Activation of blood T lymphocytes down-regulates CXCR4 expression and interferes with propagation of X4 HIV strains

The chemokine receptor CXCR4 serves as a coreceptor for HIV-1 entry into CD4⁺ cells, in particular for strains emerging late in the infection. Cell surface expression of CXCR4 has, therefore, important implications for HIV-1 pathogenesis. Using blood lymphocytes cultured under various conditions, we studied the expression and regulation of CXCR4. Flow cytometry showed that only about 20 % of freshly isolated lymphocytes expressed CXCR4 on the cell surface whereas in 80 % of resting blood lymphocytes CXCR4 was located intracellularly. Within a few hours in culture, the intracellular CXCR4 was translocated to the surface and was expressed in the large majority of both naive and memory lymphocytes. A decrease in surface expression of CXCR4 was found when lymphocytes cultured overnight for maximal receptor expression were stimulated with phytohemagglutinin, anti-CD3 antibodies, phorbol 12-myristate 13-acetate and stromal cell-derived factor-1. The superantigen staphylococcal enterotoxin A, a more selective stimulus, induced a marked decrease in CXCR4 expression preferentially in cells positive for the CD25 activation marker. Confocal laser scanning microscopy demonstrated the presence of CXCR4 in the cytosol and on the surface of resting lymphocytes and also showed CXCR4 redistribution after activation. The number of cells infected by the X4 HIV strain NL4.3 paralleled the expression of CXCR4 in CD4⁺ T lymphocytes. Sustained reduction of CXCR4 cell surface expression upon activation with phytohemagglutinin correlated with a low number of CD4⁺ T lymphocytes expressing HIV p24 gag antigen. Our results indicate that activation of CD4⁺ T lymphocytes reduces surface expression of CXCR4 in part by receptor internalization and that cell activation-dependent CXCR4 down-regulation limits spread of infection by X4 viruses.     

Impact of Repeated Acute Exposures to Low and Moderate Exercise-Induced Hypohydration on Physiological and Subjective Responses and Endurance Performance

This study aimed to examine whether repeated exposures to low (2%) and moderate (4%) exercise-induced hypohydration may reverse the potentially deleterious effect of hypohydration on endurance performance. Using a randomized crossover protocol, ten volunteers (23 years, V˙O_2max: 54 mL∙kg⁻¹∙min⁻¹) completed two 4-week training blocks interspersed by a 5-week washout period. During one block, participants replaced all fluid losses (EUH) while in the other they were fluid restricted (DEH). Participants completed three exercise sessions per week (walking/running, 55% V˙O_2max, 40 °C): (1) 1 h while fluid restricted or drinking ad libitum, (2) until 2 and (3) 4% of body mass has been lost or replaced. During the first and the fourth week of each training block, participants completed a 12 min time-trial immediately after 2% and 4% body mass loss has been reached. Exercise duration and distance completed (14.1 ± 2.7 vs. 6.9 ± 1.5 km) during the fixed-intensity exercise bouts were greater in the 4 compared to the 2% condition (p < 0.01) with no difference between DEH and EUH. During the first week, heart rate, rectal temperature and perceived exertion were higher (p < 0.05) with DEH than EUH, and training did not change these outcomes. Exercise-induced hypohydration of 2% and 4% body mass impaired time-trial performance in a practical manner both at the start and end of the training block. In conclusion, exercise-induced hypohydration of 2% and 4% body mass impairs 12 min walking/running time-trial, and repeated exposures to these hypohydration levels cannot reverse the impairment in performance.     

Immunohistochemical analysis of CD4+ and CD8+ T-cell subsets in high risk human papillomavirus-associated pre-malignant and malignant lesions of the uterine cervix

