Caveolin 3 gene and mitochondrial tRNA methionin gene in Duchenne muscular dystrophy

It was recently reported that Duchene muscular dystrophy [DMD] patients and mdx mice have elevated levels of caveolin-3 expression in their skeletal muscles. However, it remains unknown whether this increased caveolin-3 levels contribute to the pathogenesis of DMD. Also mitochondrial DNA mutation in the tRNA methionin [tRNA Met] gene has been shown to be associated with muscle weakness, severe exercise intolerance, lactic acidosis and growth retardation. Since DMD is X-linked maternally inherited disease, mitochondrial mutation in tRNA [Met] gene can be suspected to be the cause for the inefficient splicing of dystrophin gene during its expression and can be implicated as the cause of dystrophin inactive protein. The aim of the present study is to investigate whether mutations in caveolin gene leads to its increased expression and/or mutation in the tRNA [Met] gene can be associated with DMD pathogenesis. Expression of caveolin mRNA by RT-PCR and mutations in caveolin gene and tRNA [Met] gene were measured in 28 patients presented with DMD symptoms using the single strand conformation polymorphism assay [SSCP]. Results gave further proof to decreased expression of inducible nitric oxide synthase [iNOS] mRNA, which leads to increased expression in caveolin 3 mRNA in lymphocytes of DMD patients compared to controls. However using SSCP, there was no evidence for tRNA [Met] gene mutation among DMD patients and only one patient presented a mutation in the caveolin gene compared to controls. There is an inverse relation between iNOS and Caveolin 3 in lymphocytes of DMD patients compared to controls. However, Caveolin 3 gene mutation is excluded as the main cause of increased caveolin gene expression. Also, there was no evidence for tRNA [Met] gene mutation among DMD patients

Individual susceptibility to vernal keratoconjunctivitis and ameliorative effects of antioxidants on its symptoms

An allergic response is associated by an increase in the expression and activity of the inducible nitric oxide synthase [iNOS] an inducible enzyme present endothelial cells and polymorphonuclear leucocytes [PMN] and moncytes. Vernal keratoconjunctivitis [VKC], is a recurrent or chronic ocular allergic eye disease affecting young individual living in a hot climate. This study was designed to test a hypothesis that the immune changes seen in Vernal keratoconjunctivitis during the summer or spring season is a function of the effect of hot weather on the of polymorphonuclear leucocytes [PMN] and monocytes in susceptible individuals, by evaluating the role of nitric oxide [NO] and NO-synthase [iNOS] in the pathogenesis of allergic conjunctivitis and the effect of supplementing those patients with antioxidants in order to ameliorate ocular allergic effects. 40 patients with typical VKC were divided into two groups. Antioxidants were given as a supplement to one group [n=20], while the second group did not [n=20]. Both groups were followed up periodically for clinical picture and status and were compared with a third healthy age and socio-economic matching control group. Blood samples were obtained before treatment, after one month and at the end of the whole follow up period. Blood markers of oxidative stress including: protein carbonyls, nitrite. DNA fragmentation and apoptosis in circulating lymphocytes of VKC before and after antioxidant therapy during the severity of the disease were evaluated. Also neutrophil's individual ability to express iNOS was assessed. Vernal Keratoconjunctivitis patients exhibited increased individual ability in the production and expression of nitric oxide by their stimulated neutrophils [4.5 +/- 0.05 nmol/ 10[5] cell vs. 2.5 +/- 0.09 nmol/ 10[5] cell] compared to controls. Both biochemical and clinical picture proved that a supplement of antioxidants to the traditional topical treatment would significantly improve the patients ocular allergic symptoms even after seizure of topical treatment, while recurrence of symptoms and signs occurred in group two after stoppage of the topical steroidal treatment. The antioxidant supplemented group exhibited decreased markers of oxidative stress at the end of the first month follow-up period compared to those patients that were not supplemented with antioxidants: Protein carbonyls [0.57 +/- 0.34 vs. 0.93 +/- 0.05]; Percentage of DNA fragmentation per total DNA in combined plasma and leucocytes after one month [0.7% +/- 0.09 vs. 0.93 +/- 0.05] and plasma nitrite after one month [2.4 +/- 0.5 vs. 3 +/- 0.19 nMol/ml] in VKC patients compared to controls respectively. The associated immunological and inflammatory systemic condition of VKC recommends the use of antioxidant mixture in order to emeliorate the ocular allergic effects associated with VKC. Antioxidant use proved its efficiency and prevent recurrence of VKC even after stoppage of the steroid. Also individuals with increased neutrophil's activity have higher individual susceptibility to develop VKC

