Improvement in lipids after switch to boosted atazanavir or darunavir in children/adolescents with perinatally acquired HIV on older protease inhibitors: results from the Pediatric HIV/AIDS Cohort Study.

OBJECTIVES: Dyslipidaemia is common in perinatally HIV-infected (PHIV) youth receiving protease inhibitors (PIs). Few studies have evaluated longitudinal lipid changes in PHIV youth after switch to newer PIs. METHODS: We compared longitudinal changes in fasting lipids [total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and TC:HDL-C ratio] in PHIV youth enrolled in the Pediatric HIV/AIDS Cohort Study (PHACS) Adolescent Master Protocol (AMP) study who switched to atazanavir/ritonavir (ATV/r)- or darunavir/ritonavir (DRV/r)-based antiretroviral therapy (ART) from an older PI-based ART and those remaining on an older PI. Generalized estimating equation models were fitted to assess the association of a switch to ATV/r- or DRV/r-based ART with the rate of change in lipids, adjusted for potential confounders. RESULTS: From 2007 to 2014, 47 PHIV children/adolescents switched to ATV/r or DRV/r, while 120 remained on an older PI [primarily lopinavir/r (72%) and nelfinavir (24%)]. Baseline age ranged from 7 to 21 years. After adjustment for age, Tanner stage, race/ethnicity, and HIV RNA level, a switch to ATV/r or DRV/r was associated with a more rapid annual rate of decline in the ratio of TC:HDL-C. (ß = -0.12; P = 0.039) than remaining on an older PI. On average, TC declined by 4.57 mg/dL/year (P = 0.057) more in the switch group. A switch to ATV/r or DRV/r was not associated with the rate of HDL-C, LDL-C, or TG change. CONCLUSIONS: A switch to ATV/r or DRV/r may result in more rapid reduction in TC and the TC:HDL-C ratio in PHIV youth, potentially impacting long-term cardiovascular disease risk.

Testis peritubular myoid cells increase their motility and express matrix-metalloproteinase 9 (MMP-9) after interaction with embryonal carcinoma cells.

Today cancer research studies have highlighted the role of the cancer-stroma interaction in the regulation of invasive processes. However, very little is known about cell-to-cell relationships between germinal cancer cells and the somatic ones belong to their close environment, particularly at early invasion stages. Here, we have studied the potential role of the seminiferous peritubular myoid cells (PTCs), as potential part of the reactive stroma, like tumor myofibroblast, in the progression of embryonal carcinoma (EC). To this end, we show results on the in vitro interactions between F9 murine embryonal carcinoma cells (EC cells) and primary cultures of murine PTCs, using contact-dependent and contact-independent 2D co-cultures. In these circumstances, when EC cells interact with PTCs they change their migratory behavior and matrix-metalloproteinase 9 (MMP-9) was up-regulated in PTCs. Additionally, among a variety of cytokines implicated in tumor-stroma cross-talk, we have examined in more detail the influence of tumor necrosis factor alpha (TNF-α). In this regard, it was observed that this cytokine induced a MMP-9 secretion by PTCs in a pattern dependent on its concentration, whereas does not increase the migration capacity of cancer cells. All together, our results provide evidence for a role played by peritubular myoid cells and cancer-cell secreted TNF- α for a change in the tumor microenvironment during the early stages of EC progression.

Angiogenesis and vascular network of teratocarcinoma from embryonic stem cell transplant into seminiferous tubules.

BACKGROUND: Carcinoma in situ (CIS) of the testis is considered to be a precancerous germinal cell lesion, but the precise cellular and molecular mechanisms underlying transformation of CIS into invasive pluripotent cancer cells remain to be elucidated. Moreover, a satisfactory animal model for the experimental study of germinal tumours has not been developed to date. METHODS: We have developed a tumour model that involves the microinjection of green fluorescent protein-labelled embryonic stem (ES) cells (which are functionally equivalent to CIS cells) into syngenic mouse seminiferous tubules, a unique cell microenvironment in which germinal cells mature and CIS arise. To characterise the vascularisation of teratocarcinomas, which arise after cell transplant, we used immunohistochemistry, together with a qualitative and quantitative analysis of scanning electron microscopy images of corrosion casting samples. RESULTS: Embryonic stem cells transplanted into seminiferous tubules did not differentiate into germinal cells, but rather they behaved as invasive embryonal carcinoma (EC) stem cells. The vascular pattern of the experimental teratocarcinomas showed a highly disorganised architecture, and some of the neoplastic capillaries were derived, at least in part, from the original transplanted ES cells. CONCLUSION: The transplantation of pluripotent ES cells into seminiferous tubules efficiently recapitulates the early stages of development of teratocarcinomas. Consequently, this method constitutes a novel in vivo model to study the mechanisms of invasion and progression of experimental germinal tumours.

