Haplotypes and mutations of the PAH locus in Egyptian families with PKU

A high degree of molecular heterogeneneity at the phenylalanine hydroxylase (PAH) locus was established by examining RFLP haplotypes and PAH mutations in the families of 13 Egyptians with phenylketenouria (PKU). Thirteen different haplotypes were unequivocally determined in these kindreds. Haplotypes 1.8, 3.9, 4.3, 7.8, 22.11, 27.6, and 52.8 were found segregating with normal chromosomes, whilst haplotypes 1.8, 5.9, 23.8, 32.8, the newly assigned 73.9, and two as yet incomplete but novel haplotypes were found segregating with the mutant chromosomes. There was no particular preference for a single haplotype among normal or mutant chromosomes. Nine different mutations were also identified among the 26 alleles. IVS 10nt11g (8/26), IVS 2nt5g-c (4/26), R261Q (3/26), R176X (2/26), Y206D (2/26), S231P (2/26), Y198fs [593-614del22bp]; (2/26), G46fs [136/137delG]; (1/26), and E178G (1/26). Six of these mutations (IVS 2nt5g-c, R176X, Y198fs, R261Q, S231P, and IVS 10nt11g) are common to other Mediterranean populations. Two mutations not previously reported in the Mediterranean basin were also observed (Y206D and G46fs). These intriguing preliminary findings confirm IVS 10nt11g as a major mutation among Mediterranean mutations and demonstrate the need for a more comprehensive study of Arab populations to confirm the uniqueness of the two novel mutations to the Egyptian population.     

Tc-99m diethylenetriamine pentaacetic acid (DTPA): Is it reliable for assessment of methotrexate-induced cumulative effect on renal filtration in rheumatoid arthritis patients?

Methotrexate[MTX] is commonly employed as the initial DMARD used for treatment of Rheumatoid arthritis[RA]. We aimed to contribute to the safety profile of MTX by assessing its cumulative effect on renal filtration. Fifty two RA adult females with normal base-line serum creatinine and GFR at the initial diagnosis of the disease were included. Group-1[G1] included 30 patients[mean age 40.4 ± 4.4 years] on MTX and NSAIDS, while 22 RA patients[mean age 38.5 ± 8.2 years] who received NSAIDs only served as the control group[G2]. Renal function was assessed by GFR-measurement using Technetium diethylenetriamine-pentaacetic acid[Tc-99 m-DTPA] at the point of the study time corresponding to disease duration. 21/30[70%] in G1 showed reduced GFR compared to 6/22[27.3%] in G2[P0.007] with 3.3 ± 0.5% annual reduction of GFR. Reduced GFR in G1 showed significant negative correlation with age[r = ?0.396, P = 0.005], MTX-cumulative dose[r = ?0.263, P = 0.049], MTX-intake duration[r = ?0.293, P = 0.031] and NSAID-intake duration[r = ?0.344, P = 0.014]. Low dose MTX has a slow cumulative effect on renal filtration manifested by GFR reduction over time that could be monitored by Tc-99 m DTPA.     

“Children are Sexually Innocent”: Iranian Parents’ Understanding of Children’s Sexuality

Sexuality education (SE) is hotly contested in the West and there is much abstinence-only education; however, it remains controversial in a variety of contexts, including in Iran. The lack of consensus about children’s SE in Muslim societies obliges us to explore different aspects of this topic systematically. The qualitative research presented here was about how Iranian parents perceived children’s sexuality. Data from parents of 26 children were collected during four focus group sessions. Informants were selected from Area 5 in West Tehran. This area included 72 primary schools for girls and 82 for boys. The sessions were facilitated by using a semi-structured focus group guide. Content analysis was adopted using combined free and analytical coding to reduce data, to extract meanings, and to categorize domains. One of the three main domains identified, family management of child sexuality, is comprised of the following: (1) understanding of child sexuality, (2) family rules, (3) parent–child interactions, and (4) opposite sex interactions. Parental misinformation, accumulated myths, and ignorance about children’s sexual development were evident. Strict family rules and a lack of consistent policy and instruction for SE were also identified. Parents said they were neither well-prepared nor competent to educate their children about sexuality. In fact, a majority of mothers raised “incompetence” as an important determinant in their own parent–child interactions. Societal changes as well as children’s socialization patterns have forced parents to accept their children’s opposite sex friendships as a reality. Results suggest a community need for developing comprehensive and culturally sensitive SE for schools and parental use.     

