Modulation of intestinal TLR4 expression in infants with neonatal opioid withdrawal syndrome.

OBJECTIVE: Neonatal Opioid Withdrawal Syndrome (NOWS) has been associated with the development of necrotizing enterocolitis (NEC) in term and late-preterm neonates. In this study, we used stool gene expression to determine if an increase in baseline inflammation in the intestine of infants with NOWS is associated with these findings. STUDY DESIGN: Stool samples were prospectively collected between days 1-3 and days 4-9 after delivery for opioid-exposed ( n = 9) or non-exposed neonates (n = 8). Stool gene expression for TLR4 and HMGB1 was determined via real-time PCR. RESULTS: TLR4 expression was higher in the stool of the non-exposed group in both time periods, between days 1-3 (P < 0.0001) and days 4-9 (P < 0.05) after delivery. No significant difference in HMGB1 expression was found at either time point (P > 0.05). CONCLUSION: These findings point to an important interplay between opioid exposure and/or NOWS and the inflammatory milieu of the neonatal intestine.

Adverse Perinatal Outcomes Associated with Increasing Maternal Obesity.

OBJECTIVE: The aim of this study was to determine adverse perinatal outcomes related to maternal preconception body mass index (BMI). STUDY DESIGN: This is a retrospective observational cohort study at a single institution of 500 consecutive mothers of normal weight with a preconception BMI of 18.5 to less than 25 and 500 additional obese mothers with a preconception BMI more than or equal to 30. Maternal/newborn metrics were stratified by maternal preconception BMI and trend analysis was performed both by simple univariable and multivariable logistic regression analysis. RESULTS: The study included 858 mother/baby dyads after 142 were excluded. Trend analysis demonstrated higher preconception BMI was significantly associated with progressively higher rates of cesarean section (p < 0.001), preeclampsia p < 0.001), gestational diabetes (p < 0.001), preterm birth (p = 0.001), lower 1- and 5 minutes Apgar scores (p < 0.001), and neonatal intensive care unit admission (p = 0.002). These associations remained significant in both simple univariable and multivariable logistic regression models. CONCLUSION: We demonstrated obese women are more likely to have maternal complications and neonatal morbidity when compared with normal weight mothers. Maternal and fetal complications increase with increasing obesity with superobese mothers (BMI ≥ 50) having more perinatal adverse outcomes when compared with other classes of obesity. It is reasonable to counsel weight loss prior to conception of women with BMI more than or equal to 30 in an effort to reduce maternal complications and neonatal morbidity related to pregnancy. KEY POINTS: · Maternal obesity is associated with adverse outcomes.. · Complications increase with increasing obesity.. · Superobese mothers have the most adverse outcomes..

Examination of cord blood at birth in women with SARS-CoV-2 exposure and/or vaccination during pregnancy and relationship to fetal complete blood count, cortisol, ferritin, vitamin D, and CRP.

Background: SARS-CoV-2 is known to manifest a robust innate immune response. However, little is known about inflammatory influences from maternal SARS-CoV-2 infection or maternal mRNA vaccination upon the fetus. In addition, it is unknown if Vitamin D deficiency influences fetal homeostasis or if an anti-inflammatory mechanism to the development of possible innate cytokines or acute phase reactants by the maternal/fetal dyad, in the form of cortisol elevations, occur. In addition, effects on Complete Blood Count (CBC) are not known. Objective: To evaluate the neonatal acute phase reactants and anti-inflammatory responses after maternal SARS-CoV-2 disease or mRNA vaccination. Methods: Samples and medical records reviews from mother/baby dyads (n = 97) were collected consecutively, and were categorized into 4 groups; no SARS-CoV-2 or vaccination exposure (Control), Vaccinated mothers, maternal SARS-CoV-2 disease positive/IgG titer positive fetal blood, and maternal SARS-CoV-2 positive/IgG titer negative fetal blood. SARS-CoV-2 IgG/IgM/IgA titers, CBC, CRP, ferritin, cortisol, and Vitamin D were obtained to examine the possible development of an innate immune response and possible anti-inflammatory response. Student's t-test, Wilcoxon rank-sum, and Chi-squared with Bonferroni corrections were used to compare groups. Multiple imputations were performed for missing data. Results: Cortisol was higher in babies of both mothers who were vaccinated (p = 0.001) and SARS-CoV-2 positive/IgG positive (p = 0.009) as compared to the control group suggesting an attempt to maintain homeostasis in these groups. Measurements of ferritin, CRP, and vitamin D did not reach statistical significance. CBC showed no variation, except for the mean platelet volume (MPV), which was elevated in babies whose mothers were vaccinated (p = 0.003) and SARS-CoV-2 positive/IgG positive (p = 0.007) as compared to the control group. Conclusion: Acute phase reactant elevations were not noted in our neonates. Vitamin D levels were unchanged from homeostatic levels. Cord blood at birth, showed Cortisol and MPV higher in vaccinated and SARS-CoV-2 IgG positive mother/baby dyads as compared to the Control group, indicating that possible anti-inflammatory response was generated. The implication of possible inflammatory events and subsequent cortisol and/or MPV elevation effects upon the fetus after SARS-CoV-2 disease or vaccination is unknown and merits further investigation.

