Assessing extent of single stranded DNA damage in oral mucosal cells of patients with oral squamous cell carcinoma and its correlation with TNM staging.

Context: This study was carried out on the assumption that oral mucosal cells might show DNA damage in oral squamous cell carcinoma (OSCC). Aims: To evaluate the extent of DNA damage in oral smears of patients with OSCC and determine correlation if any of the extent of DNA damage to TNM staging of oral cancer. Settings and design: A randomized controlled study at a regional cancer centre was designed for this project. Smears were taken from lesion proper of 30 patients with OSCC and from the buccal mucosa of 30 normal healthy volunteers. Materials and methods: Collected cells were centrifuged and single-cell gel electrophoresis (SCGE) assay was performed. DNA damage was visualized under a fluorescent microscope. Statistical analysis used : Mean DNA damage levels of both the groups were measured and statistically analyzed with students' test. The extent of DNA damage was correlated with the TNM stages by employing the one way ANOVA 'F' technique. Results: High statistical significance (P < 0.0001) was found in DNA damage levels between control and study groups. A stepwise increase in DNA damage levels with high statistical significance (P < 0.005) was also found between all the TNM stages. Conclusions: Statistically significant increased DNA damage levels in OSCC patients and their correlation to clinical staging suggest that comet assay may be used effectively to assess the prognosis of OSCC.

Prevalence of methylenetetrahydrofolate reductase C677T and A1298C polymorphisms in Egyptian patients with type 2 diabetes mellitus

Type 2 diabetes mellitus [T2DM] is a major public health problem around the world. The C677T and A1298C polymorphisms of the methylenetetrahydrofolate reductase [MTHFR] gene have been reported to be associated with T2DM and its complications. This study is a case-control study which was performed to clarify the association between polymorphisms in these two genes and T2DM among Egyptians. Study population [n = 120] consists of 60 Egyptian diabetic patients and 60 healthy controls. The MTHFR C677T and A1298C polymorphisms were gen-otyped by polymerase chain reaction, followed by enzymatic digestion with Hinfl and MboII enzymes, respectively. C677T and A1298C genetic polymorphisms conveyed an increase in T2DM risk [OR = 3.5, 95% CI = 1.1-11.6, p = 0.032 and OR = 2.2, 95% CI = 0.7-6.9, p = 0.004 respectively] Additionally, no significant associations between lipid/glucose metabolic indexes with MTHFR genotypes among diabetic patients were observed. Combined MTHFR gene polymorphisms revealed higher T2DM risk in homozygous and heterozygous forms compared to single gene polymorphism with pronounced risk in C677T/CT-A1298C/CC combined form [OR = 6.56, 95% CI = 0.76-56.2, p 0.041]. In conclusion, our data suggest that MTHFR C677T and A1298C polymorphisms are risk factor for T2DM in Egyptian patients. Also, the two gene polymorphisms may act synergistically to increase the risk of diabetes. Furthermore, it should be noted that the size of the studied population was relatively small and therefore, large-scale prospective studies are needed to confirm these findings

Técnicas electroforéticas en el HYDRASYS 2. Utilidad diagnóstica en diferentes enfermedades / Electrophoretic techniques in HYDRASYS 2. Diagnostic usability in different diseases

Se presentan los resultados de la estandarización de las técnicas de electroforesis de hemoglobina (Hb), isoenzimas de la deshidrogenasa láctica (LDH) y proteinuria en el equipo Hydrasys 2, así como el estudio de pacientes atendidos en el Instituto de Hematología e Inmunología y en otros centros hospitalarios del país. Se realizó el diagnóstico de 149 portadores de hemoglobinopatías (AS, AC, b talasemia heterocigótica, variante rápida), 60 enfermos (SS, SC, CC), 24 pacientes con a talasemia o deficiencia de hierro y se cuantificó la hemoglobina fetal a 93 casos con hemoglobinopatía S. Se determinaron los valores normales de actividad e isoenzimas de LDH en la población mediante el estudio de 50 donantes de sangre. En los pacientes con anemia drepanocítica se encontró un aumento significativo de la isoenzima 1 (p= 0,000) y disminución de isoenzimas 3 (p= 0,002). Se realizó el estudio de proteínas en orina a 8 pacientes con enfermedades hematológicas que presentaron microalbuminuria al menos en 2 ocasiones, con concentraciones ³ 0,04 g/L. En 2 pacientes el resultado fue normal; en 2 se encontraron proteínas de origen tubular; y en otros 2, proteínas de origen glomerular