Effects of high and low doses of He:Ne laser irradiation on human lymphocytes in vitro

Low level laser irradiation [LLLI] can cause cell proliferation, differentiation, or death; however, the cellular mechanisms of these effects of LLLI, at high or low fluences. The exact mechanism of action of the laser radiation with living cells is not yet understood. Since He:Ne has been shown to increase mitochondrial respiration it is relevant to study the effect of different high and low laser fluencies on mitochondrial proteins involved in apoptosis. The aim of this work was to study safety aspects of the effect of different high and low He:Ne laser fluencies on mitochondrial proteins involved in apoptosis. Expression of Bax mRNA and Bcl-2 lymphocytes together with mitotic index and viability percentage together with the frequency of micronuclei were evaluated after exposing cells with a 10 mW He-Ne laser [wavelength 632.8 nm] at energy densities of 1, 2.5, 5, 10, 35 and 60 J/cm[2]. At low level He:Ne laser irradiation induced a significant increase in Bcl-2 and an increase in viable cell percentage. However, at moderate dose there was an increase in Bax mRNA expression and decrease in viability percentage at doses ranging from 15 -60 J /cm[2]. Bcl-2 family proteins are stimulated by He:Ne laser irradiation, which is safe at doses below 10 J /cm[2] and comparatively very safe, when compared with low doses of ionizing radiation

Effect of He: Ne laser on neutrophils phagocytic activity

The aim of this study is to investigate the effects exerted by He:Ne laser [632.8 nm] low level laser irradiation [LLLI] on neutrophil activity in response to op sonized zymosan. Human buffy coat leukocytes primed with opsonized zymosan were exposed to 10 mw He:Ne laser at energy densities of 1-5 J/cm[2]. Phagocytic activity was assessed by measuring percent of phagocytosis, H2O2 and nitric oxide [NO] generation together with neutrophil-associated nitric oxide synthetase activity [NOS] and Arginase I mRNA. LLLI stimulated neutrophil's phagocytic activity and free radicals generation. This effect was intensified, when neutrophils were dually stimulated by LLLI and opsonized zymosan within the dose range of 3-5 with maximum activity at 5 J/cm[2]

He: Ne laser irradiation induced survival and cell cycle progression effects on human circulating mononuclear cells in vitro

The biostimulation and therapeutic effects of low-power laser radiation of different wavelengths and light doses are well known, but the exact mechanism of action of the laser radiation with living cells is not yet understood. The aim of the present study was to investigate whether He:Ne laser irradiation induced mitogenic stimulation on human blood circulating mononuclear cells in vitro displays peculiar features of cell cycle regulation. Buffy-coat human leukocytes were irradiated with He-Ne [10 mW] at energy densities of 1, 2.5 and 5 J/cm[2] and cultured in medium 199 without any supplements in the absence and presence of phytohaemagglutinin for120 hours. Detailed analysis of viability percentage, telomerase activity and gene p53 mRNA expression was performed and compared with a similar analysis of phytohaemagglutinin stimulated and non-stimulated mononuclear cells. Results showed that laser induced telomerase activity in blood mononuclear cells throughout the five consecutive days post laser irradiation, reaching its highest level at 72 hours post laser irradiation and was significantly higher in PHA stimulated cells compared to laser irradiated cells, where 5 J/cm[2] displayed the highest activity. There were no changes in gene p53 mRNA expression at zero compared to 72 hours post laser irradiation. However, it was non-significantly higher in PHA stimulated cells compared to laser stimulated cells. Results threw some light over the mechanism encountered by lymphocytes in the process of He-Ne laser induced biostimulation

