The Fourth Major Restriction Factor Against HIV/SIV

Human and simian immunodeficiency viruses (HIV/SIVs) carry a unique set of accessory proteins that enhance virus replication in an optimized manner. These viral proteins specific to HIV/SIVs are designated Vif, Vpx, Vpr, Vpu, and Nef, and are functional in certain cell types (Malim and Emerman, 2008; Fujita et al., 2010). While viruses of the HIV-1 group do not encode Vpx, the other HIV-2/SIVs are unable to replicate in cells of the myeloid lineage such as monocyte-derived dendritic cells (MDDCs) and macrophages (MDMs) in the absence of Vpx (Fujita et al., 2010). Vpx and its structural close relative Vpr are leas

Morphological study on biologically distinct vpx/vpr mutants of HIV-2

We have previously shown that human immunodeficiency virus type 2 (HIV-2) without functional vpx and vpr genes is severely defective for viral growth in lymphocytic cells, and suggested that the virions produced in the absence of Vpx and Vpr are critically damaged. To examine the nature of replication-defect for the vpx/vpr double mutant, we quantitatively and morphologically studied the virions produced in cells transfected or infected with wild type clone, single (vpx and vpr mutants) or the double mutant. While no significant difference in virion production was found for various virus clones in transfected cells, a major growth retardation in infected cells was readily observed for the vpx and vpx/vpr mutants. In particular, no viral growth was detected for the double mutant. By contrast to the very distinct growth characteristics of the three mutant clones, no appreciable difference in virion morphology was noted. These results indicated that Vpx and Vpr of HIV-2 may cooperatively contribute to virion infectivity without affecting virion morphogenesis

Manifestation of Headache Affecting Quality of Life in Long COVID Patients

Objectives: The present study aimed to elucidate the characteristics of long COVID patients with headaches. Methods: A single-center retrospective observational study was performed for long COVID outpatients who visited our hospital from 12 February 2021 to 30 November 2022. Results: A total of 482 long COVID patients, after excluding 6, were divided into two groups: the Headache group of patients with complaints of headache (113 patients: 23.4%) and the remaining Headache-free group. Patients in the Headache group were younger (median age: 37 years) than patients in the Headache-free group (42 years), while the ratio of females (56%) in the Headache group was nearly the same as that in the Headache-free group (54%). The proportion of patients in the Headache group who were infected in the Omicron-dominant phase (61%) was larger than the proportions of patients infected in the Delta (24%) and preceding (15%) phases, and that trend was significantly different from the trend in the Headache-free group. The duration before the first visit for long COVID was shorter in the Headache group (71 days) than in the Headache-free group (84 days). The proportions of patients in the Headache group with comorbid symptoms, including general fatigue (76.1%), insomnia (36.3%), dizziness (16.8%), fever (9.7%), and chest pain (5.3%) were larger than the proportions of patients in the Headache-free group, whereas blood biochemical data were not significantly different between the two groups. Interestingly, patients in the Headache group had significant deteriorations of scores indicating depression and scores for quality of life and general fatigue. In multivariate analysis, headache, insomnia, dizziness, lethargy, and numbness were shown to be involved in the quality of life (QOL) of long COVID patients. Conclusions: The manifestation of headaches related to long COVID was found to have a significant impact on social and psychological activities. Alleviation of headaches should be a priority for the effective treatment of long COVID

Generation and characterization of APOBEC3G-positive 293T cells for HIV-1 Vif study

We have established a number of 293T cell lines that express a human anti HIV-1 factor APOBEC3G. Out of seven cell clones examined, four were readily demonstrated to express APOBEC3G by immunoblotting analysis. In particular, two clones (A3G-C1 and -C4) were found to produce a much higher level of functional APOBEC3G relative to that by pooled cell clones. The transfection efficiency of all these cell clones were similar to that of the parental cells, producing a comparable level of virions upon transfection of wild type and vif -minus proviral DNA clones. Furthermore, the expression level of APOBEC3G in the best cell line (A3G-C1) was far much higher than those of an APOBEC3G-positive lymphocyte cell line and peripheral blood mononuclear cells. We finally monitored the incorporation of APOBEC3G into virions produced in A3G-C1. APOBEC3G was easily detected in progeny viral particles upon transfection of vif -minus proviral clone but not of wild type. These results indicated that our new A3G-C1 cell line is eminently useful for various studies on the interaction of human APOBEC3G and HIV-1 Vif