Carbohydrate-Restricted Diet: A Successful Strategy for Short-Term Management in Youth with Severe Obesity-An Observational Study.

Background: Obesity affects ∼20% of children in the United States and reports of successful dietary treatment are lacking. This study aimed to determine the change in body weight in severely obese youth after carbohydrate-restricted dietary intervention. Methods: This single-center study of a carbohydrate-restricted diet (≤30 grams per day), with unlimited calories, fat, and protein for 3-4 months, examined two groups of severely obese youth of ages 5-18 years: Group A, retrospectively reviewed charts of severely obese youth referred to the Pediatric Obesity Clinic at Hoops Family Children's Hospital and the Ambulatory Division of Marshall Pediatrics, Marshall University School of Medicine, in Huntington, WV, between July 1, 2014 and June 30, 2017 (n = 130), and Group B, prospective participants, referred between July 1, 2018 and December 31, 2018, followed with laboratory studies pre- and postdietary intervention (n = 8). Results: In Group A, 310 participants began the diet, 130 (42%) returned after 3-4 months. Group B had 14 enrollees who began the diet, and 8 followed up at 3-4 months (57%). Girls compared with boys were more likely to complete the diet (P = 0.02). Participants <12 years age were almost twice as likely to complete the diet compared with those 12-18 years (64% vs. 36%, P < 0.01); however, the older group subjects who completed the diet had the same percentage of weight loss compared with those <12 years (6.9% vs. 6.9%). Group A had reductions in weight of 5.1 kg (P < 0.001), body mass index (BMI) 2.5 kg/m2 (P < 0.001), and percentage weight loss 6.9% (P < 0.001). Group B had reductions in weight 9.6 kg (P < 0.01), BMI 4 kg/m2 (P < 0.01), and percentage weight loss 9% (P < 0.01). In addition, participants had significant reductions of fasting serum insulin (P < 0.01), triglycerides (P < 0.01), and 20-hydroxyeicosatetraenoic acid (P < 0.01). Conclusions: This study demonstrated a carbohydrate-restricted diet, utilized short term, effectively reduced weight in a large percentage of severely obese youth, and can be replicated in a busy primary care office.

Better Outcomes for Hospitalized Children through Safe Transitions: A Quality Improvement Project.

This project's goal was to implement an already validated pediatric discharge toolkit to enhance the effectiveness of transition from hospital to home, thus reducing 30-day readmission rates. METHODS: This quality improvement study involved implementing a pediatric discharge planning toolkit to improve upon predetermined outcome measures. Critical elements in the toolkit included: (1) comprehensive patient risk assessment on admission; (2) teach-back curriculum; (3) fax or phone call to the primary care physician; (4) 72-hour follow-up calls; and (5) follow-up appointments, scheduled before discharge, within 2 weeks from discharge from hospital. We used the toolkit to gather data on pediatric patients as they were admitted and then prepare them for discharge from December 2016 until March 2017. The primary outcome measure was the 30-day readmissions to the hospital, and the secondary outcome measure was patient satisfaction scores. Our balancing metrics included follow-up appointments made and length of stay. These measures were compared with preintervention hospital pediatric administrative data collected from December 2015 through March 2016. RESULTS: Data collected during the study period (n = 91) compared to preintervention hospital administrative data collected the year prior (n = 132) showed a 31% reduction in readmissions, 4.8% and 7%, respectively (95% confidence interval 0.68-3.8), P = 0.004. Patient satisfaction scores showed no statistical significance. All patients (100%) in both groups had follow-up appointments made before discharge, and the length of stay showed no statistical difference. CONCLUSIONS: This pediatric discharge toolkit improved the efficacy of transition from hospital to home by reducing 30-day readmissions. Patient satisfaction scores were not reduced by utilizing the toolkit.