Non small cell lung cancer molecular biomarkers in blood

Several studies have reported significant correlations between individual telomerase activity and apoptosis in malignant tumours including lung cancer. Such studies were carried out on tissue biopsies from the malignant tissue. The aim of the present study was to determine molecular biological parameters that can he used for early biological predisposition for lung cancer. Telomerase activity and Bcl-2 anti-apoptotic protein in circulating lymphocytes, plasma nitric oxide. epidermal growth factor and epidermal growth 'actor receptor were measured in the blood of 25 non small cell lung cancer [NSCLC] patients amid in 20 normal age and socio-economic matching controls, Results revealed significant increase in telomerase activity [53 +/- 8.2 vs. 19.5 +/- 4.8. p<0.0001] between cancer patients and normal controls, significant lower levels of Bcl-2 [7 2 +/- 1,5 vs. 10.4 +/- 1.4 micro g/ml, t 7.3. p<0.0000001] among cancer patients compared to normal controls, However, there were significantly higher levels of epidermal growth factors [6.2 +/- 2 05 vs 0.2 +/- 0.01 pg/ml], epidermal growth factor receptor EGFR [134 +/- 4.7 vs. 102 +/- 2.2 fmol/ml] and plasma nitrate/nitrite [18.8 +/- 4.7 vs. 12.5 +/- 5 micro g/ml. 1=-5.25. p<0.0001] in lung cancer patients compared to controls. This study reveals that Bcl-2 levels in circulating peripheral blood are weak biomarkers for lung cancer, while increase in plasma telomerase activity, NO and EGF levels can he used as reliable biomarkers for cancer prognosis

Markers of oxidative stress in nurses exposed to glutaraldehyde in gynaecologic endoscopy unit

Objectives: to investigate oxidative stress of glutaraldehyde of occupationally exposed nurses

Subjects and Methods: in 20 controls and 20 nurses involved in sterilizing endoscopy at Gynecological Departments, plasma levels of nitrite/nitrate, malondialdehyde, the frequency of chromosomal aberrations and DNA-protein crosslinking in circulating lymphocytes together with the expression of inducible nitric oxide synthase [iNOS] were investigated

Results: there was a significant increase in the level of plasma malondialdehyde in the "glutaraldehyde" exposed group [mean 13.9 +/- 4 versus 6.11 +/- 1.8 nmol/ml] when compared with controls. A concomitant increase in the percent of all types of chromosomal aberrations, including dicentrics, rings, gaps, acentrics, chromosome and chromatid breaks among glutaraldehyde exposed nurses when compared with the controls. DNA- protein crosslinks showed a significant increase in these women [20-39%] when compared with controls [16-28%]. Both plasma nitrite/nitrate level and iNOS expression showed significant increase in exposed nurses [4.9 +/- 1.5 vs. 1.5 +/- 0.6 nmol/ml, p<0.001] for nitrite/nitrate and increased expression in iNOS in neutrophils of glutaraldehyde in exposed nurses compared to controls

Conclusion: Results indicate the genotoxic effect of "glutaraldehyde" among nurses working at endoscopy units and call for adequate preventive measures to protect such women

Measurement of markers of apobtosis and oxidative stress in maternal blood, cord blood and amniotic fluid in cases of premature rupture of membranes

Objective: to determine markers of apoptosis and oxidative stress in maternal blood, cord blood and amniotic fluid in cases of premature rupture of membrane at labor [PROM], preterm premature rupture of membranes [P-PROM] compared with laboring women with intact membranes

Subjects and Methods: antiapoptotic gene Bcl-2, nitrite/nitrate and lipid peroxidation [malondialdehyde MDA] were determined in amniochorion, maternal and cord leucocytes, plasma and amniotic fluid from gestational age-matched groups of women at labor. Samples were collected from the three groups of women [PROM, P-PROM and women with intact membrane as control group]

Results: maternal blood leucocytes and amniotic fluid Bcl-2 protein levels showed no significant differences between PROM and controls. However, there was a significant decrease of cord blood leucocytes Bcl-2 in cases of PROM and P-PROM. The decrease was more pronounced in the latter when compared to the former. Maternal, cord plasma and amniotic fluid MDA levels were significantly elevated in PROM and P-PROM compared to control values. The mean fold rise were 1.2, 1.9% respectively for maternal plasma, 1.2, 1.7% respectively for cord plasma and 2.5% for amniotic fluid. Matemal, cord plasma and amniotic fluid nitrite/nitrate levels were significantly elevated in PROM and P-PROM compared to control values. The mean fold rise were 1.3, 1.6% for maternal plasma and 1.2, 1.3% for cord plasma and 1.44% for amniotic fluid, respectively. Apoptotic cells was positively correlated with nitrite/nitrate while negative correlations, existed between Bcl-2 and nitrite/nitrate, MDA and apoptotic cells

Conclusion: enhanced oxidative stress [increased MDA], inflammation process [increased NO] and decreased antiapoptotic Bcl-2 are the characteristic features associated with P-PROM and PROM and may play causative role by affecting membrane integrity