Gender-Dependent Characteristics of Serum 1,25-Dihydroxyvitamin D/25-Hydroxyvitamin D Ratio for the Assessment of Bone Metabolism

Y Objectives Vitamin D deficiency, which is common worldwide, increases the risks of falls and fractures and can lead to increased morbidity and mortality. However, the clinical utility and relevance of vitamin D activation remain unknown. The aim of the present study was to clarify the clinical usefulness of serum 1,25-dihydroxyvitamin D (1,25D)/25-hydroxyvitamin D (25D) ratio for assessment of the extent of bone metabolism. Methods We retrospectively screened data for 87 patients whose serum 1,25D and 25D levels were measured. Eight patients who were taking vitamin D preparations were excluded, and data for 79 patients (33 males and 46 females) were analyzed. Since menopausal status can be associated with serum vitamin D level, we divided the patients by gender and divided the female patients into two groups at the age of 50 years. Results The median serum 1,25D/25D ratio was significantly lower in males than in females, with the most considerable difference in all males [4.1 (interquartile range: 2.3-5.8) x 10(-3)] versus elderly females (aged >= 50 years) [7.9 (3.3-10.1) x 10(-3)). Main disorders were endocrine (30.6%), inflammatory (18.5%), and bone related (16.7%) disorders. The ratios of serum 1,25D/25D had significant negative correlations with femoral dual-energy X-ray absorptiometry % young adult mean (DEXA %YAM) (R=-0.35) and lumbar DEXA %YAM (R=-0.32). Significant correlations were found between the 1,25D/25D ratio and serum levels of inorganic phosphate (iP), parathyroid hormone, and alkaline phosphatase (ALP). The 1,25D/25D ratio had gender specific characteristics: the ratio was significantly correlated with age in males (R=-0.49), while it was significantly correlated with BMI in females (R=0.34). Conclusions The results of this study suggested that vitamin D activity is negatively correlated with bone mineral density, being reduced in aged males but enhanced in obese females

Treatment outcomes of laparoscopic radical prostatectomy at Kawasaki Medical School Hospital

　Laparoscopic radical prostatectomy (LRP) was carried out in 196 patients with prostate cancer between December 2009 and November 2017 at Kawasaki Medical School Hospital, and the therapeutic outcomes were assessed. An extraperitoneal approach was used in all cases except 1 and the median follow-up period was 55 months (range, 10-117 months). The median patient age was 69 years (range, 56-79 years), median body mass index was 23.3 kg/m2 (range, 15.2-33.2 kg/m2 ), and median prostate-specific antigen (PSA) level at diagnosis was 7.4 ng/mL (range, 2.2-42.0 ng/mL). Clinical stages of T1c, T2a, T2b, T2c, T3a, and T3b accounted for 63, 43, 31, 57, 1, and 1 case, respectively, while Gleason scores at biopsy of ≥ 6, 7, and ≥ 8 accounted for 26, 138, and 32 cases, respectively. The median prostate volume was 22.0 mL (range, 7.3-65.6 mL), median operating time was 266 minutes (range, 142-540 minutes), and median blood loss (including in urine) was 650 mL (range, 10-5,800 mL). During the initial induction period, 94 patients received autologous blood transfusion and 7 received allogeneic blood transfusion. Nerve-sparing prostatectomy was performed in 17 cases (bilateral in 3, unilateral in 14). Capsular invasion was observed in 57 cases (29.1%) and positive resection margins were observed in 51 cases (26.4%). The median indwelling catheter duration was 6 days (range, 4-26 days) and the median hospital stay after surgery was 11 days (range, 8-34 days). The main complications were intraoperative rectal injury in 7 cases (3.6%), postoperative inguinal hernia in 28 (14.3%), and urethral stenosis in 8 (4.1%). The rate of urinary incontinence at ≥ 1 year after surgery was 32.7% and the rate of PSA recurrence was 15.8%. The overall survival rate was 95.6% at 5 years and 94.7% at 10 years. In conclusion, the oncological outcomes were similar to that reported by previous reports, but postoperative stress urinary incontinence and complications were slightly worse. In the future, further improvement of the surgical technique was desired

Outcomes of robot-assisted partial nephrectomy in the treatment of renal cell carcinoma at Kawasaki Medical School Hospital

