Expression and synthesis of fibroblast growth factor-9 in human gammadelta T-lymphocytes. Response to isopentenyl pyrophosphate and TGF-beta1/IL-15

Gammadelta T-lymphocytes are believed to play a role in maintaining the normal configuration of epithelial tissue. As little is known about the factors mediating this function, we addressed the question of whether gammadelta T-lymphocytes produce fibroblast growth factor (FGF)-9 as well as two other growth factors associated with epithelial tissue reconstitution. Blood gammadelta T cells isolated from healthy donors were grown in the presence of isopentenyl pyrophosphate (IPP) or transforming growth factor-beta1 (TGF-beta1)/interleukin-15 (IL-15) for 24 h and were assessed for the expression and synthesis of FGF-9, keratinocyte growth factor (KGF), and epidermal growth factor (EGF). Resting human gammadelta T cells constitutively expressed KGF and FGF-9 mRNA but no EGF mRNA. In the presence of IPP, FGF-9 mRNA expression significantly increased in a dose-dependent manner, expression of KGF remained unaltered, and EGF mRNA could not be detected. In contrast to IPP, stimulation of the cells with TGF-beta1/IL-15 did not alter FGF-9 expression. Moreover, stimulation with anti-CD3 does not induce FGF-9 expression but triggers a high signal of interferon-gamma mRNA. Western blot analysis of gammadelta T cell lysates, prepared 4 days following stimulation with IPP, showed an increase of FGF-9 protein as compared with control cells. In conclusion, the results demonstrate for the first time that human blood and bronchoalveolar lavage gammadelta T-lymphocytes are capable of expressing FGF-9. The data also provide novel evidence that immunoregulatory cells can synthesize FGF-9.     

Placental telomere length and risk of placental abruption

Objective: To investigate the associations of placental telomere length with placental abruption (PA) risk and interactions between placental telomere length and placental mitochondrial DNA (mtDNA) copy number on PA risk.

Materials and methods: Relative telomere length and mtDNA copy number in placental samples collected from 105 cases and 73 controls were measured in two batches using qRT-PCR. Mean differences in relative telomere length between PA cases and controls were examined. After creating batch-specific median cutoffs for relative telomere length (84.92 and 102.53) and mtDNA copy number (2.32 and 1.42), interaction between the two variables was examined using stratified logistic regression models.

Results: Adjusted mean difference in relative telomere length between PA cases and controls was ?0.07 (p?>?0.05). Among participants with low mtDNA copy number, participants with short relative telomere length had a 3.07-fold higher odds (95% CI: 1.13–8.38) of PA as compared with participants with long relative telomere length (the reference group). Among participants with high mtDNA copy number, participants with short relative telomere length had a 0.71-fold lower odds (95% CI: 0.28–1.83) of PA as compared with the reference group (interaction p values?=?0.03). Conclusion: Findings suggest complex relationships between placental telomere length, mtDNA copy number and PA risk which warrant further larger studies.     

Salivary and Serum Antibody Response Against Neisseria meningitidis After Vaccination With Conjugate Polysaccharide Vaccines in Ethiopian Volunteers

Meningococcal conjugate vaccines induce serum antibodies crucial for protection against invasive disease. Salivary antibodies are believed to be important for hindering meningococcal acquisition and/or clearance of established carriage. In this study, we measured salivary IgA and IgG antibodies induced by vaccination with a monovalent serogroup A conjugate vaccine or a tetravalent A, C, W and Y conjugate vaccine, in comparison with antibody levels in serum. Saliva and serum samples from Ethiopian volunteers (1–29 years) collected before and eight times on a weekly basis after receiving the serogroup A conjugate vaccine, the tetravalent serogroup A, C, W and Y conjugate vaccine, or no vaccine (control group), were analysed using a multiplex microsphere immunoassay for antibody detection. Serogroup‐specific IgG antibody levels in saliva increased significantly after vaccination with both vaccines. The monovalent serogroup A vaccine also induced an increase in salivary IgA antibodies. A strong correlation between serogroup‐specific IgG antibodies in saliva and serum, and a somewhat lower correlation for IgA, was observed for all serogroups. There was also a strong correlation between specific secretory IgA and IgA antibodies in saliva for all serogroups. Meningococcal conjugate vaccines are able to elicit salivary antibodies against serogroup A, C, W and Y correlating with antibody levels in serum. The strong correlation between saliva and serum antibody levels indicates that saliva may be used as a surrogate of systemic antibody responses.     