We present the results of the standardization of the techniques of electrophoresis of hemoglobin (Hb), lactate dehydrogenase isoenzymes (LDH) and proteinuria in HYDRASYS 2 equipment, and the study of patients treated at the Institute of Hematology and Immunology and other hospitals in the country. 149 hemoglobinopathies carriers were diagnosed (AS, AC, b thalassemia heterozygous fast variant), 60 patients (SS, SC, CC), 24 patients with athalassemia or iron deficiency. Fetal hemoglobin was quantified in 93 cases with hemoglobinopaty S. Normal values of activity and LDH isoenzymes were determined in the population through the study of 50 blood donors. In patients with sickle cell anemia we found a significant increase in isoenzyme 1 (p=0.000) and isozyme 3 decreased (p=0.002). We performed the study of proteins in urine in 8 patients with hematologic malignancies who had microalbuminuria at least 2 times, with concentrations ³ 0.04 g / L. In 2 patients the results were normal, in 2 proteins were tubular origin, and in 2, proteins of glomerular origin

Genotoxicity of cadmium in rat lung cells assessed by an alkaline comet assay and the possible protective role of selenium

Cadmium is a highly toxic heavy metal with many hazardous effects. Selenium is an essential trace element with antioxidant properties. To evaluate the cadmium-induced DNA damage in rat lung cells and the possible protective role of selenium. Thirty-six adult male albino rats were used in this experiment. They were divided into four equal groups. Group I was the control group. Group II included rates that were injected intraperitoneally with cadmium chloride [CdCl[2]] [1.0 mg/kg/day] for 8 weeks. Group III included rates injected intraperitoneally with sodium selenite [Na[2]SeO[3]] [0.25 mg/kg/day] for 8 weeks. Group IV included rats injected intraperitoneally with both CdCl[2] [1.0 mg/kg/day] and Na[2]SeO[3] [0.25 mg/kg/day] for 8 weeks. At the end of the experiment, the lungs of rats were taken and processed for single-cell gel electrophoresis [comet assay]. In the control group, comets appeared with large bright heads and small tails. The injection of CdCl[2] in group II induced DNA damage in rat lung cells, indicated by increased migration in the comet assay. Comet cells appeared with small heads and long tails. Statistically, there was a significant increase in tail length, tail moment, and tail% DNA compared with the control. Injection of selenium alone did not show any difference from the control. Concomitant administration of both cadmium and selenium resulted in a partial decrease in the DNA damage in rat lung cells indicated by decrease in DNA migration in the comet assay. Comet cells appeared with large heads and relatively small tails compared with those of group II. A significant decrease was also observed in tail length, tail moment, and tail% DNA compared with group II. CdCl[2] could significantly induce DNA damage in rat lung cells. It was suggested that selenium could partially ameliorate DNA damage induced by cadmium

Using some recent techniques in diagnosis of some zoonotic bacterial diseases transmitted through milk

A total of 200 different samples [100 raw milk, 50 Kareish cheese and 50 yogurts] were collected from dairy shops, street venders and supermarkets from different localities in Alexandria Province to be investigated for presence of bacterial pathogens and to evaluate plasmid profiling as a tool for detection of bacteria transmitted through milk. The results of bacteriological examination firstly revealed that the rates of isolation of E. coli were 18, 26 and 18% from raw milk, kariesh cheese and yoghurt samples, respectively; secondly, results revealed that the rates of isolation of Salmonella species were 5, 6 and 2% from raw mill, kariesh cheese and yoghurt samples, respectively and finally they clarified that the rates of isolation of Staph. aureus were 44, 34 and 18% from raw milk, kariesh cheese and yoghurt samples, respectively. Serological identification of E. coli and Salmonella spp. was carried out. The public health significance of isolated bacteria and the diagnostic value of plasmid were discussed

