A prospective pilot study of circulating endothelial cells as a potential new biomarker in portal hypertension

Background/Aims: Peripheral circulating endothelial cells (CEC) have been proposed as a prognostic marker in cardiovascular diseases. Cirrhosis and portal hypertension are associated with vascular injury yet little is known about CEC count in these conditions. Therefore, we evaluated CEC count in patients with cirrhosis, and correlated it with markers of portal hypertension/disease severity. Patients/Methods: Fifteen patients with cirrhosis/portal hypertension and 15 matched controls were prospectively recruited for study participation. An automated rare cell analysis system was used to enumerate CEC from peripheral blood and correlated with clinical features. Results: Median CEC levels were significantly higher in patients with cirrhosis as compared with controls (median [interquartile range (IQR)]; cirrhosis: 73.7 cells/4 ml [53.7–140.3]; controls: 28.7 cells/4 ml [21–58.7]; P=0.021). Ratio of CEC to platelet count (CEC/PC) also distinguished patients with cirrhosis from controls (IQR; cirrhosis: 0.723 [0.396–1.672]; controls: 0.126 [0.103–0.333]; P<0.001). Receiver operator characteristic analysis revealed that CEC cut‐off of 42 cells/4 ml showed sensitivity of 87% and specificity of 74% for differentiating cirrhosis from controls (AUC: 0.74), while CEC/PC ratio at 0.21 showed sensitivity of 100% and specificity of 73% (AUC: 0.89). Furthermore, CEC/PC index was significantly elevated in patients with hepatic decompensation as defined by Child B/C (P<0.05). The intra‐ and interobserver variability correlation coefficients for CEC measurement were 0.9989 and 0.9986 respectively. Conclusion: Median CEC count and CEC/PC ratio are significantly elevated in patients with cirrhosis, with CEC/PC also increased in patients with decompensated cirrhosis. These data provide rationale for larger validation studies to assess if CEC may have prognostic utility in patients with cirrhosis and portal hypertension.     

SCALE-PWI: A pulse sequence for absolute quantitative cerebral perfusion imaging

The Bookend technique is a magnetic resonance imaging (MRI) dynamic susceptibility contrast method that provides reliable quantitative measurement of cerebral blood flow (CBF) and cerebral blood volume (CBV). The quantification is patient specific, is derived from a steady-state measurement of CBV, and is obtained from T₁ changes in the white matter and the blood pool after contrast agent injection. In the current implementation, the Bookend technique consists of three scanning steps requiring a cumulative scan time of 3 minutes 47 seconds, a well-trained technologist, and extra time for offline image reconstruction. We present an automation and acceleration of the multiscan Bookend protocol through a self-calibrating pulse sequence, namely Self-Calibrated Epi Perfusion-Weighted Imaging (SCALE-PWI). The SCALE-PWI is a single-shot echo-planar imaging pulse sequence with three modules and a total scan time of under 2 minutes. It provides the possibility of performing online, quantitative perfusion image reconstruction, which reduces the latency to obtain quantitative maps. A validation study in healthy volunteers (N=19) showed excellent agreement between SCALE-PWI and the conventional Bookend protocol (P>0.05 with Student''s t-test, r=0.95/slope=0.98 for quantitative CBF, and r=0.91/slope=0.94 for quantitative CBV). A single MRI pulse sequence for absolute quantification of cerebral perfusion has been developed.     

Advocating prevention over punishment: the risks of HIV criminalization in Burkina Faso

In 2004, parliamentarians from 12 countries in West and Central Africa created a template for legislation aimed at protecting the rights of people with HIV and stemming rising HIV infection rates by criminalizing HIV transmission. Since then, the template has been adopted as national law in 15 African countries, including Burkina Faso in 2008. The Burkina Faso law offers a number of protections for people with HIV, such as confidentiality of HIV test results, and holds the government accountable for providing health services for people with HIV and education about HIV in schools. However, other articles in the law, which criminalize HIV transmission and mandate disclosure of HIV status, may contribute to violations of the human rights of women and men with HIV. This article reviews the two cases brought in Burkina Faso under the 2008 HIV law to date, both against women, and explores the implications of specific elements of the legislation. It recommends that Burkina Faso use guidance provided by UNAIDS and the Southern Africa Development Community to repeal harmful articles in the HIV-specific legislation and implement the positive provisions. Prioritizing HIV prevention over punishment is the best way to respect the rights of people living with HIV and AIDS.     