Genomics of body fat percentage may contribute to sex bias in anorexia nervosa

Anorexia nervosa (AN) occurs nine times more often in females than in males. Although environmental factors likely play a role, the reasons for this imbalanced sex ratio remain unresolved. AN displays high genetic correlations with anthropometric and metabolic traits. Given sex differences in body composition, we investigated the possible metabolic underpinnings of female propensity for AN. We conducted sex‐specific GWAS in a healthy and medication‐free subsample of the UK Biobank (n = 155,961), identifying 77 genome‐wide significant loci associated with body fat percentage (BF%) and 174 with fat‐free mass (FFM). Partitioned heritability analysis showed an enrichment for central nervous tissue‐associated genes for BF%, which was more prominent in females than males. Genetic correlations of BF% and FFM with the largest GWAS of AN by the Psychiatric Genomics Consortium were estimated to explore shared genomics. The genetic correlations of BF%_male and BF%_female with AN differed significantly from each other (p < .0001, δ = ?0.17), suggesting that the female preponderance in AN may, in part, be explained by sex‐specific anthropometric and metabolic genetic factors increasing liability to AN.     

Both membrane-bound and soluble forms of CD14 bind to Gram-negative bacteria

Tissue macrophages and their precursors – the blood monocytes – respond rapidly to a bacterial infection with the release of inflammatory mediators. These mediators are involved in the recruitment of phagocytic cells, principally neutrophils, from the blood to the site of infection. To initiate this process macrophages and monocytes must be able to detect the presence of bacteria in a reliable, but nevertheless nonspecific, fashion. It is thought that this is achieved by means of receptors on the cell surface which recognize structures common to many different bacteria. One candidate for such a “pattern recognition element” is the cell surface glycoprotein CD 14. CD 14 has been shown to bind components of the Gram-positive cell wall and it also binds soluble lipopolysaccharide released from Gram-negative bacteria. In both cases the interaction with CD 14 leads to an activation of the cell. Here we show that human peripheral blood monocytes can, in addition, bind intact Gram-negative bacteria in the presence of serum and that this process involves CD14. When CD14 expression is induced on the myelomonocytic cell line U937 by treatment with vitamin D3 the cells concomittently acquire the capacity to bind bacteria. Furthermore, a non-monocytic cell line which does not bind bacteria acquires the capacity to do so when transfected with either the human or mouse CD14 gene. This binding can be inhibited by blocking the CD14 receptor with anti-CD14 antibody or by blocking the ligand on the bacteria with soluble CD14. Finally we demonstrate binding of sCD14 to Escherichia coli. We conclude that in the presence of serum both membrane-bound and soluble forms of CD14 can bind to Gram-negative bacteria. This suggests that CD14 may play a role in the detection and elimination of intact bacteria in vivo.     

Determination of lamivudine and stavudine in pharmaceutical preparations using chemometrics-assisted spectrophotometry

A simple chemometrics-assisted spectrophotometric method for the simultaneous determination of lamivudine and stavudine in pharmaceutical tablets is described. The UV absorption spectra of the studied drugs, in the range of 200–310 nm, showed a considerable degree of spectral overlapping ([D_i]^0.5 = 94.9%). Resolution of the mixture has been accomplished by using classical least-squares regression analysis (CLS) and principle components regression analysis methods (PCR). Beer’s law was obeyed for both drugs in the general concentration ranges of 2–12 and 3–15 μg ml⁻¹ for lamivudine and stavudine, respectively. The proposed methods were successfully applied for the determination of the two drugs in laboratory prepared mixtures. The overall recoveries percent were found 98.58 ± 1.53–101.30 ± 1.35 (CLS) and 98.62 ± 1.65–101.13 ± 1.04 (PCR) for lamivudine and 98.43 ± 1.62–99.42 ± 1.55 (CLS) and 98.23 ± 1.97–101.20 ± 1.79 (PCR) for stavudine, respectively. The commercial tablets percentage content was found 98.10 ± 2.5–102.47 ± 2.94 (CLS) and 99.12 ± 1.71–100.92 ± 1.54 (PCR) for lamivudine and 96.00 ± 2.94–98.17 ± 1.72 (CLS) and 97.40 ± 1.55–97.80 ± 1.92 (PCR) for stavudine, respectively. Good percentage recoveries and proper statistical data obtained with both the laboratory prepared mixtures and the commercial tablets proved the suitability and efficiency of the proposed procedures for routine analysis and quality control purposes with quite satisfactory precision. A comparison of the obtained results from CLS and PCR were also performed with those obtained from reported method. The obtained F- and t-values obtained indicating no significant differences between the results of the proposed and reported methods.     

