Denosumab for the treatment of HIV-associated osteoporosis with fractures in a premenopausal woman.

The prevalence of osteoporosis is about three times greater in people living with HIV than in the general population. Bisphosphonates are the only class of antiresorptive drugs which have proved to be safe and effective in HIV patients. However, bisphosphonates are not recommended in women of childbearing age due to an increased rate of associated neonatal complications. To the best of our knowledge no reports on the use of denosumab in HIV-infected individuals have been published so far. We describe a 38 year-old woman with HIV, osteoporosis and vertebral fractures treated with denosumab, a monoclonal antibody targeting RANKL. After four years of treatment, bone mineral density improved, no new fractures occurred, and neither HIV reactivation nor opportunistic infections were observed. We show that denosumab could be a safe and effective approach for osteoporosis in patients with HIV and could be considered in women of childbearing age.

Intestinal Alkaline Phosphatase Attenuates Alcohol-Induced Hepatosteatosis in Mice.

BACKGROUND AND AIMS: Bacterially derived factors from the gut play a major role in the activation of inflammatory pathways in the liver and in the pathogenesis of alcoholic liver disease. The intestinal brush-border enzyme intestinal alkaline phosphatase (IAP) detoxifies a variety of bacterial pro-inflammatory factors and also functions to preserve gut barrier function. The aim of this study was to investigate whether oral IAP supplementation could protect against alcohol-induced liver disease. METHODS: Mice underwent acute binge or chronic ethanol exposure to induce alcoholic liver injury and steatosis ± IAP supplementation. Liver tissue was assessed for biochemical, inflammatory, and histopathological changes. An ex vivo co-culture system was used to examine the effects of alcohol and IAP treatment in regard to the activation of hepatic stellate cells and their role in the development of alcoholic liver disease. RESULTS: Pretreatment with IAP resulted in significantly lower serum alanine aminotransferase compared to the ethanol alone group in the acute binge model. IAP treatment attenuated the development of alcohol-induced fatty liver, lowered hepatic pro-inflammatory cytokine and serum LPS levels, and prevented alcohol-induced gut barrier dysfunction. Finally, IAP ameliorated the activation of hepatic stellate cells and prevented their lipogenic effect on hepatocytes. CONCLUSIONS: IAP treatment protected mice from alcohol-induced hepatotoxicity and steatosis. Oral IAP supplementation could represent a novel therapy to prevent alcoholic-related liver disease in humans.

Clinical pharmacology of tenofovir clearance: a pharmacokinetic/pharmacogenetic study on plasma and urines.

The HIV virus and hepatitis B virus nucleotide reverse transcriptase inhibitor tenofovir has been associated with proximal tubular toxicity; the latter was found to be predicted by plasma concentrations and with single-nucleotide polymorphisms in transporters-encoding genes. A cross-sectional analysis in adult HIV-positive patients with estimated creatinine clearance >60 ml min-1 was performed. Twelve-hour plasma and urinary tenofovir concentrations and single-nucleotide polymorphisms in several transporter-encoding genes were analysed. In 289 patients 12-h tenofovir plasma, urinary and urinary to plasma ratios were 69 ng ml-1 (interquartile range 51.5-95), 24.3 mg ml-1 (14.3-37.7) and 384 (209-560). At multivariate analysis estimated creatinine clearance, protease inhibitors co-administration and SLC28A2 CT/TT genotypes were independently associated with plasma tenofovir exposure; ABCC10 GA/AA genotypes and protease inhibitor co-administration were independently associated with the urinary to plasma tenofovir ratio. Tenofovir clearance was associated with genetic polymorphisms in host genes and with co-administered drugs: if confirmed by ongoing studies these data may inform treatment tailoring and/or dose reductions.

Intestinal alkaline phosphatase prevents metabolic syndrome in mice.

Metabolic syndrome comprises a cluster of related disorders that includes obesity, glucose intolerance, insulin resistance, dyslipidemia, and fatty liver. Recently, gut-derived chronic endotoxemia has been identified as a primary mediator for triggering the low-grade inflammation responsible for the development of metabolic syndrome. In the present study we examined the role of the small intestinal brush-border enzyme, intestinal alkaline phosphatase (IAP), in preventing a high-fat-diet-induced metabolic syndrome in mice. We found that both endogenous and orally supplemented IAP inhibits absorption of endotoxin (lipopolysaccharides) that occurs with dietary fat, and oral IAP supplementation prevents as well as reverses metabolic syndrome. Furthermore, IAP supplementation improves the lipid profile in mice fed a standard, low-fat chow diet. These results point to a potentially unique therapy against metabolic syndrome in at-risk humans.