　Robot-assisted partial nephrectomy (RAPN) was introduced in our hospital for treating small renal cell carcinoma in May 2018; we examined treatment outcomes in 24 patients (25 kidneys) who had undergone this procedure till 2019. The median observation period was 11 months (range, 1-17 months). The patients’ age range was 43-77 years (median, 68 years). Fourteen men and 10 women underwent the procedure. Their BMI was 17.9-39.7 (median, 24.1) kg/m2 . In one patient, RAPN was performed twice at different times for treating bilateral renal cancer. The right kidney was affected in 12 cases and the left kidney in 13 cases. The clinical cancer stage was T1a in 20 cases and T1b in 5 cases. Tumor sizes were 0.9-6.2 cm (median, 2.5 cm), and RENAL nephrometry scores were 4-10 (median, 7). The transperitoneal approach was used in 22 cases, and the retroperitoneal approach in 3. The operating durations were 147-358 min (median, 225 min), console durations were 59-394 min (median, 152 min), and renal ischemia durations were 8-54 min (median, 21 min). Blood loss was 10-700 ml (median 10 ml), and none of the patients underwent blood transfusion. The histopathological analysis of the resected tumors revealed clear cell renal cell carcinoma in 20 cases, chromophobe renal cell carcinoma in 2 cases, and papillary renal cell carcinoma, angiomyolipoma, and leiomyoma in 1 case each. All margins were negative. The postoperative hospital stay lengths were 5-14 days (median, 9 days). The postoperative deterioration in renal function was mild, and there were no severe complications. In the early stages after its introduction, RAPN was safely performed and allowed for the preservation of renal function. We plan to continue studying more cases going forward

〔研究ノート〕慢性腎臓病発症初期ラットの腎障害抑制および骨密度維持に対する大豆イソフラボン抽出物あるいはアルギニンの有効性評価に関する基礎研究

The objective of the present study was to elucidate the efficacy of soybean isoflavones extracted from hypocotyl and L-Arginine on the renal function, bone mineral density, and bone strength of adult model rats in the relatively early stage of chronic kidney disease(CKD). Based on our previous study, 360 mg/kg body weight of adenine suspended in a 2% methyl cellulose solution was administered intragastrically for six days to generate CKD model rats. For the intact/control condition, a 2% methyl cellulose solution was administered to the rats for six days. After the administration of adenine, 32 rats were randomly divided into four groups, and each group was fed a diet containing 20% casein protein(20CA, as a control, Group CA), 20CA+0.20% soy isoflavone extract(Group IF), 20CA+1.0% L-Arginine(Group Arg), and 20CA+0.20% soy isoflavone extract and 1.0% L-Arginine(IFA)at 15 g/day for 25 days, respectively. Intact rats were also fed the control diet as in group CA. Group CA had apparent renal failure, indicated by swelling of the kidneys, a decrease in creatinine clearance(CCr), increased BUN and proteinuria and lower hematocrit, as well as bone loss estimated by a lower bone weight, femoral-BMD and bone strength than the intact control group. These results showed that adenine administration results in the development of CKD and bone loss under the conditions used in the present study. The results were as follows: Compared with group CA, 1)group IF showed a significantly higher cancellous BMD in the femur, but no significant differences were observed in the cortical BMD, bone strength, kidney weight, CCr, urinary protein excretion or urinary NAG activity. 2)group Arg showed no significant differences in any parameters related to the renal function, BMD or bone strength. 3)group IFA showed significantly lower values for proteinuria, higher values for the urinary NAG activity and higher values for the femoral cancellous BMD. However, there were no significant differences observed in the kidney weight, CCr, bone weight or cortical BMD. Based on these results, we concluded that the dietary soy isoflavone extract and L-Arginine maintained the bone BMD, albeit to a limited extent, and suppressed the renal failure of the rats in the early stage of CKD under the conditions used in the present study

Metal-Chelating Inhibitors of a Zinc Finger Protein HIV-EP1. Remarkable Potentiation of Inhibitory Activity by Introduction of SH Groups (BIOORGANIC CHEMISTRY-Bioactive Chemistry)

HIV-EP1 is a C2H2 type zinc finger protein which binds to DNA kB site present in the long terminal repeat of HIV provirus. Previously we have reported zinc chelators having histidine-pyridine-histidine skeleton and were successful to inhibit the DNA binding of HIV-EP1 by removing zinc from the zinc finger domain. Aiming at the potentiation of the inhibitory activity, we synthesized novel chelators comprising pyridine and aminoalkylthiol. These showed marked inhibitory activity on the DNA binding of HIV-EP1. In particular, one of them having bis(2-mercaptoethyl)amino side chain showed inhibitory activity (IC50, ~4mM), 10 times stronger compared with the strongest inhibitor that we reported previously