Prevalence of community-associated methicillin-resistant Staphylococcus aureus colonization outside the healthcare environment.

Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections are increasingly recognized in persons without established risk factors. Population-based prevalence studies of CA-MRSA colonization in persons without risk factors are relatively limited. Subjects aged 2-65 years were enrolled from a student recreation centre, public office building, and out-patient clinics. Persons or close contacts with a history of hospitalization, nursing-home residence, surgery, emergency-department visit, or healthcare employment during the previous year and persons with chronic debilitating illness, indwelling catheter, or surgical device were excluded. Swabs of anterior nares were obtained. Demographic and clinical information was collected. During January-June 2005, three (1.2%) of 259 subjects were colonized with MRSA. All three subjects were adults enrolled at the recreation centre. Healthy persons living in households without recent exposure to healthcare environments were at low risk for MRSA colonization. Studies from other geographic locations are needed to elucidate differences in prevalence of CA-MRSA.

Risk factors for preterm birth, low birth weight, and intrauterine growth retardation in infants born to HIV-infected pregnant women receiving zidovudine. Pediatric AIDS Clinical Trials Group 185 Team.

OBJECTIVE: To evaluate independent contributions of maternal factors to adverse pregnancy outcomes (APO) in HIV-infected women receiving antiretroviral therapy (ART). DESIGN: Risk factors for preterm birth (< 37 weeks gestation), low birth weight (LBW) (< 2500 g), and intrauterine growth retardation (IUGR) (birth weight < 10th percentile for gestational age) examined in 497 HIV-infected pregnant women enrolled in PACTG 185, a perinatal clinical trial. METHODS: HIV RNA copy number, culture titer, and CD4 lymphocyte counts were measured during pregnancy. Information collected included antenatal use of cigarettes, alcohol, illicit drugs; ART; obstetric history and complications. RESULTS: Eighty-six percent were minority race/ethnicity; 86% received antenatal monotherapy, predominantly zidovudine (ZDV), and 14% received combination antiretrovirals. Preterm birth occurred in 17%, LBW in 13%, IUGR in 6%. Risk of preterm birth was independently associated with prior preterm birth [odds ratio (OR) 3.34; P < 0.001], multiple gestation (OR, 6.02; P = 0.011), antenatal alcohol use (OR, 1.91; P = 0.038), and antenatal diagnosis of genital herpes (OR, 0.24; P = 0.022) or pre-eclampsia (OR, 6.36; P = 0.025). LBW was associated with antenatal diagnosis of genital herpes (OR, 0.08; P = 0.014) and pre-eclampsia (OR, 5.25; P = 0.049), and baseline HIV culture titer (OR, 1.41; P = 0.037). IUGR was associated with multiple gestation (OR, 8.20; P = 0.010), antenatal cigarette use (OR, 3.60; P = 0.008), and pre-eclampsia (OR, 12.90; P = 0.007). Maternal immune status and HIV RNA copy number were not associated with APO. CONCLUSIONS: Risk factors for APO in antiretroviral treated HIV-infected women are similar to those reported for uninfected women. These data suggest that provision of prenatal care and ART may reduce APO.

Antiviral therapy in a child with pediatric human immunodeficiency virus (HIV): case study of audiologic findings.

Over the past decade, much research has been conducted to determine the auditory consequences of human immunodeficiency virus (HIV) infection and acquired immune deficiency syndrome (AIDS). This research, primarily using adult patients, has focused on the involvement of the central auditory nervous system (CANS). Measures of auditory evoked potentials, particularly the auditory brainstem response (ABR), can document changes in the CANS as the disease progresses and during treatment with antiviral therapies such as zidovudine (AZT) and didanosine (ddI). This case study presents the audiologic findings for a child with HIV infection. Evaluations were performed over a 2-year period prior to the initiation of antiviral therapy and following treatment. Audiologic measures included behavioral audiometry, tympanometry, otoacoustic emissions, and ABR latency/intensity functions and rate studies. Findings indicated a gradual shortening of all ABR component latencies following the initiation of antiviral therapy. In addition, a high-frequency hearing loss was detected during the final evaluation subsequent to 19 months of treatment with AZT and ddI.

Effects of imipramine administration on mu-opioid receptor immunostaining in the rat forebrain.

Imipramine hydrochloride (CAS 113-52-0) is a widely used antidepressant and can interact with opioid receptors. However, complete information concerning possible regional alterations in mu-opioid receptor expression in the rat forebrain after chronic treatment with this substance is still lacking. This being the case, analysis of mu-opioid receptor immunostaining in several regions of the rat brain after imipramine administration in vivo was made, and an increase in the density of cells expressing mu-opioid receptors in the caudatus-putamen, the dentate gyrus and the frontal, parietal and piriform cortices after chronic imipramine treatment, with respect to controls, was found. These data suggest that mu-opioid receptor expression in the rat forebrain is altered by in vivo chronic imipramine treatment.