Análise da composição do matriz extracelular das camadas urotelial e muscular da bexiga de mulheres em diferentes idades / Analysis of the extracellular matriz of urothelial and muscle layers of the bladder in women at different ages

Acomplacência da bexiga depende de músculos lisos, fibras colágenas, fibras elásticas e suas relações. O objetivo deste trabalho é determinar a composição da matriz extracelular em amostras de bexigas normais através de análise bioquímica de colágeno e glicosaminoglicanos em amostras obtidas de mulheres em diferentes grupos de idade, analisando separadamente as camadas urotelial e muscular. Avaliamos 17 amostras de bexiga divididas em três grupos: infância (N=5), menacme (N=6) e pós-menopausa (N=6). As bexigas foram analisadas para concentração de GAG total e colágeno e para análise qualitativa de GAG por eletroforese em gel de agarose. Na camada muscular, não houve diferença entre os grupos tanto para GAG quanto para colágeno. Na camada urotelial, a análise da concentração de colágeno não mostrou diferença entre os grupos, mas a concentração de GAG no grupo da pós-menopausa (0.21 +- 0.12 ug de ácido hexurônico/mg de tecido seco) apresentou diferença em relação aos grupos do menacme (1.78 +- 1.62 ug de ácido hexurônico/mg de tecido seco) e da infância (2.29 +- 1.32 ug de ácido hexurônico/mg de tecido seco). Nosso trabalho concluiu que a concentração de GAG está substancialmente diminuída na cadama urotelial da bexiga de mulheres na pós-menopausa.

Bladder compliance is dependent on smooth muscle, collagen fibers, elastic fiber and their ratios. The luminal surface of the urothelium is covered by an adhering glycosaminoglycan (GAG) layer. The aim of this study was to determine the composition of the extracellular matrix (ECM) in normal samples of women bladders through biochemistry analysis of collagen and GAG on samples obtained from individuals from different age groups, analyzing separately the urothelial and muscular layers. We studied samples taken from bladders of 17 patients divided in three different groups: childhood (N=5), menacme (N=6) and menopause (N=6). Bladders were analyzed for total GAG and collagen concentration per mg dry tissue and for the contents of GAG species, as determined by agarose electrophoresis and reported as the percent of total sulfated GAG. In muscular layer, collagen and GAG concentration showed no difference between groups. In urothelial layer, collagen concentration showed no difference between groups but GAG concentration in menopause (0.21 +- 0.12 ug hexuronic acid/mg dry tissue) was different from menacme (1.78 +- 1.62 ug hexuronic acid/mg dry tissue) and childhood (2.29 +- 1.32 ug hexuronic acid/mg dry tissue). There was no difference between sulfated GAG in three groups. In conclusion, GAG concentration in urothelial layer was substantially lower in menopause women.

Proteomic profiles characteristic of Schistosoma mansoni infection in patients from two different endemic areas

