In vivo regulation of extracellular signal-regulated protein kinase (ERK) and protein kinase B (Akt) phosphorylation by acute and chronic morphine

In vitro evidence suggests that extracellular signal-regulated protein kinases (ERKs) and Akt (also referred to as protein kinase B) are among the myriad of intracellular signaling molecules regulated by opioid receptors. The present study examined the regulation of ERK and Akt activation in the nucleus accumbens and caudate putamen following acute and chronic morphine administration in the rat. ERK and Akt are activated by phosphorylation, hence the levels of phosphorylated ERK (pERK) and Akt (pAkt) as well as total levels of ERK and Akt protein were measured by Western blot analysis. Male Sprague-Dawley rats received either a single injection of morphine or twice daily injections of morphine for 6 or 10 days. Following acute morphine, pERK levels were significantly decreased in the nucleus accumbens but not in the caudate putamen. Phosphorylated Akt levels in the nucleus accumbens were significantly increased after a single morphine injection. Naltrexone pretreatment prevented both the morphine-induced pERK down-regulation and pAkt up-regulation. Although reductions in pERK levels were evident after 6 days of morphine administration, no differences were observed in pERK levels after 10 days. In contrast to the up-regulation seen after acute morphine, pAkt levels in the nucleus accumbens were significantly decreased after chronic morphine administration. Thus, the differential activation patterns of both ERK and Akt after acute and chronic morphine administration could have important implications for understanding additional pathways mediating opioid signaling in vivo.     

Effects of bacterial superantigens on behavior of mice in the elevated plus maze and light-dark box

Bacterial superantigens, such as staphylococcal enteroxins A and B (SEA/SEB) stimulate T cells to produce high levels of cytokines in blood. Previously it had been shown that these toxins were capable of stimulating increased neuroendocrine activity and enhanced behavioral reactivity to novel gustatory and non-gustatory stimuli. Therefore, it was suggested that these superantigens may promote anxiety-like behavior. In the current set of experiments, BALB/cByJ and C57BL/6J male mice were challenged with either SEB (50 microg) or SEA (5 or 10 microg) and tested for behavior in the elevated plus maze (EPM). Results suggested an absence of increased anxiety-like behavior, with exploration of the open arms being enhanced by SEA or SEB treatment. In another test of anxiety, the light-dark box, SEB challenge of BALB/cByJ mice 90 min prior to testing, did not alter exit latency, activity nor time spent in the dark. However, in a second experiment, it was found that if animals were first tested for consumption, followed by testing in the light-dark box, SEB challenged animals displayed increased exit latency and reduced exploration. These studies suggest that in standard tests of rodent anxiety-like behavior, evidence for the induction of anxiety-like processes subsequent to challenge with SEA or SEB is not patently discernable. However, neurobiological events induced by immunological challenge might synergize with reactivity to psychogenic and/or gustatory stimuli, thereby resulting in increased anxiety-like behavior that could be unmasked by standard behavioral tests such as the light-dark box or EPM.     

Pharmacokinetics of acyclovir in rabbit skin after i.v.-bolus, ointment, and iontophoretic administrations

The aim of this study was to characterize and compare the pharmacokinetics of acyclovir (ACV) in skin and plasma after iontophoresis, i.v.-bolus, and ointment administrations in rabbit. On five occasions, each separated by at least 1-week washout, rabbits received a 10 mg/kg dose of ACV as i.v.-bolus, ACV iontophoresis for 1 h at different current densities (100, 200, 300 microA/cm2) or a commercially available ointment for two hours. Blood samples were collected serially up to 6 h. Skin ACV concentrations were monitored via microdialysis using linear microdialysis probes (1 cm window). Cathodic iontophoresis was performed using commercially available patches (10 cm2 contact area). Following i.v.-bolus, C(max) in skin occurred with a delay of 38 +/- 4 min compared with plasma. No quantifiable concentration of ACV was detected in the skin on passive drug delivery. Following iontophoresis, skin exposure to ACV was 40, 22, and 11% of that following i.v.-bolus. Conversely, systemic exposure to ACV was negligible and plasma concentrations were below the limit of quantification at any time-point. In skin dialysate, C(max), AUC, and half-life increased with current density. During ointment application, ACV in dermis was detectable only for the first 30 min thereafter ACV skin concentrations were below the LOQ (30 ng/ml).     