Birth Weight,Genetic Susceptibility,and Adulthood Risk of Type 2 Diabetes

OBJECTIVE

Both stressful intrauterine milieus and genetic susceptibility have been linked to later-life diabetes risk. The current study aims to examine the interaction between low birth weight, a surrogate measure of stressful intrauterine milieus, and genetic susceptibility in relation to risk of type 2 diabetes in adulthood.

RESEARCH DESIGN AND METHODS

The analysis included two independent, nested case-control studies of 2,591 type 2 diabetic case subjects and 3,052 healthy control subjects. We developed two genotype scores: an obesity genotype score based on 32 BMI-predisposing variants and a diabetes genotype score based on 35 diabetes-predisposing variants.

RESULTS

Obesity genotype scores showed a stronger association with type 2 diabetes risk in individuals with low birth weight. In low–birth weight individuals, the multivariable-adjusted odds ratio (OR) was 2.55 (95% CI 1.34–4.84) by comparing extreme quartiles of the obesity genotype score, while the OR was 1.27 (1.04–1.55) among individuals with birth weight >2.5 kg (P for interaction = 0.017). We did not observe significant interaction between diabetes genotype scores and birth weight with regard to risk of type 2 diabetes. In a comparison of extreme quartiles of the diabetes gene score, the multivariable-adjusted OR was 3.80 (1.76–8.24) among individuals with low birth weight and 2.27 (1.82–2.83) among those with high birth weight (P for interaction = 0.16).

CONCLUSIONS

Our data suggest that low birth weight and genetic susceptibility to obesity may synergistically affect adulthood risk of type 2 diabetes.Accumulating evidence has shown that the risk of type 2 diabetes in later life might be programmed by intrauterine exposure to environmental stress such as malnutrition (1,2), which may cause physiological, epigenetic, or structural alterations related to poor development of pancreatic β-cell mass and function (3) or insulin resistance (4) in the offspring and subsequently increase the susceptibility to risk of type 2 diabetes during adulthood (1,2). Low birth weight, as a surrogate for prenatal malnutrition, is common in both developing and developed countries, with prevalence reaching 8% in the U.S. and 15.5% worldwide (5). In epidemiology studies, low birth weight has consistently been related to increased diabetes risk (6–8).Recent large-scale genome-wide association studies confirm that common variants in the human genome also contribute to the development of type 2 diabetes (9,10). Thus far, nearly 40 loci have been related to diabetes risk at the genome-wide significance level. In addition, genetic variants predisposing to obesity (11), the most important risk factor for type 2 diabetes, have also been found to be related to diabetes risk (12). These genetic variants may affect disease risk through influencing either β-cell function or insulin resistance.Interestingly, only a few diabetes (13,14) or obesity (15) loci are directly related to birth weight and with quite complex effects: some type 2 diabetes risk alleles are associated with reduced (13) while some others with increased (14) birth weight. Those results suggest that the genetic variants and birth weight may affect the disease risk through different mechanisms. However, the pathways linking low birth weight or genetic variants to diabetes are interwoven. Therefore, we assume that these two types of risk factors may interact in determining risk of type 2 diabetes.In this study, we assessed the potential interaction between birth weight and genetic susceptibility to type 2 diabetes and obesity on risk of type 2 diabetes in two independent prospective cohorts: the Nurses’ Health Study (NHS) and the Health Professionals Follow-up Study (HPFS). The genetic susceptibility was evaluated by combining all the identified common variants from recent genome-wide association studies.     

Interferon-alphacon-1 treatment of three patients with severe glucocorticoid-dependent asthma. Effect on disease control and systemic glucocorticosteroid dose

We report herein the therapeutic effect of interferon (IFN)-alphacon in three patients with severe persistent asthma and long-term oral glucocorticoid treatment. IFN-alphacon (9 microg) administered subcutaneously thrice a week over a period of more than 24 months led to a substantial clinical improvement with regard to the number of daily asthma attacks, nighttime disturbance, emergency visits and hospitalizations. In addition, lung function and exercise capacity improved. At the same time, treatment with IFN allowed discontinuation of the daily glucocorticoid dose in all patients for the first time in more than 8 years. Our findings suggest that IFN-alphacon leads to a significant clinical improvement while at the same time allowing reduction and discontinuation of the glucocorticoid treatment in severe persistent glucocorticoid-dependent asthma.