Persistent Hypoglycemia with Polycystic Kidneys: A Rare Combination - A Case Report.

We present the case of an infant referred to our NICU born at 39 weeks' gestation with persistent hypoglycemia with elevated insulin levels (HI) requiring diazoxide to maintain normoglycemia. Additionally, polycystic kidney disease (PKD) was detected by ultrasound. Molecular genetic testing revealed pathogenic variants in the PMM2gene, i.e., a variant in the promoter region and a missense variant in the coding region. The precoding variant was recently described in 11 European families with similar phenotypes, either in a homozygous state or as compound heterozygous with a pathogenic coding variant. In neonates with HI associated with PKD, this rare recessive disorder should be considered.

Colon cleansing protocol in children: research conditions vs. clinical practice.

BACKGROUND AND STUDY AIMS: Colon preparation rates are the limiting factor for a successful diagnostic colonoscopy in children. Different colon cleansing protocols have been published for use in children. Unfortunately, the applicability of those published research protocols has not been formally evaluated in routine clinical practice. We investigated the success rate of our previously published colon cleansing protocol as utilized in our clinical practice. PATIENTS AND METHODS: This was a retrospective study. In the clinical practice, the colon cleansing protocol included PEG-3350 at a dose of 2 g/kg/day plus Dulcolax (Bisacodyl, Boehringer Ingelheim, TX USA) 5 mg/day for 2 days. Adequate colon preparation was graded between 1 - 5, as previously described, and grade ≥ 4.0 was considered an adequate preparation. Patients were instructed to complete a questionnaire that included PEG-3350 dose, number of stools per day, consistency of each stool, and side effects (vomiting, abdominal pain). Clinical and endoscopic results were compared between the protocol under research conditions and routine practice. RESULTS: The success rate of the colon preparation in our clinical practice was similar to the results observed under our research protocol (75 % vs. 73.6 %). Moreover, the total number of stools, stool consistency, and the intubation rate of the terminal ileum were also similar. We concluded, that in our experience, the colon cleansing protocol used under research conditions was effective and appropriate for use in routine clinical practice. CONCLUSION: We recommend testing each new protocol under the routine conditions of clinical practice to confirm its applicability for general practitioners.

Tissue Transglutaminase Levels Are Not Sufficient to Diagnose Celiac Disease in North American Practices Without Intestinal Biopsies.

BACKGROUND: Celiac serology is crucial for the diagnosis of celiac disease in children. The American guideline for celiac disease in children suggested that positive serology should be followed by confirmatory intestinal histology. The relationship between high tissue transglutaminase titers and celiac disease in children has not been well investigated in children from North America. AIMS: In the present study, we investigated whether different tissue transglutaminase titers in symptomatic children could predict celiac disease without the confirmation of intestinal histology. METHODS: Data from biopsy confirmed celiac children were collected from four different clinics in North America. Clinical, serological, histological, and follow-up data were collected. The accuracy rates of various tissue transglutaminase titers to predict celiac disease in children were calculated. RESULTS: The data from 240 children were calculated. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy rate of tissue transglutaminase titers at ≥10× upper limit of normal were 75.4, 48.8, 87.7, 29.0, and 70.8 %, respectively. Similar data were noted in the other tissue transglutaminase titers (≥3× upper limit of normal, >100 U/ml, or >100 U/ml and >10× upper limit of normal). CONCLUSIONS: The positive predictive value of tissue transglutaminase titers at ≥3× upper limit of normal or higher was too low to predict celiac disease in children. Our data suggested that in routine clinical practice, high titers of tissue transglutaminase are not sufficient to diagnose celiac disease in North American children without intestinal biopsies.

Role of Serum Biomarkers in Early Detection of Non-Alcoholic Steatohepatitis and Fibrosis in West Virginian Children.