Personalized State-space Modeling of Glucose Dynamics for Type 1 Diabetes Using Continuously Monitored Glucose,Insulin Dose,and Meal Intake: An Extended Kalman Filter Approach

An essential component of any artificial pancreas is on the prediction of blood glucose levels as a function of exogenous and endogenous perturbations such as insulin dose, meal intake, and physical activity and emotional tone under natural living conditions. In this article, we present a new data-driven state-space dynamic model with time-varying coefficients that are used to explicitly quantify the time-varying patient-specific effects of insulin dose and meal intake on blood glucose fluctuations. Using the 3-variate time series of glucose level, insulin dose, and meal intake of an individual type 1 diabetic subject, we apply an extended Kalman filter (EKF) to estimate time-varying coefficients of the patient-specific state-space model. We evaluate our empirical modeling using (1) the FDA-approved UVa/Padova simulator with 30 virtual patients and (2) clinical data of 5 type 1 diabetic patients under natural living conditions. Compared to a forgetting-factor-based recursive ARX model of the same order, the EKF model predictions have higher fit, and significantly better temporal gain and J index and thus are superior in early detection of upward and downward trends in glucose. The EKF based state-space model developed in this article is particularly suitable for model-based state-feedback control designs since the Kalman filter estimates the state variable of the glucose dynamics based on the measured glucose time series. In addition, since the model parameters are estimated in real time, this model is also suitable for adaptive control.     

The chitinase system from Trichomonas vaginalis as a potential target for antimicrobial therapy of urogenital trichomoniasis.

Chitinolytic activities in Trichomonas vaginalis membrane extracts were assessed by assays of three enzyme systems: N-acetyl-beta-D-hexosaminidase (NAHase), chitobiosidase and chitotriosidase. N-acetyl-beta-D-hexosaminidase was the enzyme that showed the highest specific activity. After successive subcutaneous inoculations into mice and parasite recovery in culture, the enzyme activities increased significantly with the number of inoculations for up to eight passages. In addition, enzyme activities were maximum at the logarithmic phase of growth. Glycol chitin, a chitinase substrate, enhanced all chitinolytic activities by about 30% and a clear-cut correlation is shown between the capacity for erythrocyte lysis by parasites and NAHase expression. Chitobiosidase and chitotriosidase activities were both inhibited at 58% and 100%, respectively, by allosamidine, a chitinase inhibitor used at 3 microM, whereas NAHase activity was not affected. Seven putative NAHase inhibitors (compounds n, 1-7), ureido and thioureido derivatives of 2-amino-2-deoxy-D-glucose were evaluated and five of them had K(i) values in the range 30-70 microM. The most active compound (compound 6) was functionally competitive with respect to the substrate with a K(i) value of 30 microM. The IC(50) values of the most active compounds on T. vaginalis were in the range 62-85 microM. These results indicate that chitinases of T. vaginalis are involved in pathogenicity and they could be an interesting target for drugs since chitinase inhibitors also inhibit parasite growth.     

Floating microspheres of cimetidine: formulation, characterization and in vitro evaluation

The present study involves preparation and evaluation of floating microspheres with cimetidine as model drug for prolongation of gastric residence time. The microspheres were prepared by the solvent evaporation method using polymers hydroxypropylmethyl cellulose and ethyl cellulose. The shape and surface morphology of prepared microspheres were characterized by optical and scanning electron microscopy, respectively. In vitro drug release studies were performed and drug release kinetics was evaluated using the linear regression method. Effects of the stirring rate during preparation, polymer concentration, solvent composition and dissolution medium on the size of microspheres and drug release were also observed. The prepared microspheres exhibited prolonged drug release (approximately 8 h) and remained buoyant for > 10 h. The mean particle size increased and the drug release rate decreased at higher polymer concentration. No significant effect of the stirring rate during preparation on drug release was observed. In vitro studies demonstrated diffusion-controlled drug release from the microspheres.     

Monoclonal antibody from vasectomized mouse identifies a conserved testis-specific antigen TSA70

Vasectomy results in the occlusion of testicular outflow, leading to autoimmunity characterized by the production of antisperm antibodies (ASA). Reports on the rise in ASA following vasectomy in several species are available; however, not much is known about the specific sperm autoantigens to which postvasectomy antibodies are directed. In the present study, monoclonal antibodies were generated using a vasectomized mouse. One of the monoclonal antibodies, D5E5, identified an approximately 70-kd antigen localized on the principal piece of the tail and also on the tip of the acrosome of mouse sperm. The cognate antigen was expressed postmeiotically in a stage-specific manner during spermiogenesis, starting from step 8 of elongating spermatids during spermiogenesis up to mature spermatozoa. The protein was conserved across the species, as observed by its presence in rat, bull, marmoset, and human sperm. Following capacitation, the antigen on the head was seen to shift to the acrosomal region and was lost after the acrosome reaction. However, the localization on tip of the acrosome still persisted, which indicates that the antigen may play a role post-acrosome reaction in sperm egg interaction. Resistance to Triton X-100 solubilization indicates that TSA70 could be an acrosomal matrix protein. In addition, we observed a significant reduction in forward progressive motility of mouse sperm treated in vitro with D5E5. In view of its testis specificity, acrosome and tail localization, and conserved nature, TSA70 is likely to play an important role in sperm function.