2483, which includes HBC , was amplified and analyzed by next-generation sequencing (Illumina MiSeq platform). Sequences were genotyped by distance-based discriminant analysis. General and intergroup nt and amino acid (aa) conservation was determined by sliding window analysis. The presence of nt insertion and deletions and/or aa substitutions in the different groups was determined by aligning the sequences with genotype-specific consensus sequences., Results: Three nt (nt 1900-1929, 2249-2284, 2364-2398) and 2 aa (aa 117-120, 159-167) hyper-conserved regions were shared by all the clinical groups. All groups showed a similar pattern of conservation, except for five nt regions (nt 1946-1992, 2060-2095, 2145-2175, 2230-2250, 2270-2293) and one aa region (aa 140-160), where CHB and LC, respectively, were less conserved ( P < 0.05). Some group-specific conserved regions were also observed at both nt (2306-2334 in CHB and 1935-1976 and 2402-2435 in LC) and aa (between aa 98-103 in CHB and 28-30 and 51-54 in LC) levels. No differences in insertion and deletions frequencies were observed. An aa substitution (P79Q) was observed in the HCC group with a median (interquartile range) frequency of 15.82 (0-78.88) vs 0 (0-0) in the other groups ( P < 0.05 vs CHB group)., Conclusion: The differentially conserved HBC and HBV core protein regions and the P79Q substitution could be involved in disease progression. The hyper-conserved regions detected could be targets for future therapeutic and diagnostic strategies., Competing Interests: Conflict-of-interest statement: Josep Gregori is an employee of Roche Diagnostics, SL., (©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.)

Aim: To detect hyper-conserved regions in the hepatitis B virus (HBV) X gene ( HBX ) 5' region that could be candidates for gene therapy., Methods: The study included 27 chronic hepatitis B treatment-naive patients in various clinical stages (from chronic infection to cirrhosis and hepatocellular carcinoma, both HBeAg-negative and HBeAg-positive), and infected with HBV genotypes A-F and H. In a serum sample from each patient with viremia > 3.5 log IU/mL, the HBX 5' end region [nucleotide (nt) 1255-1611] was PCR-amplified and submitted to next-generation sequencing (NGS). We assessed genotype variants by phylogenetic analysis, and evaluated conservation of this region by calculating the information content of each nucleotide position in a multiple alignment of all unique sequences (haplotypes) obtained by NGS. Conservation at the HBx protein amino acid (aa) level was also analyzed., Results: NGS yielded 1333069 sequences from the 27 samples, with a median of 4578 sequences/sample (2487-9279, IQR 2817). In 14/27 patients (51.8%), phylogenetic analysis of viral nucleotide haplotypes showed a complex mixture of genotypic variants. Analysis of the information content in the haplotype multiple alignments detected 2 hyper-conserved nucleotide regions, one in the HBX upstream non-coding region (nt 1255-1286) and the other in the 5' end coding region (nt 1519-1603). This last region coded for a conserved amino acid region (aa 63-76) that partially overlaps a Kunitz-like domain., Conclusion: Two hyper-conserved regions detected in the HBX 5' end may be of value for targeted gene therapy, regardless of the patients' clinical stage or HBV genotype., Competing Interests: Conflict-of-interest statement: Josep Gregori is an employee of Roche Diagnostics, SL.

Objective: Metallic stents, sometimes augmented by "J" endostents, were placed in ureters obstructed by advanced carcinoma of the cervix to improve or maintain renal function in these patients at a level necessary to pursue long-term chemotherapy and radiation therapy., Subjects and Methods: Seventeen ureters in 11 patients were treated with metallic stents, which were placed by an anterograde approach in 10 patients and by a retrograde approach in one. An anterograde ureteroneocystostomy was necessary in two of the 11 patients. J endostents were placed for I month in all patients. In 10 ureters, J endostents were reintroduced during follow-up 1-48 months later., Results: In 17 ureters, metallic stents were successfully placed; in the remaining three ureters, the procedures were aborted because of technical difficulties. The goal of improving and maintaining renal function was met in 10 patients. Serum creatinine levels returned to normal in four patients and stabilized at a range of 2.7-3.2 mg/dl in six more patients, permitting treatment with chemotherapy, radiation therapy, or both. Four of the 11 patients died within 4 months after treatment, and three more died within 2 years. At the time of death or last follow-up, seven metallic stents and eight combinations of metallic stents and J endostents were patent. Complications were minor., Conclusion: Placement of metallic stents is recommended as an effective method to ensure patency and to facilitate cystoscopic replacement of J endostents in ureters compromised by carcinoma of the cervix. Renal function can be maintained, allowing chemotherapy. Survival is governed by the effectiveness of chemotherapy and radiation therapy.

As actinomycetoma is more frequent in males than in females, the possibility that hormones might modify the Nocardia brasiliensis growth and the course of experimental actinomycetoma was explored. Five N. brasiliensis strains were grown on Sabouraud agar containing estradiol, progesterone or testosterone, in 3 different concentrations. Colony diameters were measured weekly for 7 weeks. N. brasiliensis strains were also grown in Sabouraud broth containing hormones. Glucose concentration was measured weekly for 6 weeks. Finally, experimental actinomycetoma was produced in male and female hormone-treated mice. Invasion rate, plantar pad diameter and positive retrocultures were assessed. In vitro experiments showed that progesterone and testosterone inhibit N. brasiliensis growth, manifested by lower colony diameters and greater glucose concentrations. In vivo experiments demonstrated that estradiol limits actinomycetoma development. Progesterone and testosterone induced greater diameters of inoculated plantar pads and greater invasion rates with greater positive culture numbers than estradiol. Results partially explain the resistance of females to actinomycetoma.

To determine the effects of sex steroid hormones on the growth of an aetiologic agent of chromoblastomycosis, we studied the dematiaceous fungus Phialophora verrucosa. The in vitro growth of this species on culture media containing either progesterone, testosterone or oestradiol at various concentrations was assessed. Both progesterone and testosterone inhibited the growth of P. verrucosa, whereas oestradiol did not. In other experiments, fungal cytosolic fractions were obtained and steroid binding assays were performed. These studies showed that the presence of progesterone receptors possessed two binding sites as determined by Scatchard analysis, one of which has a high affinity to progesterone (Kd = 6.02 x 10(-8) M) with total binding sites of 120 fmol micrograms-1 protein. These findings suggest that the growth of P. verrucosa is regulated by steroid hormones and that the effect of progesterone could be mediated through fungal intracellular progesterone receptors.

Laparoscopic cholecystectomy is an integral part of the general surgery operative inventory. The conventional approach may need to be modified to fit different clinical situations. We present the first reported case of a laparoscopic cholecystectomy combined with a ventral hernia repair.