Knowledge of the Schistosoma proteome may greatly enhance our understanding of its physiological and pathological processes at the molecular level and may provide new models for diagnosis and development of vaccines or drugs. Despite the importance of this approach, Schistosoma proteomic research in Egypt and Brazil is still incipient. To identify the profiles of S. mansoni serum reactive proteins [SRP] that may characterise clinically relevant subgroups of patients with schistosomiasis from two different endemic regions; Egypt and Brazil. Soluble egg antigen [SEA] and soluble worm antigen preparation [SWAP] of Egyptian and Brazilian strains of S. mansoni were resolved by two-dimensional gel electrophoresis [2-DE]. Serum samples were collected from patients with intestinal [INT] and hepatosplenic [HS] schistosomiasis from both countries. Sera were probed using 2-dimensional western blot [2-DWB] against the corresponding separated antigens proteins [i.e. Egyptian SEA and SWAP were probed separately against Egyptian sera, while Brazilian SEA and SWAP were probed separately against Brazilian sera] to determine the isoelectric focusing point [pI] and molecular weight [MW] of the SRP. Both Egyptian and Brazilian strains antigens gave similar electrophoretic patterns in their p1 amid MW, where 97 soluble proteins from SEA and 125 from SWAP were resolved by 2-DE. For SEA, 11 and 12 proteins uniquely reacted to sera of INT and US patients respectively, and 62 proteins reacted to sera from patients with concurrent INT and HS schistosomiasis. Out of 85 SEA reactive proteins; 16 were reactive to >/= 50% of sera of patients with the INT or HS clinical form of the disease. Regarding SWAP, 24 and 10 proteins uniquely reacted to sera of INT and HS patients respectively, and 35 proteins reacted to sera from patients with both clinical forms of schistosomiasis. Out of 69 SWAP reactive proteins, 6 proteins were reactive to >/= 50% of sera from Egyptian patients with INT schistosomiasis. Sera of patients from Egyptian and Brazilian populations exhibited significant differences [P=0.032] for recognition of the immune reactive proteins to SEA and SWAP in the two different clinical forms of the diseases. Further definition of reactive proteins could provide novel targets for vaccine design and therapeutic intervention. Further detailed studies on large population scale are recommended to correlate different proteomic data among different clinical forms of schistosomiasis in an attempt to generate a vaccine valuable in schistosomiasis control in endemic areas

Serum lipoproteins in liver diseases.

Agarose gel electrophoresis is a rapid technique yielding dependable results than any other conventional older ones, more cases in shorter time can be conveniently studied using this technique. In cirrhosis and hepatitis patients the alpha lipoprotein and the pre beta lipoprotein are found to be lower than that of normal healthy subjects.

Evaluation of a Multiplex PCR Kit Used for Detecting Y Chromosome Microdeletions / 대한진단검사의학회지

BACKGROUND: In addition to Klinefelter's syndrome, microdeletion of Yq is the most common genetic cause of male infertility; 15% of azoospermic or 5-10% of oligozoospermic males have Yq deletions. We evaluated a Yq microdeletion kit (LG Life Sciences, Korea) for identifying microdeletions in the azoospermic factor (AZF) regions of the Yq. METHODS: The kit was designed to amplify 3 regions of the AZF gene (AZFa, AZFb, and AZFc) using 15 sequence-tagged sites. We evaluated the preclinical performance of the kit. For clinical validation, 58 patients including 25 idiopathic azoospermic or oligozoospermic patients were examined. RESULTS: We observed clear bands on electrophoresis of DNA, up to a DNA concentration of 3.12 ng/microliter; the known microdeletion regions of all 6 reference cell-lines (Coriell, USA) were accurately detected and no false positive/negative results showed with normal female (n=11) and fertile male (n=15) specimens. This kit could identify the same microdeletions in the common regions, similar to another commercial kit. Among the 58 male infertile patients, 7 (12.1%) had microdeletions of the Yq. Among the idiopathic azoospermic (n=22) and oligozoospermic (n=3) patients, 3 (12.0%) had microdeletions. Further, 2 of 21 varicocele patients (9.5%), 1 of 4 patients with testicular failure, and 1 patient with a 45,X/46,XY mosaic had microdeletions. CONCLUSIONS: The kit was effective for detecting microdeletions of the Yq. We identified microdeletions in 12% of the infertile patients. This Y chromosome microdeletion detection kit is useful for screening Yq microdeletions in infertile patients.