Cross sectional study investigating the differences in knowledge and behaviors about HPV between vaccinated and non-vaccinated girls.

INTRODUCTION: The aim of the presents study was to compare the level of knowledge about Human Papilloma Virus (HPV) in vaccinated and non-vaccinated girls and to highlight the reasons why non-vaccinated girls refuse vaccination. METHODS: A cross-sectional study was conducted from October 2012 to June 2013 in Turin (Piemonte Region, Italy). Questionnaires were administered to girls attending secondary and high schools randomly selected. RESULTS: A total of 576 were compiled. The principle sources of information were parents and health workers. The main reported reasons for non-adherence to vaccination were the disagreement of the parents among the 11-12 years group (45.3%) and the lack of evidence on efficacy among the 18 years group (26.8%). By comparing the level of knowledge there was a statistically significant difference between groups: vaccinated girls reported higher score than the unvaccinated group in several questions (p ≤ 0.05). CONCLUSIONS: Our findings show a lack of information about HPV infection. Parents, school and health care workers have a central role in girl's education and choices about HPV vaccination. The communication campaign for the prevention of cervical cancer must therefore be characterised by messages able to clarify and consolidate messages that may have been partially received or misunderstood.

Intestinal alkaline phosphatase promotes gut bacterial growth by reducing the concentration of luminal nucleotide triphosphates.

The intestinal microbiota plays a pivotal role in maintaining human health and well-being. Previously, we have shown that mice deficient in the brush-border enzyme intestinal alkaline phosphatase (IAP) suffer from dysbiosis and that oral IAP supplementation normalizes the gut flora. Here we aimed to decipher the molecular mechanism by which IAP promotes bacterial growth. We used an isolated mouse intestinal loop model to directly examine the effect of exogenous IAP on the growth of specific intestinal bacterial species. We studied the effects of various IAP targets on the growth of stool aerobic and anaerobic bacteria as well as on a few specific gut organisms. We determined the effects of ATP and other nucleotides on bacterial growth. Furthermore, we examined the effects of IAP on reversing the inhibitory effects of nucleotides on bacterial growth. We have confirmed that local IAP bioactivity creates a luminal environment that promotes the growth of a wide range of commensal organisms. IAP promotes the growth of stool aerobic and anaerobic bacteria and appears to exert its growth promoting effects by inactivating (dephosphorylating) luminal ATP and other luminal nucleotide triphosphates. We observed that compared with wild-type mice, IAP-knockout mice have more ATP in their luminal contents, and exogenous IAP can reverse the ATP-mediated inhibition of bacterial growth in the isolated intestinal loop. In conclusion, IAP appears to promote the growth of intestinal commensal bacteria by inhibiting the concentration of luminal nucleotide triphosphates.

A novel approach to maintain gut mucosal integrity using an oral enzyme supplement.

OBJECTIVE: To determine the role of intestinal alkaline phosphatase (IAP) in enteral starvation-induced gut barrier dysfunction and to study its therapeutic effect as a supplement to prevent gut-derived sepsis. BACKGROUND: Critically ill patients are at increased risk for systemic sepsis and, in some cases, multiorgan failure leading to death. Years ago, the gut was identified as a major source for this systemic sepsis syndrome. Previously, we have shown that IAP detoxifies bacterial toxins, prevents endotoxemia, and preserves intestinal microbiotal homeostasis. METHODS: WT and IAP-KO mice were used to examine gut barrier function and tight junction protein levels during 48-hour starvation and fed states. Human ileal fluid samples were collected from 20 patients postileostomy and IAP levels were compared between fasted and fed states. To study the effect of IAP supplementation on starvation-induced gut barrier dysfunction, WT mice were fasted for 48 hours +/- IAP supplementation in the drinking water. RESULTS: The loss of IAP expression is associated with decreased expression of intestinal junctional proteins and impaired barrier function. For the first time, we demonstrate that IAP expression is also decreased in humans who are deprived of enteral feeding. Finally, our data demonstrate that IAP supplementation reverses the gut barrier dysfunction and tight junction protein losses due to a lack of enteral feeding. CONCLUSIONS: IAP is a major regulator of gut mucosal permeability and is able to ameliorate starvation-induced gut barrier dysfunction. Enteral IAP supplementation may represent a novel approach to maintain bowel integrity in critically ill patients.