Protection by beta-Hydroxybutyric acid against insulin glycation,lipid peroxidation and microglial cell apoptosis

Background

Diabetes mellitus is characterized jointly by hyperglycemia and hyperinsulinemia that make insulin more prone to be glycated and evolve insulin advanced glycation end products (Insulin- AGE). Here, we report the effect of beta-hydroxy butyrate (BHB) (the predominant ketone body) on the formation of insulin-AGE, insulin glycation derived liposomal lipid peroxidation and insulin-AGE toxicity in microglial cells.

Methods

The inhibitory effect of BHB was monitored as a result of insulin incubation in the presence of glucose or fructose using AGE-dependent fluorescence, Tyr fluorescence as well as anilinonaphthalenesulfonate (ANS) andthioflavin T (ThT) binding, and circular dichroism (CD) investigations. To study lipid peroxidation induced by insulin glycation, thiobarbituric acid (TBA) assay and thiobarbituric acid reactive substance (TBARS) monitoring were used. The effect of insulin–AGE on microglial viability was investigated by 3-(4, 5 dimethylthiazol-2-yl)—2, 5-diphenyltetrazoliumbromide (MTT) cell assay and Annexin V/propidium iodide (PI) staining.

Results

Here we are reporting the inhibitory effect of BHB on insulin glycation and generation of insulin-AGE as a possible explanation for insulin resistance. Moreover, the protective effect of BHB on consequential glycation derived liposomal lipid peroxidation as a causative event in microglial apoptosis is reported.

Conclusion

The reduced insulin fibril formation, structural inertia to glycation involved conformational changes, anti-lipid peroxidation effect, and increasing microglia viability indicated the protective effect of BHB that disclose insight on the possible preventive effect of BHB on Alzheimer’s disease.     

The cardiac atria are chambers of active remodeling and dynamic collagen turnover during evolving heart failure

OBJECTIVES: The role of atrial myocytes and extracellular matrix (ECM) changes in atrial chamber remodeling was studied in a canine model of heart failure (HF). BACKGROUND: Cardiac remodeling is a key process mediating the progression of HF. Studies of the structural mechanisms of cardiac remodeling have been limited to the left ventricle. The structural alterations associated with atrial chamber remodeling in evolving HF have not been studied. METHODS: Age- and weight-matched dogs were subjected to right ventricular pacing (240 beats/min) for one and three weeks to produce early and severe HF, respectively. Atrial tissues were assessed for myocyte and ECM changes. RESULTS: Right atrial and left atrial (LA) pressures were significantly increased in early and severe HF. The LA wall tension index was significantly increased at both HF stages by 116% and 443%, respectively. Atrial collagen synthesis and degradation were significantly increased in severe HF. Gelatinase activity was significantly increased at both early and severe stages of HF. Gelatin zymography showed increased matrix metalloproteinases (MMP)-9 with early HF and increased MMP-2 with severe HF. The LA wall tension index was significantly correlated with gelatinase activity and collagen synthesis. Although total atrial collagen content was not changed, disarray of collagen fibers was observed. Atrial myocyte hypertrophy without evidence of apoptosis was also present in severe HF. CONCLUSIONS: There is marked atrial chamber remodeling in canine pacing-induced HF, which is characterized by myocyte hypertrophy and dynamic collagen turnover. Atrial remodeling may contribute to the development of atrial arrhythmias and pulmonary hypertension and could offer a novel therapeutic target.     