Toxic effects of acrylamide on the prostate of adult male albino rats and the role of pretreatment with vitamin C: immunohistochemical and DNA electrophoresis study

Acrylamide [AA] is a widely studied industrial chemical that is neurotoxic and mutagenic to somatic and germ cells. A considerable public concern about cancer risk from acrylamide-rich foods followed the announcement that high concentrations of acrylamide are found in starch-containing foods cooked at high temperatures. Data concerning the toxic effect and DNA damage of acrylamide in the prostate are still scarce, so the aim of this work tries to prove the toxic effect of acrylamide on the prostate by using histological and immunohistochemical studies, and to determine its potential mutagenic effect on DNA of the prostatic cells. The possible antimutagenic effect of ascorbic acid is also evaluated in this study. The experimental study was conducted on sixty adult male albino rats. Twenty adult control male albino rats [group I] subdivided into two subgroups, a and b, received distilled water and vitamin C orally. The experimental group [group II] is divided equally into four subgroups: a, b, c, and d. Each group consists of ten rats, receiving acrylamide in a dose of 30 mg/Kg/day and 60 mg/Kg/day in subgroup II [a] and subgroup II [b] respectively. Subgroup II [c] and Subgroup II [d] received vitamin C in dose of 100 mg/kg /day one hour prior to acrylamide ingestion in a dose of 30 mg/Kg/day and 60 mg/Kg/day respectively for four weeks The histological picture of the prostate in experimental subgroup II a revealed signs of dysplasia in the form of nuclear pleomorphism, loss of nuclear polarity, enlarged nucleus with evident nucleolus and discontinuity of the basal cell layer. Some acini showed hyperplasia of its epithelial lining with apparent increase in stromal thickness. Rats in subgroup II b were presented with post atrophic hyperplasia in the form of atrophy and thinning of the epithelial lining of some acini. Focal areas of acinar hyperplasia with discontinuity of the basal cell layer were present. On pretreatment with vitamin C, the prostate showed apparent improvement of the epithelial cells height in subgroups II c and d. Immunohistochemical stained sections of acrylamide treated groups revealed estrogen receptor alpha [ER alpha] expressed as heavily brown dots in the prostatic stroma and in many nuclei of the acinar epithelial cells .The groups treated with vitamin C prior to acrylamide showed a slight decreased in ER alpha expression than groups treated with acrylamide alone. A significant increase in DNA damage was present in rats treated with 30 mg/kg/day acrylamide. Increasing the dose of acrylamide to 60 mg/kg/day led to significant increase in DNA damage of prostatic cells. Administration of vitamin C prior to acrylamide resulted in a significant decrease in DNA damage. It is concluded that acrylamide toxicity in adult male albino rat prostate [which was clarified by histological, immunohistochemical studies and DNA electrophoresis] had a direct dose response relationship and its association with acinar dysplasia serves as a warning that concurrent malignancy may exist. Vitamin C proved to have a protective role when received before exposure to acrylamide

Characterization of cellulolytic activities of Bjerkandera adusta and Pycnoporus sanguineus on solid wheat straw medium / Caracterización de las actividades celulolíticas de Bjerkandera adusta y Pycnoporus sanguineus en un medio sólido de paja de trigo

Cellulolytic properties of two white rot fungi, Bjerkandera adusta and Pycnoporus sanguineus, cultivated on wheat straw agar medium, were characterized and compared. Optimal growing parameters for maximum enzyme production for both fungi were wheat straw medium pH 5 and 28ºC. B. adusta showed, on the 6th day of culture, carboxymethylcellulose (CMC)ase activity levels 1.6 times higher than maximal P. sanguineus activity, achieved on the 8th day. B. adusta supernatants also displayed higher activity levels towards xylan (3.6-fold) compared to those of P. sanguineus. However, enzymes from P. sanguineus were more robust resisting one hour incubation at high temperatures (up to 80ºC), and exhibiting activity and stability in pH range from 2 to 8. Cellulolytic activities, with molecular masses ranging from 25 to 90 kDa, from the two species were detected in zymograms.

Perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel) criadas em cativeiro / Serum protein electrophoresis profile of the rattlesnake Crotalus durissus terrificus kept in captivity

As serpentes peçonhentas dos gêneros Bothrops e Crotalus têm sido mantidas em cativeiro visando à extração de venenos para a produção de imunobiológicos. O conhecimento da fisiologia desses animais e as alterações na concentração de proteínas e suas frações séricas são importantes para a identificação precoce de importantes enfermidades que cursam com estados de hipoproteinemia e hiperproteinemia. O objetivo do trabalho foi determinar a concentração de proteína total e o perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel) criadas em cativeiro. Foram colhidas amostras de sangue da veia coccígea ventral de 21 serpentes adultas e sadias, divididas em dois grupos: Grupo 1 de 12 machos com peso médio de 588,89±193,55g, e Grupo 2 de nove fêmeas com peso médio de 708,33±194,04g. A proteína total sérica foi determinada pelo método de refratometria e a eletroforese em gel de agarose. Obtiveram-se valores da proteína total sérica (g/dL) de 4,51±0,50 para machos e de 4,82±0,72 para fêmeas, e para machos e fêmeas de 4,64±0,61. Foram identificadas pela eletroforese quatro frações protéicas (g/dL): albumina, a, b, g-globulinas e calculada a relação albumina:globulina. As serpentes fêmeas apresentaram maiores valores para as variáveis, albumina e para a relação albumina/globulina (AG) diferindo significativamente (P<0,05) do grupo de machos, porém sem significado clínico.

The poisonous snakes of the genera Crotalus and Bothrops have been kept in captivity with the purpose of extracting poison for the production of immunobiological. Knowledge of the physiology of these animals and serum proteins concentration changes are important for early identification of major diseases which lead to states of hypoproteinemia and hyperproteinemia. The objective was to determine the concentration of total protein and serum protein electrophoresis profile of Crotalus durissus terrificus (rattlesnake) in captivity. Blood samples were taken from the ventral coccygeal vein of 21 adult and healthy snakes divided into groups: Group 1 with 12 males, weighing in average 588.89±193.55g, and Group 2 with nine females, weighing in average 708.33±194.04g. The total serum concentration of protein was determined by the method of refractometry and agarose gel electrophoresis. The total protein values in the serum for females was 4.82±0.72, for males 4.51±0.50 and males and females 4.64±0.61, identified by four fractions (g/dL): albumin, a, b and g-globulin. Additionally the albumin/globulin ratio was calculated. The female snakes showed higher values for the variables, albumin and the albumin/globulin (AG) differed significantly (P<0.05) from the group of male snakes, but there was no clinical significance.

Caracterização de Escherichia coli produtoras de toxina de Shiga (STEC) isoladas na produção de bovinos de corte e nas respectivas carcaças dos animais abatidos / Characterization of Shiga toxin-producing Escherichia coli (STEC) isolated in feedlot beef cattle production and their carcasses

Escherichia coli produtoras de toxina de Shiga (STEC) são considerados importantes patógenos de origem alimentar que apresentam o trato intestinal de ruminantes domésticos, principalmente bovinos, seu reservatório natural. Esses microrganismos estão associados com doenças severas em humanos, tais como colite hemorrágica (CH) e síndrome urêmica hemolítica (SHU). Este trabalho teve como objetivos avaliar a ocorrência de STEC em diferentes fontes, ambientais ou não, da criação e abate de bovinos confinados. Além disso, detectar a presença dos genes stx1, stx2, ehxA e eaeA; identificar cepas O157:H7 através da pesquisa do gene uidA; evidenciar a capacidade de produção de Stx e de Eh; identificar variantes de stx e de eaeA; e determinar os sorotipos a diversidade genética das cepas de STEC. A avaliação da presença dos genes (stx1, stx2, ehxA e eaeA) e da produção de Eh foi utilizada como triagem para a seleção de cepas possivelmente patogênicas, sendo que do total de 628 isolados avaliados, foram selecionadas 47 cepas STEC típicas e outras 12 consideradas como atípicas. Das STEC típicas 80,9% foram isolados provenientes de amostras de fezes, enquanto 19,1% foram de amostras de carcaças. Seis cepas isoladas de fezes e 1 de carcaça foram sorotipificadas como 0157:H7, todas positivas para a presença do gene uidA. Além do sorogrupo 0157, nenhum outro, dentre os principais causadores de surtos e casos esporádicos de CH e SHU, foi detectado...