Elevated maternal mid-trimester chorionic gonadotropin > or =4 MoM is associated with fetal cerebral blood flow redistribution

BACKGROUND: Elevated mid-trimester human chorionic gonadotropin (hCG) is associated with adverse maternal and perinatal outcome. The aims of the study were to evaluate the association between elevated hCG, fetal pathological arterial waveforms and maternal and perinatal complications. METHODS: Pulsatility indices (PI) of middle cerebral artery (MCA) and umbilical artery (UA) were determined prospectively in 121 consecutive patients with abnormal maternal serum hCG (> 2.5 MoM). Each patient had four US scans during pregnancy. Patients with known structural or chromosomal anomalies were excluded. RESULTS: Of 121 women with hCG > 2.5 MoM, 36/121(29.6%) had hCG between 2.5 and 3.0 MoM, 35/121(28.9%) had hCG between 3.0 and 3.5 MoM, 21/121(17.3%) had hCG of 3.5-4.0 MoM, 17/121(14.1%) had hCG levels between 4.0 and 4.5 MoM, and 12/121(9.9%) had hCG > 4.5 MoM. Middle cerebral artery PI was significantly lower in women with hCG > 4.0 MoM between 28 and 36 weeks' gestation, but not between 18 and 27 weeks' gestation. No differences of MCA PI were found when the cut-off point of hCG was 3.5. Women with hCG levels > 4.0 MoM had a significantly higher rate of preterm deliveries, cesarean sections, higher rate of Apgar scores < 7 and a significantly lower mean birth weight in comparison with women with hCG < 4.0 MoM. The prevalence of PIH and preeclampsia and perinatal death were found to be higher among patients with hCG levels > 4.0 MoM, although not significantly. No differences were found at hCG levels less than 4.0 MoM. CONCLUSIONS: In pregnancies with mid-trimester hCG > 4.0 MoM, redistribution of cerebral blood flow is expressed after 28 weeks' gestation. These pregnancies have higher rates of maternal and neonatal complications as compared to pregnancies with lower hCG levels.     

Presenilin-dependent gamma-secretase activity mediates the intramembranous cleavage of CD44

CD44 is the major adhesion molecule for the extracellular matrix components and is implicated in a wide variety of physiological and pathological processes including the regulation of tumor cell growth and metastasis. Our previous studies have shown that CD44 undergoes sequential proteolytic cleavages in the extracellular and transmembrane domains and the cleavage product derived from CD44 intramembranous cleavage acts as a signal transduction molecule. However, the underlying mechanism of the intramembranous cleavage of CD44 remains to be elucidated. In the present study, we report for the first time that CD44 is a substrate of the presenilin (PS)-dependent gamma-secretase. We demonstrate that the intramembranous cleavage of CD44 induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) treatment or mechanical scraping is blocked by gamma-secretase inhibitors in U251MG cells and that this cleavage is also inhibited in PS-deficient mouse embryonic fibroblasts. Furthermore, we showed that PS1 is redistributed to ruffling areas of the plasma membrane similarly to CD44 after TPA treatment, supporting our biochemical observation that PS1 is involved in the intramembranous cleavage of CD44. Our present findings suggest important implications for understanding CD44-dependent signal transduction and a potential role of PS/gamma-secretase activity in the functional regulation of adhesion molecules.     

Combination drug regimens for metastatic clear cell renal cell carcinoma

Renal cell carcinomas (RCC) make up about 90% of kidney cancers, of which 80% are of the clear cell subtype. About 20% of patients are already metastatic at the time of diagnosis. Initial treatment is often cytoreductive nephrectomy, but systemic therapy is required for advanced RCC. Single agent targeted therapies are moderately toxic and only somewhat effective, leading to development of immunotherapies and combination therapies. This review identifies limitations of monotherapies for metastatic renal cell carcinoma, discusses recent advances in combination therapies, and highlights therapeutic options under development. The goal behind combining various modalities of systemic therapy is to potentiate a synergistic antitumor effect. However, combining targeted therapies may cause increased toxicity. The initial attempts to create therapeutic combinations based on inhibition of the vascular endothelial growth factor or mammalian target of rapamycin pathways were largely unsuccessful in achieving a profile of increased synergy without increased toxicity. To date, five combination therapies have been approved by the U.S. Food and Drug Administration, with the most recently approved therapies being a combination of checkpoint inhibition plus targeted therapy. Several other combination therapies are under development, including some in the phase 3 stage. The new wave of combination therapies for metastatic RCC has the potential to increase response rates and improve survival outcomes while maintaining tolerable side effect profiles.     