Intestinal alkaline phosphatase inhibits the proinflammatory nucleotide uridine diphosphate.

Uridine diphosphate (UDP) is a proinflammatory nucleotide implicated in inflammatory bowel disease. Intestinal alkaline phosphatase (IAP) is a gut mucosal defense factor capable of inhibiting intestinal inflammation. We used the malachite green assay to show that IAP dephosphorylates UDP. To study the anti-inflammatory effect of IAP, UDP or other proinflammatory ligands (LPS, flagellin, Pam3Cys, or TNF-α) in the presence or absence of IAP were applied to cell cultures, and IL-8 was measured. UDP caused dose-dependent increase in IL-8 release by immune cells and two gut epithelial cell lines, and IAP treatment abrogated IL-8 release. Costimulation with UDP and other inflammatory ligands resulted in a synergistic increase in IL-8 release, which was prevented by IAP treatment. In vivo, UDP in the presence or absence of IAP was instilled into a small intestinal loop model in wild-type and IAP-knockout mice. Luminal contents were applied to cell culture, and cytokine levels were measured in culture supernatant and intestinal tissue. UDP-treated luminal contents induced more inflammation on target cells, with a greater inflammatory response to contents from IAP-KO mice treated with UDP than from WT mice. Additionally, UDP treatment increased TNF-α levels in intestinal tissue of IAP-KO mice, and cotreatment with IAP reduced inflammation to control levels. Taken together, these studies show that IAP prevents inflammation caused by UDP alone and in combination with other ligands, and the anti-inflammatory effect of IAP against UDP persists in mouse small intestine. The benefits of IAP in intestinal disease may be partly due to inhibition of the proinflammatory activity of UDP.

[The influence of an air ionizer on the air ionic composition].

The air ionic composition in a classroom was determined during the operation of a Neo Tec XJ-2100 ionizer (Germany). The amount of ions in the air was measured before and after the switch-on of the ionizer. It was shown to vary depending on the mode of operation of the device and differ from the recommended normal values. The necessity of checking up the work of air ionizers both in living quarters and at workplaces is discussed with the application of air ion counters making it possible to carry out monitoring of the air ionic composition and estimate its compliance with the sanitary and hygienic norms.

Epidemiology and microbiology of prosthetic joint infections: a nine-year, single-center experience in Pavia, Northern Italy.

BACKGROUND: Prosthetic joint infections (PJIs) are a growing matter of concern due to their economic and social burden on health systems. In Italy, surgical data on PJIs are available in a national registry, but microbiological data are still scarce. MATERIALS AND METHODS: We performed a retrospective study at a single center with records of patients treated for primary PJIs of knee or hip from January 1, 2011, to May 30, 2018. Patients with infections of osteosynthesis means and external devices were excluded, as well as PJI recurrences and polytrauma patients. Infections were diagnosed according to IDSA and MSIS criteria. We collected data on demographics, risk factors and microbiology. All patients seen at our center undergo blood cultures and synovial fluid cultures, periarticular biopsy and prosthesis sonication by Bactosonic®. This was used only after 2014. Bacterial identification is achieved by MALDI-TOF, PHOENIX 100 and standard methods. Chi-square or Fisher tests were used to test statistical differences in proportions. RESULTS: Fifty-one patients matched our inclusion criteria. Of these, 16 (31.4%) were enrolled before 2014. The median age was 68.5 (range 22-88). The most common risk factors were obesity (34%), diabetes (21%) and chronic kidney disease (14%). Seventeen patients were diagnosed with a culture-negative PJIs (33.3%). Staphylococcus aureus was the most commonly isolated pathogen (14/51, 27.5%), followed by coagulase-negative staphylococci (7/51, 13.7%). Methicillin-resistant S. aureus rate was 28.6%. The rate of culture-negative PJIs dropped from 56 to 22% after 2014, with a significant difference between the two time periods (p = 0.016). CONCLUSIONS: The introduction of sonication dramatically increased our diagnostic accuracy. Our microbiological data are in line with those from other studies conducted in Italy.

Hepatitis B Infection, Viral Load and Resistance in HIV-Infected Patients in Mozambique and Zambia.