OBJECTIVES: Humoral and cellular immune responses are likely to play a key role for the clearance or persistence and progression of high risk (HR) HPV-associated cervical lesions. Although there are many studies describing the systemic T-cell responses to HPV16 and 18 proteins, few data are available regarding the cellular mucosal immune responses. We used immunohistochemistry to characterize populations of T-immune cells (CD4+, CD8+, CD45RO+) in HR-HPV-infected precancerous and cancerous lesions of the uterine cervix. METHODS: Four biopsies from normal cervix, 9 CIN1 which have regressed (rCIN), 5 CIN1 which have progressed (pCIN) to high grade lesions, 13 CIN3 and 11 invasive carcinomas were included. All dysplasias and carcinomas were HR-HPV-positive and low-risk-HPV-negative. They were stained with monoclonal antibodies specific for CD4, CD8 and CD45RO and examined by microscopy. STATISTICAL ANALYSIS: The Kruskal-Wallis test and the Siegel's and Castelan's method were used. RESULTS.: CD4+ cells predominated in regressing CIN1 both within the stroma and the epithelium with the highest CD4+/CD8+ ratio compared with pCIN1, CIN3 and invasive carcinoma. At the exception of CD45RO+ cells, T cells were detected with similar frequencies in both pCIN1 and CIN3. However, in 7 out of 10 CIN3, CD4+ and CD8+ cells were visible as organized lymphoid follicle structure. The CD8+ and CD45RO+ cells far exceeded the CD4+ cells in invasive cancers. CONCLUSIONS: Density and distribution of immune T cells depend on the malignant potential of HR-HPV lesions. These results suggest that the studied lymphocyte subsets have an important role to fulfil during the natural history of HR-HPV-associated lesions.     

Fatal Cowpox Virus Infection in Human Fetus,France, 2017

Cowpox virus (CPXV) has an animal reservoir and is typically transmitted to humans by contact with infected animals. In 2017, CPXV infection of a pregnant woman in France led to the death of her fetus. Fetal death after maternal orthopoxvirus (smallpox) vaccination has been reported; however, this patient had not been vaccinated. Investigation of the patient’s domestic animals failed to demonstrate prevalence of CPXV infection among them. The patient’s diagnosis was confirmed by identifying CPXV DNA in all fetal and maternal biopsy samples and infectious CPXV in biopsy but not plasma samples. This case of fetal death highlights the risk for complications of orthopoxvirus infection during pregnancy. Among orthopoxviruses, fetal infection has been reported for variola virus and vaccinia virus; our findings suggest that CPXV poses the same threats for infection complications as vaccinia virus.     

Prenatal diagnosis of CHARGE syndrome by identification of a novel CHD7 mutation in a previously unaffected family

CHARGE syndrome comprises ocular coloboma (C), heart malformation (H), choanal atresia (A), retardation of growth and/or anomalies of the central nervous system (R), genital anomalies (G) and ear anomalies (E). Prenatal diagnosis of CHARGE syndrome may be suspected in the presence of specific major anomalies at ultrasound examination. We describe prenatal diagnosis of CHARGE syndrome confirmed by identification of a mutation in CHD7 gene in a previously unaffected family.     

Construction à Étampes d’un premier programme d’éducation thérapeutique du patient à destination des personnes soignées pour une schizophrénie et de leurs aidants

In 1997, an expert group from WHO Europe made a report and international recommendations for training and practice in therapeutic education of the patient. The French version of the text was published the following year. In France, it is the law of 21 July 2009, called “Patient Health Territory Hospital”, which formalizes this practice and inscribes it in the course of care of patients suffering from chronic diseases. The TPE is regulated by official texts and must respect the recommendations of the High health authority. That is the French peculiarity. Thus, any TPE program must be authorized by the Regional Health Agency (ARS) on which the establishment is dependent. Practices that would not be authorized by these ARS are not entitled to be called TPE, even if they meet all of the criteria. Although included in the law in 2009, this practice of TPE remains long unknown in France to most hospital practitioners. Thus, concerning our psychiatric hospital, located 50 km south of Paris, it is a certification visit that requires us in 2011 to register the TPE in our program of establishment. A small group of professionals of varied training (psychiatrist, addictologist, general practitioner, pharmacist, psychologist, nurse, social worker, educator, legal agent, dietician, caregiver) is working on this task in the course of the year 2012. We decide to build a program for people suffering from schizophrenia. We form a partnership with an association of Patient families, the UNAFAM. We choose to do only group support, more effective in the pathology we have retained. When the practitioners involved present their project in front of their peers, they have not yet received the mandatory 40 hours training required by the HAS for any therapeutic education workshop facilitator. This may explain that our project is being received with so little enthusiasm. We are faced with many reproaches. However, we are pursuing the construction of the workshops, which will be disturbed by the compulsory training of 40 hours that only be in 2013. We realize that the support of our hierarchy is essentially moral, since no dedicated time will be assigned to this program, which we have to carry out with the same means. We start our first workshops in September 2014, with ARS funding as an experimental project. Our program includes the following eight workshops: “Living with your illness”, “understanding and managing your treatment”, “social skills”, “lifestyle”, “addicts”, “Libido”, “do”, “do with”, “do together”, “caregivers”. The nurse coordinator carries out the educational diagnosis and the participant decides which workshops he wants to participate in. This nurse coordinator participates in all the workshops in addition to the specific animators. The workshops follow each other and two programs are closed over the year. There is no mandatory order and participants can integrate a program at any time. Often, they ask to participate in other workshops than those initially chosen after having experienced the group. The approach of the workshop animators is based on the motivational approach. After four years of operation, the effectiveness of this program is being evaluated. The feedback from participants is in any case very favorable, and they all underline their pleasure in having interlocutors who are no longer in the usual caregiving posture. But we also measure the fragility of this program, depending on a very small number of people.     