DNA Methylation Profiling Revealed Promoter Hypermethylation-induced Silencing of p16, DDAH2 and DUSP1 in Primary Oral Squamous Cell Carcinoma

Background: Hypermethylation in promoter regions of genes might lead to altered gene functions and result in malignant cellular transformation. Thus, biomarker identification for hypermethylated genes would be very useful for early diagnosis, prognosis, and therapeutic treatment of oral squamous cell carcinoma (OSCC). The objectives of this study were to screen and validate differentially hypermethylated genes in OSCC and correlate the hypermethylation-induced genes with demographic, clinocopathological characteristics and survival rate of OSCC.Methods: DNA methylation profiling was utilized to screen the differentially hypermethylated genes in OSCC. Three selected differentially-hypermethylated genes of p16, DDAH2 and DUSP1 were further validated for methylation status and protein expression. The correlation between demographic, clinicopathological characteristics, and survival rate of OSCC patients with hypermethylation of p16, DDAH2 and DUSP1 genes were analysed in the study.Results: Methylation profiling demonstrated 33 promoter hypermethylated genes in OSCC. The differentially-hypermethylated genes of p16, DDAH2 and DUSP1 revealed positivity of 78%, 80% and 88% in methylation-specific polymerase chain reaction and 24% and 22% of immunoreactivity in DDAH2 and DUSP1 genes, respectively. Promoter hypermethylation of p16 gene was found significantly associated with tumour site of buccal, gum, tongue and lip (P=0.001). In addition, DDAH2 methylation level was correlated significantly with patients'' age (P=0.050). In this study, overall five-year survival rate was 38.1% for OSCC patients and was influenced by sex difference.Conclusions: The study has identified 33 promoter hypermethylated genes that were significantly silenced in OSCC, which might be involved in an important mechanism in oral carcinogenesis. Our approaches revealed signature candidates of differentially hypermethylated genes of DDAH2 and DUSP1 which can be further developed as potential biomarkers for OSCC as diagnostic, prognostic and therapeutic targets in the future.     

Selective photocatalytic oxidation of aromatic alcohols to aldehydes with air by magnetic WO3ZnO/Fe3O4. In situ photochemical synthesis of 2-substituted benzimidazoles

Recently, visible light-driven organic photochemical synthesis has been a pioneering field of interest from academic and industrial associations due to its unique features of green and sustainable chemistry. Herein, WO₃ZnO/Fe₃O₄ was synthesized, characterized, and used as an efficient magnetic photocatalyst in the preparation of a range of 2-substituted benzimidazoles via the condensation of benzyl alcohol and o-phenylenediamine in ethanol at room temperature for the first time. The key feature of this work is focused on the in situ photocatalytic oxidation of benzyl alcohols to benzaldehydes under atmospheric air and in the absence of any further oxidant. This new heterogeneous nanophotocatalyst was characterized via XRD, FT-IR, VSM and SEM. Short reaction time, cost-effectiveness, broad substrate scope, easy work-up by an external magnet, and excellent product yield are the major advantages of the present methodology. A number of effective experimental parameters were also fully investigated to clear broadness and generality of the protocol.

WO₃ZnO/Fe₃O₄ is used as a magnetic photocatalyst in the preparation of 2-substituted benzimidazoles in EtOH at RT. The key feature is the in situ photocatalytic oxidation of benzyl alcohols to benzaldehydes under atmospheric air in the absence of further oxidant.     

Persistence of hepatitis B vaccine immunity in hemodialysis patients

Although the efficacy of hepatitis B vaccines in patients undergoing chronic hemodialysis (HD) treatment has been documented, the persistence of immunity in this population remains largely unknown. In this study we evaluated the persistence of hepatitis B vaccine immunity in HD patients. We followed 37 hepatitis B vaccinated HD patients (following a four-dose vaccination schedule of 40 mug injections intramuscularly in the deltoid muscle at 0, 1, 2, and 6 months) for up to one year to evaluate the persistence of immunity (as indicated by serum levels of hepatitis B surface antibody (anti-HBs) equal to or higher than 10 IU/L). One year after vaccination, 18.9% of patients had lost their anti-HBs (transient responders), while 81.1% still had detectable antibodies in the serum (persistent responders). From 81.1% of persistent responders 11.5% and 88.5% were weak and high responders, respectively. There was no significant difference between persistent and transient responders regarding age, sex, or nutritional factors. We did not find any factors that related to maintaining protective levels of anti-HBs in HD patients. It seems that an antibody titer above 100 IU/L following vaccination is necessary in order to maintain that level of antibody one year later.