BACKGROUND: Obesity, an epidemic among West Virginia children, as well as insulin resistance (IR), is well-established contributors to nonalcoholic steatohepatitis (NASH). Progression of NASH can lead to hepatic fibrosis and cirrhosis, making early detection imperative. The standard for diagnosing NASH is histologically via liver biopsy, which is highly invasive and generally contraindicated in children. By studying serum biomarkers associated with NASH, we aim to identify high risk children who can benefit from a less invasive, alternative approach to the early detection of NASH. METHODS: Seventy one children were prospectively recruited and divided into 3 groups: normal weight without IR (control), obese without IR, and obese with IR. Serum samples were drawn for each patient and biomarker levels were assessed via ELISA kits. RESULTS: Obese without IR and obese with IR patients had significantly elevated levels of lipid metabolism and accumulation markers (FGF-21, NEFA, FATP5, ApoB), oxidative stress markers (dysfunctional HDL, 8-Isoprostane), inflammatory markers(dysfunctional HDL, CK-18) and apoptosis markers (CK-18) compared to control patients (p<0.02). Bilirubin (an antioxidant) was significantly decreased in the obese without IR and obese with IR patients compared to control (p<0.02). CONCLUSION: This study showed a correlation between obesity, IR, and biomarkers associated with NASH in pediatrics patients from West Virginia, with obese with IR patients showing the strongest correlation. These findings support the clinical application of these serum biomarkers as a less invasive method for early detection of NASH and hepatic fibrosis.

Rate of celiac disease in children: view from the endoscopy suite.

OBJECTIVES: The low rate of celiac disease diagnosed in children from the United States may be limited by the practice of "serology-led" diagnosis. The frequency of seronegative celiac disease is unknown, but is underestimated in children and may result in misdiagnosis of celiac disease. The aim of the present study was to investigate the rate of celiac disease after upper endoscopy (esophagogastroduodenescopy [EGD]) with no prior positive celiac serology compared with the rate of celiac disease followed by positive serology. METHODS: Charts of all of the first diagnostic EGDs in children (2009-2013) were retrospectively reviewed. Patients with confirmed celiac disease were divided into 4 groups: group A, positive EGD/positive serology (histology-led diagnosis); group B, positive serology/positive histology (serology-led diagnosis); group C, positive histology followed by negative serology (control 1); and group D, positive serology followed by negative histology (control 2). RESULTS: A total of 761 upper endoscopic charts were reviewed. Of these, 15 children were confirmed with celiac disease (1.97%). There was no significant difference in the demographic data or clinical symptoms between group A and group B. No significant difference was observed in the rate of celiac disease between histology-led celiac diagnosis (group A) and serology-led celiac diagnosis (group B) (1.18% vs 0.79%, P = 0.273). CONCLUSIONS: The rate of celiac disease in endoscopy-led diagnosis was comparable to that in the serology-led diagnosis, suggesting that to increase the detection of celiac disease in children, an adequate number of intestinal biopsies should be performed in every diagnostic upper endoscopic procedure.

4-Amino-2-chlorophenol: Comparative in vitro nephrotoxicity and mechanisms of bioactivation.

Chlorinated anilines are nephrotoxicants both in vivo and in vitro. The mechanism of chloroaniline nephrotoxicity may occur via more than one mechanism, but aminochlorophenol metabolites appear to contribute to the adverse in vivo effects. The purpose of this study was to compare the nephrotoxic potential of 4-aminophenol (4-AP), 4-amino-2-chlorophenol (4-A2CP), 4-amino-3-chlorophenol (4-A3CP) and 4-amino-2,6-dichlorophenol (4-A2,6DCP) using isolated renal cortical cells (IRCC) from male Fischer 344 rats as the model and to explore renal bioactivation mechanisms for 4-A2CP. For these studies, IRCC (∼4×10(6)cells/ml) were incubated with an aminophenol (0.5 or 1.0mM) or vehicle for 60min at 37°C with shaking. In some experiments, cells were pretreated with an antioxidant or cytochrome P450 (CYP), flavin-containing monooxygenase (FMO), peroxidase or cyclooxygenase inhibitor prior to 4-A2CP (1.0mM). Lactate dehydrogenase (LDH) release served as a measure of cytotoxicity. The order of decreasing nephrotoxic potential in IRCC was 4-A2,6-DCP>4-A2CP>4-AP>4-A3CP. The cytotoxicity induced by 4-A2CP was reduced by pretreatment with the peroxidase inhibitor mercaptosuccinic acid, and some antioxidants (ascorbate, glutathione, N-acetyl-l-cysteine) but not by others (α-tocopherol, DPPD). In addition, pretreatment with the iron chelator deferoxamine, several CYP inhibitors (except for the general CYP inhibitor piperonyl butoxide), FMO inhibitors or indomethacin (a cyclooxygenase inhibitor) failed to attenuate 4-A2CP cytotoxicity. These results demonstrate that the number and ring position of chloro groups can influence the nephrotoxic potential of 4-aminochlorophenols. In addition, 4-A2CP may be bioactivated by cyclooxygenase and peroxidases, and free radicals appear to play a role in 4-A2CP cytotoxicity.