Bioluminescence calcium imaging of network dynamics and their cholinergic modulation in slices of cerebral cortex from male rats

The activity of neuronal ensembles was monitored in neocortical slices from male rats using wide-field bioluminescence imaging of a calcium sensor formed with the fusion of green fluorescent protein and aequorin (GA) and expressed through viral transfer. GA expression was restricted to pyramidal neurons and did not conspicuously alter neuronal morphology or neocortical cytoarchitecture. Removal of extracellular magnesium or addition of GABA receptor antagonists triggered epileptiform flashes of variable amplitude and spatial extent, indicating that the excitatory and inhibitory networks were functionally preserved in GA-expressing slices. We found that agonists of muscarinic acetylcholine receptors largely increased the peak bioluminescence response to local electrical stimulation in layer I or white matter, and gave rise to a slowly decaying response persisting for tens of seconds. The peak increase involved layers II/III and V and did not result in marked alteration of response spatial properties. The persistent response involved essentially layer V and followed the time course of the muscarinic afterdischarge depolarizing plateau in layer V pyramidal cells. This plateau potential triggered spike firing in layer V, but not layer II/III pyramidal cells, and was accompanied by recurrent synaptic excitation in layer V. Our results indicate that wide-field imaging of GA bioluminescence is well suited to monitor local and global network activity patterns, involving different mechanisms of intracellular calcium increase, and occurring on various timescales.     

Effect of using frozen–thawed bovine semen contaminated with lumpy skin disease virus on in vitro embryo production

Lumpy skin disease (LSD) is an important transboundary animal disease of cattle with significant economic impact because of the implications for international trade in live animals and animal products. LSD is caused by a Capripoxvirus, LSD virus (LSDV), and results in extensive hide and udder damage, fever and pneumonia. LSDV can be shed in semen of infected bulls for prolonged periods and transmitted venereally to cows at high doses. This study examined the effects of LSDV in frozen‐thawed semen on in vitro embryo production parameters, including viral status of media and resulting embryos. Bovine oocytes were harvested from abattoir‐collected ovaries and split into three experimental groups. After maturation, the oocytes were fertilized in vitro with frozen‐thawed semen spiked with a high (HD) or a lower (LD) dose of LSDV, or with LSDV‐free semen (control). Following day 7 and day 8 blastocyst evaluation, PCR and virus isolation were performed on all embryonic structures. After completing sufficient replicates to reach 1,000 inseminated oocytes, further in vitro fertilization (IVF) runs were performed to provide material for electron microscopy (EM) and embryo washing procedures. Overall, in vitro embryo yield was significantly reduced by the presence of LSDV in frozen‐thawed semen, irrespective of viral dose. When semen with a lower viral dose was used, significantly lower oocyte cleavage rates were observed. LSDV could be detected in fertilization media and all embryo structures, when higher doses of LSDV were present in the frozen‐thawed semen used for IVF. Electron microscopy demonstrated LSDV virions inside blastocysts. Following the International Embryo Transfer Society washing procedure resulted in embryos free of viral DNA; however, this may be attributable to a sampling dilution effect and should be interpreted with caution. Further research is required to better quantify the risk of LSDV transmission via assisted reproductive procedures.