Toluene alters brainstem enkephalinergic system in rats.

Acute exposure to high doses of toluene can generate respiratory depression. However, neurotoxic mechanism of its action in the brainstem is not completely clear. In this work, acute, but not subchronic, exposure of rats to toluene increased leu-enkephalin immunostaining in several myelencephalic nuclei implicated in cardiorespiratory control. Due to the physiological role of enkephalins in the central regulation of breathing, it is suggested that the enkephalinergic system could play a role in neurotoxic respiratory depression induced by high dose acute toluene exposure.

Effects of acute lidocaine administration on the rat prosencephalic enkephalinergic system.

Lidocaine is a local anesthetic widely used in therapeutics and as antiarrhythmic agent. However, information concerning possible alterations in the enkephalinergic system after acute treatment with this substance is not complete. This being the case, we focused on analyzing enkephalin immunostaining in several regions of the rat brain after lidocaine administration. We could not find significative changes in rat prosencephalon. These data might suggest that the enkephalinergic system is not altered by lidocaine.

Correlations between intestinal parasitosis, physical growth, and psychomotor development among infants and children from rural Nicaragua.

The correlations between malnutrition, parasitosis (especially helminth infections), and child development are complex, and studies of these interrelationships will allow health agencies to maximize screening and intervention strategies for developing countries. We examined these correlations in a cross-sectional program in Carazo State, Nicaragua. Nine hundred sixty-one children in two age strata (ages 0-24 months and ages 2-10 years) from one urban and three rural communities were screened for intestinal parasites (direct smear and ZnSO4 flotation), malnutrition, and developmental delays. Nutritional status was determined as weight-for-age (WFA), weight-for-height (WFH), and height-for-age (HFA). Developmental status (normal, suspect) was determined for the four subtests of the Denver II Screening Test. The prevalence of malnutrition was 14.6% (WFA), 8.4% (WFH), and 36.3% (HFA). Parasitosis was more prevalent in children less than 24 months of age with low HFA, whereas in older children low WFA was more closely associated with parasitic infections. Ascaris and Trichuris were more prevalent in malnourished children. On the Denver II, suspect test results in all four categories (language, social, gross motor, and fine motor) were associated with low WFA, and suspect language tests were associated with both intestinal parasites (P = 0.0003) and Ascaris infection in particular (P = 0.044). Developmental disabilities are a significant and frequently undetected health problem in developing countries, and malnutrition associated with intestinal helminth infections may be an important contributory factor for these disabilities.

PIP: Studies of the correlations between malnutrition, parasitosis, and child development will help health agencies to maximize screening and intervention strategies for developing countries. Such correlations were examined in a cross-sectional study in Carazo State, Nicaragua. 961 children aged 0-24 months and 2-10 years from 1 urban and 3 rural communities were screened for infection with intestinal parasites, malnutrition, and developmental delays. Developmental status was determined for the 4 subtests of the Denver II Screening Test. The prevalence of malnutrition was 14.6% according to weight-for-age (WFA), 8.4% for weight-for-height (WFH), and 36.3% for height-for-age (HFA). Parasitosis was more prevalent among children under age 2 years with low HFA, while low WFA was more closely associated with parasitic infections in older children. Ascaris and Trichuris were more prevalent in malnourished children. On the Denver II, suspect test results in language, social, gross motor, and fine motor were associated with low WFA, while suspect language tests were associated with both intestinal parasites, and Ascaris infection in particular. Developmental disabilities are a significant and often undetected health problem in developing countries. Malnutrition associated with intestinal helminth infections may be an important contributory factor for such disabilities.

Soluble and membrane-bound pyroglutamyl-peptidase I activity in the developing cerebellum and brain cortex.

Developmental changes of soluble and particulate pyroglutamyl (pGlu)-peptidase I activities in the rat brain cortex and in the cerebellum are described in this work. The enzyme activity has been measured spectrofluorimetrically using pGlu-b-naphthylamide as substrate (in the presence and absence of EDTA and DTT, necessary activators of the enzyme) in both soluble and particulate fractions. In the soluble fraction of the cerebellum and brain cortex, pGlu-peptidase I activity is high in the perinatal period and decreases two or three folds subsequently, at a later stage in the cerebellum than in the brain cortex, reaching adult levels at the end of the first postnatal month. The decrease in the activity of pGlu-peptidase observed in this work coincides with increasing levels of brain thyroliberin concentration after the second postnatal week. The particulate pGlu-peptidase I activity, obtained after osmotic shock and high-salt treatment, shows less significant changes during brain development in the areas under study. It is suggested that cytosolic pGlu-peptidase I could play a part in the normal development of the rat central nervous system.