3,4,5-Trichloroaniline nephrotoxicity in vitro: potential role of free radicals and renal biotransformation.

Chloroanilines are widely used in the manufacture of drugs, pesticides and industrial intermediates. Among the trichloroanilines, 3,4,5-trichloroaniline (TCA) is the most potent nephrotoxicant in vivo. The purpose of this study was to examine the nephrotoxic potential of TCA in vitro and to determine if renal biotransformation and/or free radicals contributed to TCA cytotoxicity using isolated renal cortical cells (IRCC) from male Fischer 344 rats as the animal model. IRCC (~4 million cells/mL; 3 mL) were incubated with TCA (0, 0.1, 0.25, 0.5 or 1.0 mM) for 60-120 min. In some experiments, IRCC were pretreated with an antioxidant or a cytochrome P450 (CYP), flavin monooxygenase (FMO), cyclooxygenase or peroxidase inhibitor prior to incubation with dimethyl sulfoxide (control) or TCA (0.5 mM) for 120 min. At 60 min, TCA did not induce cytotoxicity, but induced cytotoxicity as early as 90 min with 0.5 mM or higher TCA and at 120 min with 0.1 mM or higher TCA, as evidenced by increased lactate dehydrogenase (LDH) release. Pretreatment with the CYP inhibitor piperonyl butoxide, the cyclooxygenase inhibitor indomethacin or the peroxidase inhibitor mercaptosuccinate attenuated TCA cytotoxicity, while pretreatment with FMO inhibitors or the CYP inhibitor metyrapone had no effect on TCA nephrotoxicity. Pretreatment with an antioxidant (α-tocopherol, glutathione, ascorbate or N-acetyl-L-cysteine) also reduced or completely blocked TCA cytotoxicity. These results indicate that TCA is directly nephrotoxic to IRCC in a time and concentration dependent manner. Bioactivation of TCA to toxic metabolites by CYP, cyclooxygenase and/or peroxidase contributes to the mechanism of TCA nephrotoxicity. Lastly, free radicals play a role in TCA cytotoxicity, although the exact nature of the origin of these radicals remains to be determined.

Does Helicobacter pylori protect against eosinophilic esophagitis in children?

BACKGROUND: Helicobacter pylori infection and eosinophilic esophagitis (EoE) in children seem to have a reversed association with socioeconomic status (hygienic condition) and allergy conditions. While Hp infection (Hp) is highly associated with poor hygiene and/or poor socioeconomic status, but not with allergic conditions (asthma, rhinitis, etc.), EoE has the opposite epidemiological relationship (high association with allergy but low with low hygienic conditions). AIM: To investigate the association between Hp infection and EoE in children. METHODS: A retrospective chart review of all children who undergo the first upper endoscopy procedure in the gastroenterology clinic, between 2007 and 2012, was performed. Demographic, endoscopic and histological data were collected. The data was divided into 4 diagnostic groups: Hp infection, EoE, reflux esophagitis, and children who had normal histology. The relationship between Hp positive children and the other groups was performed. RESULTS: A total of 966 charts were available for review. Esophagitis, idiopathic gastritis, EoE, and Hp infection were detected in 268 (28%), 480 (49%), 62 (6%), and 31 (3%) children, respectively. The mean age of the EoE group was significantly lower compared to all reference groups (p < .002), but no significant different was detected among the reference groups (gastritis, GERD, and Hp infection; p = 1.00). Simple logistic regression analysis using Hp infection as a predictor for EoE did not find a significant relationship between these two variables (p-value = .471, OR = 0.478, 95% CI 0.06-3.56). However, multivariable logistic regression analysis between EoE and the reference groups indicated a significant negative relationship between Hp infection and EoE (p-value = .023, adjusted OR = 0.096, 95%CI 0.013-0.72). Neither gastritis nor GER showed significant relationship with EoE (p-values are 1.000 and .992, respectively). CONCLUSION: A reversed association between Hp and EoE was found in a cohort of West Virginia children. The possible explanations for these findings are discussed.

