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BACKGROUND: Vitamin D insufficiency plays an important role in the development of fibrosis in chronic liver disease. METHODS: This was a cross sectional study from Thailand. Liver fibrosis was assessed by transient elastography. Serum 25 hydroxy vitamin D[25(OH)D] 14 kPa. Median (IQR) duration on TDF was 5(4-7) years. The median eGFR was 96.9 ml/min/1.732. The median (IQR) serum 25(OH)D levels prior to and following TDF were 24.8(21.3-30.6) ng/ml, and 22.8(18.0-27.7) ng/ml, respectively; p=<0.001). The proportion of patients with hypovitaminosis D significantly increased from 72.2% (95% confidential interval:95%CI: 64.7-78.6) prior to TDF to 84.2% (95% CI: 77.7-89.0) after taking TDF (p=0.01). Factors associated with hypovitaminosis D by multivariate analysis were female sex [adjusted odd ratio, OR3.8 (95% CI1.1-13.7), p=0.038]; and duration of ART >5 years [OR 3.3(95% CI 1.2-8.8), p=0.017]. Vitamin D levels were not associated with significant liver fibrosis. CONCLUSIONS: Although our HIV/HBV co-infected patients live in the tropics, there was a high prevalence of hypovitaminosis D, especially in female patients and those receiving prolonged ART. Since HIV/HBV co-infection requires long term use of the HBV-active drug, TDF that can also contribute to bone loss, routine vitamin D assessment and supplementation as necessary should be considered.