Investigation of red blood cell antigens with highly fluorescent and stable semiconductor quantum dots

We report a new methodology for red blood cell antigen expression determination by a simple labeling procedure employing luminescent semiconductor quantum dots. Highly luminescent and stable core shell cadmium sulfide/cadmium hydroxide colloidal particles are obtained, with a predominant size of 9 nm. The core-shell quantum dots are functionalized with glutaraldehyde and conjugated to a monoclonal anti-A antibody to target antigen-A in red blood cell membranes. Erythrocyte samples of blood groups A+, A2+, and O+ are used for this purpose. Confocal microscopy images show that after 30 min of conjugation time, type A+ and A2+ erythrocytes present bright emission, whereas the O+ group cells show no emission. Fluorescence intensity maps show different antigen expressions for the distinct erythrocyte types. The results obtained strongly suggest that this simple labeling procedure may be employed as an efficient tool to investigate quantitatively the distribution and expression of antigens in red blood cell membranes.     

Identification of a novel receptor-like protein kinase that interacts with a geminivirus nuclear shuttle protein

Despite extensive studies in plant virus-host interactions, the molecular mechanisms of geminivirus movement and interactions with host components remain largely unknown. A tomato kinase protein and its soybean homolog were found to interact specifically with the nuclear shuttle protein (NSP) of Tomato golden mosaic virus (TGMV) and Tomato crinkle leaf yellows virus (TCrLYV) through yeast two-hybrid screening and in vitro protein binding assays. These proteins, designated LeNIK (Lycopersicon esculentum NSP-Interacting Kinase) and GmNIK (Glycine max NIK), belong to the LRR-RLK (leucine rich-repeat receptor-like kinase) family that is involved in plant developmental processes and/or resistance response. As such, NIK is structurally organized into characteristic domains, including a serine/threonine kinase domain with a nucleotide binding site at the C-terminal region, an internal transmembrane segment and leucine-rich repeats (LRR) at the N-terminal portion. The potential significance of the NSP-NIK interaction is discussed.     

The myenteric plexus of the rat colon after fecal stream diversion: a morpho-quantitative study

The myenteric plexus undergoes adaptive changes under several conditions. Mucosal and muscular alterations of the colon have been described after fecal stream diversion but studies concerning the myenteric plexus after this procedure are scarce. Therefore, 28 Wistar rats were submitted to fecal diversion and followed for different periods (30, 60, 120 and 180 days), in order to study the myenteric plexus of the excluded segments. Seven non-operated rats were employed as control. The myenteric plexus was subsequently evaluated with the NADH and NADPH histochemical techniques. The colonic area of excluded segments is significantly decreased. The density of NADH-stained neurons continuously increases during the entire postoperative period but does not match the extent of surface reduction. Neuronal area measurements suggest hypertrophy of the remaining neurons in the late postoperative period. Morphological alterations of myenteric ganglia and neurons were also evident. An important surface area reduction combined with slight density increase points toward significant neuronal loss after fecal diversion. While studies correlating neuronal loss and functional changes are still lacking, surgeons should bear in mind the modifications of the myenteric plexus when performing fecal stream diversion surgeries.     

Close relationship between motor impairments and loss of functional motoneurons in a Charcot-Marie-Tooth type 1A model

Charcot-Marie-Tooth disease type 1A is the most frequent hereditary neuropathy affecting the peripheral nervous system. A partial duplication of chromosome 17 (17p11.2) involving the PMP22 gene is responsible for dysmyelination-demyelination processes leading to motor and sensory impairments.Murine models of this disease are now widely used to investigate the mechanisms occurring at the behavioural and physiological levels. In this study, adult transgenic mice (6 months old) having integrated 7 copies of the human PMP22 gene were used to compare the motor performance, evaluated by using a complex locomotor test (the rotarod test), with both the number of functional motoneurons innervating the soleus muscle and the level of myelination in the sciatic nerve. Two levels of motor deficits were detected and led us to divide the population into two subgroups.In both impaired groups, the level of motor deficit was strongly correlated with the number of functional motoneurons evaluated by retrograde labeling from the muscle, but not with the number of myelinated fibers or the thickness of the myelin sheath (g-ratio). It therefore appears that the number of motor units may be a key element in motor impairments observed in Charcot-Marie-Tooth disease type 1A disease. These findings may have implications for therapeutic procedures, which should focus on the survival of the motoneuronal pool and/or the maintenance of functional neuro-muscular connexions to reduce motor impairments in humans.     