Upper endoscopy in children: do symptoms predict positive findings?

Upper endoscopy (esophagogastroduodenoscopy or EGD) is an important diagnostic tool for many gastrointestinal symptoms. In recent years, the number of EGDs has increased dramatically. Unfortunately, the rate of negative (normal) EGD in children is high, approximating 50% of all procedures. To decrease the cost of EGD procedures, it is important to assess which clinical symptom would detect positive findings. This information may also be valuable in improving the referral practices of the primary care physicians for EGD. In a retrospective study, we investigated the pathological yield of the first EGD in children referred for various symptoms. Abdominal pain was the most common referral symptom and the best predictor of positive EGD, reaching an accuracy level of 79.9%. All other investigated symptoms had less than 50% accuracy. We concluded that most gastrointestinal symptoms in children have a poor predictive value for positive EGD. A cost-benefit analysis of EGD in children is needed.

A review of the efficacy of the picture exchange communication system intervention.

The Picture Exchange Communication System (PECS) is a communication program that has become widely used, especially with children with autism. This paper reports the results of a review of the empirical literature on PECS. A descriptive review is provided of the 27 studies identified, which included randomized controlled trials (RCTs), other group designs and single subject studies. For 10 appropriate single subject designs the percentage of nonoverlapping data (PND) and percentage exceeding median (PEM) metrics were examined. While there are few RCTs, on balance, available research provides preliminary evidence that PECS is readily learned by most participants and provides a means of communication for individuals with little or no functional speech. Very limited data suggest some positive effect on both social-communicative and challenging behaviors, while effects on speech development remain unclear. Directions for future research are discussed including the priority need for further well-conducted RCTs.

The relationship of stigma to the sexual risk behavior of rural men who have sex with men.

Most studies of risky sexual behaviors of men who have sex with men (MSM) have been conducted in cities. Few have documented risky sexual behavior of rural men despite increases in rural HIV. Fewer have addressed stigma and risk. This study explored the effects of stigma on sexual risk behavior among rural MSM. We hypothesized that stigma emanating from families, health care providers, and the communities of rural MSM would indirectly affect their sexual risk behavior through their mental health status, specifically self-esteem and internalized homophobia. A convenience sample of 414 rural MSM obtained through political, health service, and social organizations completed an anonymous self-administered questionnaire. Over half of the men reported high-risk sexual behavior. Sensation seeking directly affected levels of sexual risk while the effects of stigma on sexual risk behavior were mediated by mental health variables. Stigma related to respondents' low self-esteem, and low internalized homophobia increased risk behavior.

Uniaxial stretch-induced regulation of mitogen-activated protein kinase, Akt and p70 S6 kinase in the ageing Fischer 344 x Brown Norway rat aorta.

The effects of ageing on the cardiovascular system contribute to substantial alterations in cellular morphology and function. The variables regulating these changes are unknown; however, one set of signalling molecules that may be of particular importance in mediating numerous cellular responses, including control of cell growth, differentiation and adaptation, are the proteins associated with the mitogen-activated protein kinase (MAPK) signalling systems. The MAPKs, in conjunction with the p70 S6k signalling cascade, have emerged as critical components for regulating numerous mechanotransduction-related cellular responses. Here we investigate the ability of uniaxial stretch to activate the MAPK and p70 S6k pathways in adult (6-month-old), aged (30-month-old) and very aged (36-month-old) Fischer 344/NNiaHSd x Brown Norway/BiNia (FBN) rats. Western blotting of the MAPK family proteins extracellular signal-regulated kinase (Erk) 1/2, p38- and c-Jun NH(2)-terminal kinase (Jnk)-MAPKs showed differential expression and activation between these proteins with age. An acute 15 min interval of 20% uniaxial stretch using an ex vivo aortic preparation demonstrated similar regulation of Erk1/2, p38- and Jnk-MAPK. However, ageing altered uniaxial induced p70 S6k pathway signalling. These observations confirm previous data demonstrating that MAPK proteins are mechanically regulated and also suggest that p70 S6k signalling expression and activation are controlled differently with ageing. Taken together, these data may help to explain, in part, the age-related changes in vascular morphology, function and response to injury.