BACKGROUND: Few data on the virological determinants of hepatitis B virus (HBV) infection are available from southern Africa. METHODS: We enrolled consecutive HIV-infected adult patients initiating antiretroviral therapy (ART) at two urban clinics in Zambia and four rural clinics in Northern Mozambique between May 2013 and August 2014. HBsAg screening was performed using the Determine® rapid test. Quantitative real-time PCR and HBV sequencing were performed in HBsAg-positive patients. Risk factors for HBV infection were evaluated using Chi-square and Mann-Whitney tests and associations between baseline characteristics and high level HBV replication explored in multivariable logistic regression. RESULTS: Seventy-eight of 1,032 participants in Mozambique (7.6%, 95% confidence interval [CI]: 6.1-9.3) and 90 of 797 in Zambia (11.3%, 95% CI: 9.3-13.4) were HBsAg-positive. HBsAg-positive individuals were less likely to be female compared to HBsAg-negative ones (52.3% vs. 66.1%, p<0.001). Among 156 (92.9%) HBsAg-positive patients with an available measurement, median HBV viral load was 13,645 IU/mL (interquartile range: 192-8,617,488 IU/mL) and 77 (49.4%) had high values (>20,000 UI/mL). HBsAg-positive individuals had higher levels of ALT and AST compared to HBsAg-negative ones (both p<0.001). In multivariable analyses, male sex (adjusted odds ratio: 2.59, 95% CI: 1.22-5.53) and CD4 cell count below 200/µl (2.58, 1.20-5.54) were associated with high HBV DNA. HBV genotypes A1 (58.8%) and E (38.2%) were most prevalent. Four patients had probable resistance to lamivudine and/or entecavir. CONCLUSION: One half of HBsAg-positive patients demonstrated high HBV viremia, supporting the early initiation of tenofovir-containing ART in HIV/HBV-coinfected adults.

WHO Neuropsychiatric AIDS study, cross-sectional phase II. Neuropsychological and neurological findings.

BACKGROUND: The neuropsychological and neurological complications of HIV-1 infection and AIDS were explored within the cross-sectional phase of the WHO Neuropsychiatric AIDS Study. Special attention was devoted to the controversial issue of the prevalence and clinical significance of subtle cognitive deficits in asymptomatic seropositive subjects. METHODS: A neuropsychological test battery validated for cross-cultural use, a structured interview for the diagnosis of dementia, a rating scale of functioning in daily living activities, and a neurological module were administered to representative samples of seropositive subjects and to matched seronegative controls living in the five geographic areas predominantly affected by the HIV-1 epidemic. Data are available for five centers. RESULTS: The prevalence of global neuropsychological impairment was significantly increased in asymptomatic seropositive subjects compared with controls in only two centers. A significant effect of education on neuropsychological performance was observed among asymptomatic seropositive individuals. In the two African centers, low-education, but not high-education, asymptomatic seropositive persons had an impaired performance. The frequency of impaired functioning in daily living activities and of neurologic abnormalities was higher in symptomatic, but not in asymptomatic, seropositive subjects compared with controls in all centers. CONCLUSIONS: These data suggest that the risk of subtle cognitive deficits may be increased in asymptomatic stages of HIV-1 infection. However, these deficits are not associated with neurologic changes and do not seem to affect subjects' social functioning.

Neurological complications of HIV-1-seropositive internal medicine inpatients in Kinshasa, Zaire.

Because little was known about the prevalence of neurological complications of human immunodeficiency virus type 1 (HIV-1) infection in Africa, we conducted a cross-sectional study among consecutive admissions to the internal medicine wards of Mama Yemo Hospital in Kinshasa, Zaire. Of the 196 patients studied, 104 (53%) were HIV-1 seropositive, of whom 50 (48%) had stage 3 and 49 (47%) had stage 4 HIV-1 infection according to the provisional WHO staging criteria for HIV infection. Neuropsychiatric abnormalities were present in 43 (41%) of 104 HIV-1-seropositive patients. Of the HIV-1-seropositive patients, 9 (8.7%; 95% confidence interval, 4-16%) were diagnosed as having possible HIV-1-associated dementia complex, 1 (1%) as having possible HIV-1 myelopathy, and 3 (2.7%) as having possible HIV-1-associated minor cognitive/motor disorder. Definitive diagnoses could not be made because there were no facilities for neuroimaging and neuropathology. Meningitis caused by cryptococcus was diagnosed in six (5.6%) and by Mycobacterium avium in two (2%) of the HIV-1 seropositive patients. Acute onset hemiplegia, believed to be due to stroke, was present in four (4%) of the HIV-1-seropositive patients. The prevalence of other central nervous system opportunistic infections and mass lesions, especially toxoplasmic encephalitis, could not be assessed. In this population of Zairian inpatients, the prevalence of neurological complications of HIV-1 infection was similar to that observed in industrialized countries among patients with advanced HIV disease.