Pregnancy after liver transplantation with tacrolimus immunosuppression: a single center's experience update at 13 years

BACKGROUND: Chronic liver disease often leads to amenorrhea in women of childbearing age. There are several reports of successful pregnancy after liver transplantation (LTx) with cyclosporine A immunosuppression. Tacrolimus has been increasingly used in solid-organ transplantation, and the effect of the drug on pregnancy is still of interest to clinicians. This study updates our single-center experience. METHODS: All pregnancies after LTx with tacrolimus immunosuppression were followed prospectively. Patients' clinical courses during pregnancy and labor along with gestational period and birth weight were catalogued. Changes in liver function, renal function, and immunosuppression also were recorded. The birth weight percentile was calculated on the basis of the gestational period using a standard chart. RESULTS: Thirty-seven mothers delivered 49 babies. Three mothers delivered three times, and six mothers delivered two times. Thirty-six mothers (97%) survived the pregnancy, and 36 allografts (97%) survived. The one death and graft loss was in a patient who demonstrated infra-aortic arterial graft, which clotted by the gravid uterus during labor. The patient developed a gangrenous liver and died before she could undergo retransplantation. The mean gestational period was 36.4+/-3.2 weeks, excluding two premature deliveries at 23 and 24 weeks gestation. Twenty-two babies (46.9%) were delivered by cesarean section, and the other babies were delivered vaginally. In addition to the two premature babies, one baby, who was born to a mother with Alagille syndrome, died from congenital birth defects. The rest of the newborns survived. The mean birth weight was 2,797+/-775 g, with 38 babies (78%) weighing more than 2,000 g. The mean birth weight percentile to gestational period was 54+/-23. Four babies (8.5%) had a birth weight percentile of less than 25, and 28 babies (59.6%) had a birth weight percentile greater than 50. Twelve patients demonstrated an increase in hepatic enzymes without jaundice during the pregnancy. All of them responded to augmentation of immunosuppression. CONCLUSION: The present report reconfirms the safety of tacrolimus during pregnancy after LTx. Preterm delivery and low birth weight seem to be a persistent problem in all solid-organ transplantation under any form of immunosuppression. However, toxemia of pregnancy and new onset of hypertension seem to be have a low occurrence with the use of tacrolimus.     

Predictors and confounders of unprotected sex: a UK web-based study.

OBJECTIVES: To evaluate the effects of gender, sex orientation, age, relationship status, age at first sex education, age at first sex, frequency of sex and number of sexual partners on the likelihood of unprotected sex in the United Kingdom (UK). METHODS: A web-based survey was conducted in the UK in 2004 with 10,138 respondents. Eight variables were chosen as potential predictors of unprotected sex. Responses from sexually active participants were combined and crude/adjusted odds ratios (AORs) were calculated for binary/categorical variables. RESULTS: No differences were found in AOR for gender, sex orientation, relationship status and frequency of sex. Differences were found in AOR for age, sex education, age at first sex and number of sexual partners, e.g. AOR 21-24 years of age versus under 16 (1.92, CI 1.38-2.68; p-value < 0.001); AOR sex education never received versus first sex education under 10 years of age (1.45, CI 1.12-1.87; p-value < 0.01); AOR first sex at 20 years versus at 14 years (0.56, CI 0.40-0.79; p-value < 0.01); and AOR 11-20 partners versus one partner (24.91, CI 20.02-30.98; p-value < 0.001). The association of sex orientation and relationship status with unprotected sex is confounded by the number of sexual partners. CONCLUSIONS: UK health and education authorities should develop strategies to provide sex education starting in primary school and continuing throughout secondary school. Prevention programs and policies should focus on particular sexual behaviours. Furthermore, an understanding of sexual diversity and reducing stigmatisation of sexual orientation is necessary.     