Membrane-bound tyrosine aminopeptidase activities in the rat brain throughout the estrous cycle.

Puromycin sensitive and insensitive membrane-bound aminopeptidase activity levels during the estrous cycle in several brain areas have been described in this research. We have found the highest aminopeptidase M activity levels during the proetrous stage in the hypothalamus, the amygdala and the pituitary gland. Since this enzyme has been involved in opioid peptide metabolism, it is suggested that aminopeptidase M could play a part in the decrease in the inhibitory influence of the endogenous opioids peptides that participate in the LH surge.

Membrane-bound pyroglutamyl-arylamidase activity during the first postnatal month in several rat brain areas.

Membrane-bound pyroglutamyl-aminopeptidase activity cleaves the pyoglutamate amino acid bond of thyroliberin (TRH). Information concerning developmental variations in TRH has been reported. However, little is known about the ontogeny of the membrane-bound enzyme activities capable of hydrolyzing the mentioned tripeptide. In this work we have described decreases in membrane-bound pyroglutamyl-aminopeptidase (arylamidase) activity, from day 9 to day 20 after birth, in the hypothalamus, the striatum, the frontal, occipital and parieto-temporal cortices and the pituitary gland of the male and the female rat. The developmental profile is similar in rats of both sexes. We have not found significative changes between 20 and 25 postnatal days. The observed decreasing activity is developmentally coincident with the increases in thyroliberin and decreases in Hys-Pro diketopiperazine concentration in different brain areas. It is suggested that membrane-bound pyroglutamyl-peptidase activity could play a part in the normal development of thyroliberin physiology.

Mass and molecular weight of isolated nuclear rings.

Nuclear rings are cell structures found at the nuclear cortex wedged between the nuclear envelope and the chromatin fiber network. In previous publications we have dealt with their morphology, relationships with the nuclear membranes, chromatin fibers and cytoskeletal filaments; and more recently, with their measurements at high electron microscope resolution. In this article we have calculated the mass and molecular weight of 336 isolated nuclear rings from human circulating lymphocytes using a photometric procedure and polystyrene latex spheres as the standard for weight calibration. Our results show a range of mass of 0.4-35.5 x 10(-16) g (equivalent to 0.2-21.2 x 10(8) Da with a positively skewed distribution (median: 3.3 x 10(-16) g or 2.0 x 10(8) Da). Mass and volume of nuclear rings were highly correlated. In addition, it was possible to calculate the area, the whole mass and the mass per unit area of the nuclear envelope present in the center of the nuclear rings. The mass of this area also shows a lognormal distribution (median of mass/unit area: 37.3 x 10(-8) pg/nm2 or 1.9 x 10(5) Da/nm2). We discuss the significance of this results as parameters for the characterization of the nuclear rings and their possible implications for a new interpretation of nuclear cortex architecture, nucleocytoplasmic traffic and macromolecule segregation between the two main cell compartments.

Regulation of inositol phosphate levels by prostaglandins in cultured endometrial cells.

Stimulation of cultured rabbit endometrial cells by one of the rabbit endometrial cell culture proliferation factors, prostaglandin F2 alpha (PGF2 alpha), resulted in a very rapid increase in the intracellular levels of [3H]-inositol triphosphate (IP3), [3H]-inositol biphosphate (IP2), and [3H]-inositol monophosphate (IP1) in cells prelabeled with [3H]-inositol. These increases in inositol phosphate levels were detected in periods of stimulation as short as 30 seconds, reached a maximum by 1 1/2-2 min and declined to control levels by 6-10 min. The stimulation was dose-dependent with maximal increases observed near 10(-6) M PGF2 alpha. The cholinergic agent, carbachol, also led to time and dose-independent increases in IP3. Lithium, cadmium, silver, copper, and zinc ions had no effect either on the breakdown of IP3 or on the accumulation of IP1. In contrast, vanadate at 10(-6) or 10(-5) M did lead to a decrease in the breakdown of IP1 and a concomitant increase in IP1, IP2, and IP3. PGF2 alpha was found previously to induce an increase in rabbit endometrial cell DNA synthesis which was inhibited by concomitant or prior addition of prostaglandin E1 (PGE1). PGE1, in a dose-dependent manner, was found to inhibit the observed IP3 increase by PGF2 alpha at 1 1/2 min of stimulation. PGF2 alpha treated and control cultures did not differ in cAMP or cGMP levels, cellular 45Ca uptake, nor cellular 22Na uptake. We propose that IP3 may be one of the intracellular messenger(s) synthesized following the treatment of rabbit endometrial cell cultures with the proliferation agent PGF2 alpha and that it may play a crucial role with cAMP in growth regulation.