Prevalence and Associated Factors of Sarcopenia in Singaporean Adults—The Yishun Study

ObjectivesTo describe the normative values of sarcopenia among community-dwelling adults (≥21 years of age); compare the prevalence of sarcopenia using Asian Working Group for Sarcopenia criteria, 2014 (AWGS2014), Asian Working Group for Sarcopenia criteria, 2019 (AWGS2019), and European Working Group on Sarcopenia in Older People criteria, 2018 (EWGSOP2) guidelines; and identify factors associated with sarcopenia.DesignParticipants were recruited through random sampling. Sarcopenia assessments were performed using a dual-energy x-ray absorptiometry scan (muscle mass), handgrip test (muscle strength), and usual walking test (physical performance). Questionnaires were administered to evaluate lifestyle and cognition.Setting and ParticipantsIn total, 542 community-dwelling Singaporeans were recruited (21?90 years old, 57.9% women).MethodsWe assessed anthropometry, body composition, and questionnaire-based physical and cognitive factors, and estimated sarcopenia prevalence according to the AWGS2014, AWGS2019, and EWGSOP2 recommendations, and examined associations using logistic regression.ResultsAccording to AWGS2019, the Singapore population-adjusted sarcopenia prevalence was 13.6% (men 13.0%; women 14.2%) overall, and 32.2% (men 33.7%, women 30.9%) in those aged 60 years and above. The cut-offs derived from young adult reference group for low appendicular lean mass index were 5.28 kg/m2 for men and 3.69 kg/m2 for women (lower than AWGS recommended cut-off); for gait speed it was 0.82 m/s, (AWGS2019 recommended cut-off 1.0 m/s, AWGS2014 cut-off was 0.8 m/s); and for handgrip strength it was 27.9 kg/m2 for men and 16.7 kg/m2 for women (close to AWGS2019 recommendation). Age, sex, marital status, alcoholism, physical activity, body mass index, waist circumference, and global cognition were associated with sarcopenia (P < .05).Conclusions and ImplicationsThis is the first study to provide reference values of muscle mass, strength, and gait speed across the adult lifespan of Singaporeans. Using AWGS2019 criteria, sarcopenia is prominent in older age (32.2% in ≥60 years old), but it is already nontrivial (6.9%) among young and middle-age persons. Multidomain lifestyle modifications addressing muscle strength, cognition, and nutrition over the adult lifespan are important to delay the development of sarcopenia.     

Understanding Utilization of Outpatient Clinics for Children with Special Health Care Needs in Southern Israel

To understand the pattern of utilization of ambulatory care by parents of children with special health care needs (CSHCN) and to explore parental challenges in coping with health maintenance of their infants after discharge from a neonatal intensive care unit (NICU). CSHCN require frequent utilization of outpatient ambulatory clinics especially in their first years of life. Multiple barriers are faced by families in disadvantaged populations which might affect adherence to medical referrals. Our study attempts to go beyond quantitative assessment of adherence rates, and capture the influence of parental agency as a critical factor ensuring optimal utilization of healthcare for CSHCN. A prospective, mixed-methods, cohort study followed 158 Jewish and Bedouin-Arab infants in the first year post discharge from NICU in southern Israel. Rates of utilization of ambulatory clinics were obtained from medical records, and quantitative assessment of factors affecting it was based on structured interviews with parents at baseline. Qualitative analysis was based on home visits or telephone in-depth interviews conducted about 1 year post-discharge, to obtain a rich, multilayered, experiential perspectives and explained perceptions by parents. Adherence to post-discharge referrals was generally good, but environmental, cultural, and financial obstacles to healthcare, magnified by communication barriers, forced parents with limited resources to make difficult choices affecting utilization of healthcare services. Improving concordance between primary caregivers and health care providers is crucial, and further development of supportive healthcare for CSHCN in concordance with parental limitations and preferences is needed.