Impact of the single-dose universal mass vaccination strategy against hepatitis A in Brazil

Universal vaccination of children against hepatitis A was introduced in 2014 in Brazil as a single-dose schedule. We analyzed the numbers of reported cases of hepatitis A virus infection (HAV) from 2010 to 2017 to evaluate the initial impact of that intervention. Data were assessed and has been freely available on the Brazilian Ministry of Health website. The HAV incidence was steady around 6000 cases per year until 2014. Between 2014 and 2016, there was a 85.5% cumulative drop, independent of gender and geographical macroregions. The fall was especially significant among toddlers (96.8%). In 2017, cases increased due to an outbreak among male adults in São Paulo. Decrease in incidence continued to occur for females and for those under 15?years of age. Data show that there was a significant decrease in HAV cases number in Brazil from 2015 after the introduction of single-dose HAV vaccine program.     

Plasmodium vivax circumsporozoite variants and Duffy blood group genotypes in the Brazilian Amazon region

The circumsporozoite protein (CSP) of the Plasmodium vivax infective sporozoite is considered to be a major target for the development of recombinant malaria vaccines. The Duffy blood group molecule acts as the red blood cell receptor for P. vivax. We review the frequency of P. vivax CSP variants and report their association with the Duffy blood group genotypes from Brazilian Amazon patients carrying P. vivax malaria. Peripheral blood samples were collected from 155 P. vivax-infected individuals from five Brazilian malaria-endemic areas. The P. vivax CSP variants and the Duffy blood group genotypes were assessed using PCR/RFLP. In single infections, the VK210 variant was the commonest followed by the P. vivax-like variant. The typing of P. vivax indicated that the frequency of variants among the study areas was significantly different from one to another. This is the first detection of the VK247 and P. vivax-like variant in single infections in endemic areas of Brazil. Association of the CSP P. vivax variants with the heterozygous Duffy blood group system genotype was significant for VK210 single infection. These observations provide additional data on the Plasmodium-host interactions concerning the Duffy blood group and P. vivax capability of causing human malaria.     

Impact of prior chronic statin therapy and high-intensity statin therapy at discharge on circulating endothelial progenitor cell levels in patients with acute myocardial infarction: a prospective observational study

Background

Endothelial progenitor stem cells (EPCs) are mobilized to the peripheral circulation in response to myocardial ischemia, playing a crucial role in vascular repair. Statins have been shown to stimulate EPCs. However, neither the impact of previous statin therapy on EPC response of acute myocardial infarction (AMI) patients nor the effect of post-AMI high-intensity statin therapy on the evolution of circulating EPC levels has yet been addressed. Therefore, we aimed to compare circulating EPC levels between patients receiving long-term statin therapy before the AMI and statin-naive patients and to assess the impact of high-intensity statin therapy at discharge on the evolution of circulating EPCs post-AMI.

Methods

This is a prospective observational study of 100 AMI patients. Circulating EPCs (CD45dimCD34?+?KDR?+?cells) and their subpopulation coexpressing the homing marker CXCR4 were quantified by the high-performance flow cytometer FACSCanto II in whole blood, in two different moments: within the first 24 h of admission and 3 months post-AMI. Patients were followed up clinically for 2 years.

Results

Patients previously treated with statins had significantly higher levels of EPCs coexpressing CXCR4 (1.9?±?1.4 vs. 1.3?±?1.0 cells/1,000,000 events, p?=?0.031) than statin-naive patients. In addition, the subanalysis of diabetics (N?=?38) also revealed that patients previously on statins had significantly greater numbers of both CD45dimCD34?+?KDR?+?CXCR4+ cells (p?=?0.024) and CD45dimCD34?+?KDR?+?CD133+ cells (p?=?0.022) than statin-naive patients. Regarding the evolution of EPC levels after the AMI, patients not on a high-intensity statin therapy at discharge had a significant reduction of CD45dimCD34?+?KDR?+?and CD45dimCD34?+?KDR?+?CXCR4+ cells from baseline to 3 months follow-up (p?=?0.031 and p?=?0.005, respectively). However, patients discharged on a high-intensity statin therapy maintained circulating levels of all EPC populations, presenting at 3 months of follow-up significantly higher EPC levels than patients not on an intensive statin therapy. Moreover, the high-intensity statin treatment group had significantly better clinical outcomes during the 2-year follow-up period than patients not discharged on a high-intensity statin therapy.

Conclusion

Chronic statin therapy prior to an AMI strongly enhances the response of EPCs to myocardial ischemia, even in diabetic patients. Furthermore, high-intensity statin therapy after an AMI prevents the expected decrease of circulating EPC levels during follow-up. These results reinforce the importance of an early and intensive statin therapy in AMI patients.