Age-related dystrophin-glycoprotein complex structure and function in the rat extensor digitorum longus and soleus muscle.

This study tested the hypothesis that age-related changes in the dystrophin-glycoprotein complex (DGC) may precede age-associated alterations in muscle morphology and function. Compared to those in adult (6 month) rats, extensor digitorum longus (EDL) and soleus muscle mass was decreased in old (30 month) and very old (36 month) Fischer 344/NNiaHSD x Brown Norway/BiNia rats. The amount of dystrophin, beta-dystroglycan, and alpha-sarcoglycan increased with aging in the EDL and decreased with aging in the soleus. alpha-Dystroglycan levels were increased with aging in both muscles and displayed evidence of altered glycosylation. Immunostaining for the presence of antibody infiltration and dystrophin following increased muscle stretch suggested that the aging in the soleus was characterized by diminished membrane integrity. Together, these data suggest that aging is associated with alterations in EDL and soleus DGC protein content and localization. These results may implicate the DGC as playing a role in age-associated skeletal muscle remodeling.

Diabetes alters vascular mechanotransduction: pressure-induced regulation of mitogen activated protein kinases in the rat inferior vena cava.

BACKGROUND: Diabetes mellitus is an important risk factor for increased vein graft failure after bypass surgery. However, the cellular and molecular mechanism(s) underlying vessel attrition in this population remain largely unexplored. Recent reports have suggested that the pathological remodeling of vein grafts may be mediated by mechanically-induced activation of the mitogen activated protein kinase (MAPK) signaling pathways and the MAPK-related induction of caspase-3 activity. On the basis of these findings, we hypothesized that diabetes may be associated with alterations in how veins "sense" and "respond" to altered mechanical loading. METHODS: Inferior venae cavae (IVC) from the non-diabetic lean (LNZ) and the diabetic obese (OSXZ) Zucker rats were isolated and incubated ex vivo under basal or pressurized conditions (120 mmHg). Protein expression, basal activation and the ability of increased pressure to activate MAPK pathways and apoptosis-related signaling was evaluated by immunoblot analysis. RESULTS: Immunoblot analyses revealed differential expression and activation of extracellular signal-regulated kinase (ERK1/2), p38 and c-Jun NH2-terminal kinase (JNK) MAPKs in the IVCs of diabetic rats as compared to non-diabetic rats. In particular, the expression and basal phosphorylation of p38beta- (52.3 +/- 11.8%; 45.8 +/- 18.2%), JNK 1- (21.5 +/- 9.3%; 19.4 +/- 11.6%) and JNK3-MAPK (16.8 +/- 3.3%; 29.5 +/- 17.6%) were significantly higher (P < 0.05) in the diabetic vena cava. An acute increase in IVC intraluminal pressure failed to increase the phosphorylation of ERK1-, JNK-2, or any of the p38-MAPKs in the diabetic obese Zucker rats. Also, IVC loading in the LNZ led to a 276.0 +/- 36.0% and 85.8 +/- 25.1% (P < 0.05) increase in the cleavage of caspase-3 and caspase-9, respectively, with no effect on these molecules in the OSXZ. No differences were found in the regulation of Bax and Bcl-2 between groups. However, basal expression levels of Akt, phospho-Akt, PTEN, phospho-PTEN and phospho-Bad were higher in the diabetic venae cavae (P < 0.05). CONCLUSION: These data suggest that diabetes is associated with significant alteration in the ability of the vena cava to activate MAPK- and apoptosis-related signaling. Whether these changes are associated with the increased vein graft attrition seen in the diabetic population will require further investigation.