Nucleos(t)ide reverse transcriptase inhibitor-sparing regimens in the era of standard 3-drug combination therapies for HIV-1 infection.

Nucleos(t)ide reverse transcriptase inhibitor (NRTI)-sparing regimens have often been selected as antiretroviral therapy (ART) for HIV-1 infection recently, but data for characteristics have been lacking. This study aimed to document the current status of NRTI-sparing regimens in the era of standard 3-drug combination therapies. We cross-sectionally compared characteristics of patients treated with NRTI-sparing regimens (NRTI-sparing group) with dolutegravir plus tenofovir alafenamide fumarate/emtricitabine as a standard ART group in 2018. The NRTI-sparing and the standard ART groups included 61 and 469 patients, respectively. The mean (± standard deviation) age and serum creatinine of the NRTI-sparing group were significantly higher than those of the standard ART group (57.6 ± 12.8 years vs 42.8 ± 10.4 years (p < 0.05) and 2.09 ± 3.10 mg/dL vs. 0.93 ± 0.19 mg/dL (p < 0.05), respectively. The percentage of patients with NRTI-sparing regimens increased with age; with less than 5% in their 50s or younger, 8.4% in their 60s, and 14.1% aged ≥ 70 years. The primary reason for switching to the NRTI-sparing regimen was due to reduced renal function. According to the limited data, viral suppression was achieved at week 48 in all patients in the NRTI-sparing group. No patient had treatment failure nor developed drug resistance. The use of NRTI-sparing regimens increased with age. They were more frequently used in patients aged ≥ 60 years and those with decreased renal function.

New Findings on the Mechanism of Perspiration Including Aquaporin-5 Water Channel.

Aquaporin-5 (AQP5) is a member of the water channel protein family. Although AQP5 has been shown to be present in sweat glands, the presence or absence of regulated intracellular translocation of AQP5 in sweat glands remains to be determined. In this article, recent findings on AQP5 in sweat glands are presented. (1) Immunoreactive AQP5 was detected in the apical membranes and the intercellular canaliculi of secretory coils, and in the basolateral membranes of the clear cells in human eccrine sweat glands. (2) AQP5 rapidly concentrated at the apical membranes during sweating in mouse sweat glands. (3) Treatment of human AQP5-expressing Madin-Darby canine kidney cells with calcium ionophore A23187 resulted in a twofold increase in the AQP5 level in the apical membranes within 5 min. (4) Anoctamin-1, a calcium-activated chloride channel was detected in the apical membranes and it completely colocalized with AQP5 in the apical membranes in mouse sweat glands. AQP5 may be involved in sweating and its translocation may help to increase the water permeability of the apical membranes of sweat glands. AQP5 is a potential target molecule for the design of a sweat-modulating drug.

Immunolocalization and translocation of aquaporin-5 water channel in sweat glands.

BACKGROUND: Aquaporin-5 (AQP5) is a member of the water channel protein family. Although AQP5 was shown to be present in sweat glands, the presence or absence of regulated intracellular translocation of AQP5 in sweat glands remained to be determined. OBJECTIVE: We investigated whether AQP5 in sweat glands translocated during sweating, and also sought to determine the intracellular signal that triggers this translocation. METHODS: Immunofluorescent analyses of AQP5 in mouse and human sweat glands were performed. Madin-Darby Canine kidney (MDCK) cell lines stably expressing human AQP5 were generated, and the regulated translocation of AQP5 in the polarized cells was assessed by immunofluorescent analysis and biotinylation assays. RESULTS: AQP5 showed rapid translocation to the apical membranes during sweating. In human eccrine sweat glands, immunoreactive AQP5 was detected in the apical membranes and the intercellular canaliculi of secretory coils, and in the basolateral membranes of the clear cells. Treatment of human AQP5-expressing MDCK cells with calcium ionophore A23187 resulted in a twofold increase of AQP5 in the apical membranes within 5min. CONCLUSION: The regulated AQP5 translocation may contribute to sweat secretion by increasing the water permeability of apical plasma membranes of sweat glands.

Separation and quantitative determination of 6α-hydroxycortisol and 6ß-hydroxycortisol in human urine by high-performance liquid chromatography with ultraviolet absorption detection.

The present study developed an high-performance liquid chromatography (HPLC) method for the simultaneous determination of urinary metabolites of endogenous cortisol, 6α-hydroxycortisol (6α-OHF) and 6ß-hydroxycortisol (6ß-OHF), in human urine, using 6α-hydroxycorticosterone as internal standard. 6α-OHF and 6ß-OHF were extracted from urine with ethyl acetate by using a Sep-Pak C(18) plus cartridge. Separation of the stereoisomers was achieved on a reversed-phase hybrid column by a gradient elution of (A) 0.05 M KH(2)PO(4)-0.01 M CH(3)COOH (pH 3.77) and (B) 0.05 M KH(2)PO(4)-0.01 M CH(3)COOH/acetonitrile (2:3, v/v). 6α-OHF and 6ß-OHF were well separated on an XTerra MS C(18) 5 µm column using two types of stepwise gradient elution program (programs 2 and 3). Resolutions of 6α-OHF and 6ß-OHF were Rs = 4.41 for program 2 and Rs = 4.60 for program 3. The analysis was performed within 23~26 min, monitored by UV absorbance at 239 nm. The lower limits of detection of 6α-OHF and 6ß-OHF were 0.80 ng per injection (s/n = ca. 8), and the lower limits of quantification were 5.02 ng/ml for 6α-OHF and 41.08 ng/ml for 6ß-OHF, respectively. The within-day reproducibilities in the amounts of 6α-OHF and 6ß-OHF determined were in good agreement with the actual amounts added, the relative errors being -5.37% and -3.73% (gradient 2) and -5.69% and -3.96% (gradient 3) for both 6α-OHF and 6ß-OHF, respectively. The inter-assay precisions (RSDs) for 6α-OHF and 6ß-OHF were less than 1.99% (gradient 2) and 2.61% (gradient 3), respectively. The present HPLC method was applied to the measurement of 6α-OHF and 6ß-OHF in urine to evaluate the time courses of 6α-hydroxylation and 6ß-hydroxylation clearances of cortisol during 40 days for phenotyping CYP3A in a healthy subject.