[HIV-1 cognitive and motor syndrome]. / Syndrome cognitif et moteur associé au VIH-1.

The central nervous system (CNS) is often affected by HIV-1 infection. Over 40% of AIDS cases present with neurological symptoms and CNS lesion are detected by anatomical and pathological studies in 80 to 90% of AIDS cases. There may be infections and tumors secondary to the immunodeficiency and pathologies may occur directly due to the neurotropism of the virus. Neurological problems associated with HIV-infection include encephalopathies, myelopathies, neuropathies and myopathies. HIV-1-induced encephalopathy may develop at any stages of HIV-1 infection and affects all risk groups equally. Its frequency worldwide is between 4 and 65% among individuals seropositive for HIV-1. The frequencies reported differ between studies due to differences in sampling methods, geographical factors, diagnostic criteria and investigative methods used. The pathogenesis of HIV-1-associated encephalopathy is not understood, but there are several hypotheses. The involvement of HIV-1 infected macrophages and microglial cells has been demonstrated. Indirect mechanisms such as release of lymphokines (tumor necrosis factor-TNF alpha- and interleukin-1) and neurotoxicity of the HIV envelope protein, gp 120, have also been suggested. This disorder is known as HIV-1-associated cognitive and motor syndrome. It presents clinically as a form of sub-cortical dementia with cognitive problems, motor deficits and behavioral disorders depending on the type and stage of HIV infection. The diagnosis can only be made after all other infections and tumors common in HIV-1 patients have been ruled out by appropriate investigations such as cerebrospinal fluid analysis, cerebral scan and magnetic resonance imaging. Electrophysiological studies, such as evoked responses and electroencephalograms, are particularly useful in its diagnosis. Anatomical examination shows diffuse paleness of the white matter, multi-nucleated giant cells and microglial nodes. Neuropsychological studies could be of value in diagnosis and in assessing the response to anti-retroviral treatment. There is currently no specific therapy for HIV-1-associated cognitive and motor syndrome. The use of new nucleoside analogue drugs in combination with existing drugs may provide new approaches to managing these patients.

[Correction of extensive defects of long bones by using bi- and multifocal distraction-compression osteosynthesis]. / Zameshchenie obshirnykh defektov dlinnykh kostei s pomoshch'iu bi- i polilokal'nogo distraktsionno-kompressionnogo osteosinteza.

Replacement of extensive defects in long bones on the basis of one or more osteotomies is analysed. The authors substantiate the principles and the new methods of +multifocal distraction-compression osteosynthesis which allow healing of the fracture to be combined with accelerated replacement of the bone defect. The errors, complications, and the results of treatment of 104 patients are discussed to evaluate the efficacy of the applied methods and confirm the drawn conclusions. Particular attention is paid to the significance of a complex of optimal mechanical and biological factors in the formation of the regenerative tissue in accelerated replacement of the bone defects on the basis of several osteotomies. It is shown that the advantage of the new variants of the methods of replacement of long tubular bones on the basis of several osteotomies consists in combination of the healing of open fractures with accelerated restoration of the length of the extremity.

[Expediency of plastic surgery methods with osteomyelitic cavities]. / O tselesoobraznosti nekotorykh sposobov plastiki osteomieliticheskikh polostei.

The fitness of plaster filling, Schede's method, muscular pedicled autograft and of 3 biopolymeric compositions with the use of ultrasound for the grafting of bone cavities was tested in 206 experiments on dogs. The basic possibility of bone cavity filling with a biopolymeric composition was demonstrated.

Recyclable single-stranded DNA template for synthesis of siRNAs.

RNA interference is a post-transcriptional silencing mechanism triggered by the bioavailability and/or exogenous introduction of double-stranded RNA (dsRNA) into cells. Here we describe a novel method for the synthesis of siRNA in a single vessel. The method employs in vitro transcription and a single-stranded DNA (ssDNA) template and design, which incorporates upon self-annealing, two promoters, two templates, and three loop regions. Using this method of synthesis we generated efficacious siRNAs designed to silence both exogenous and endogenous genes in mammalian cells. Due to its unique design the single-stranded template is easily amenable to adaptation for attachment to surface platforms for synthesis of siRNAs. A siRNA synthesis platform was generated using a 3' end-biotinylated ssDNA template tethered to a streptavidin coated surface that generates stable siRNAs under multiple cycles of production. Together these data demonstrate a unique and robust method for scalable siRNA synthesis with potential application in RNAi-based array systems.