Apo-1 as a marker of programmed cell death in patients with acute lymphoblastic leukaemia

We have planned this work to evaluate the significance and prognostic values of both membrane and soluble APO-1 as markers of apoptosis in patients with acme leukaemia before and alter chemotherapy. For that, 30 patients suffering from acute leukaemia [15 patients with ALL and 15 patients with AMD and 10 apparently healthy individuals serving as control group, were selected and subjected to the following: thorough history and clinical examination, routine investigations including: complete blood picture, bone marrow examination, cytochemistry, immunopheno typing of the blast cells and specific investigations including: detection of mAPO1 [CD95] on surface of blast cells by flow cytometry, detection of DNA fragmentation by agarose gel electrophoresis and measurement of soluble APO-1 by ELISA technique before and after chemotherapy. Surface membrane CD9S was found to be expressed on the majority of ALL blast cells [86.6%] and in only 60% of AML blast cells. The degree of surface membrane expression was variable ranging from 23-86% in ALL and from 43-89%; in AML. In both ALL and AML patients, a significant relationship was detected between surface CD95 expression and response to initial induction chemotherapy. Ninety-one percent of ALL patients and 84% of AML patients who had surface CD95 expression>20% on their blast cells showed complete hematological remission after initial induction chemotherapy. This was confirmed by finding that DNA extracted from patients under chemotherapy, whose blast cells CD95 expression was>20%, showed DNA fragmentation [DNA laddering] by agarose gel electrophoresis [characteristic of apoptosis]. As regards soluble CD9S [SCD95] before starting chemotherapy, no statistically significant difference was observed between the level of soluble CD9S in both ALL and AML patients and the control group [P>0.05]. But, in AML patients, the level of soluble CD95 tended to be etevated [not significantly] in comparison with normal control. After initial induction chemotherapy, the level of soluble CD95 was found to be significantly decreased in both ALL and AML patients in comparison to its level before therapy [P<0.001 and<0.01, respectively]. By following up patients who were resistant to chemotherapy, it was observed that patients who did not achieve complete remission after induction chemotherapy had relatively higher levels of sAPO-1. From these results we can conclude that, since there is a significant relationship between surface CD95 expression in both ALL and AML patients and response to chemotherapy, the expression of surface CD95 could serve as a new prognostic marker as it is helpful in predicting the outcome of therapy. In addition, because soluble APO-1 was found to be relatively high in patients resistant to anti-leukaemic therapy, so measurement of s-APO-1 in sera of acute leukaemia patients could serve as a putative marker for an active persisting leukaemia

Investigation of a possible association between refractory iron deficiency anaemia to an underlying remote helicobacter pylori infection

Helicobacter pylon is a bacterial infection accounts as the prevalent gastric pathogen. Helicobacter has been associated with many extradigestive disorders, as refractory iron deficiency anaemia [Sideropenic]. The aim of this case control study was to investigate the role of remote Helicobacter pylon infection in refractory iron deficiency anaemia [RIDA]; together with comparing two different methods for diagnosis of Helicobacter pylon infection. The study was conducted on thirty patients proved refractory IDA by therapeutic trial. Thirty normal non anaemic subjects were included as controls. Helicobacter pylon testing included stool antigen and Helicobacter pylon PCR. The Helicobacter pylon stool antigen test revealed 12 positive cases out of 30 IDA cases. Five of them were stool PCR cagA positive and four were stool PCR ureC positive. There was 100% agreement between PCR cagA and the stool antigen test in the detection of Helicobacter pylon infection [p=0.003]. Stool PCR cagA had a diagnostic accuracy of 76.67 and likelihood ratio of 3.57. There was 100% agreement between PCR ureC and the stool antigen test in the detection of Helicobacter pylon infection [p=0.009]. Stool FCR ureC had a diagnostic accuracy of 73.33 and likelihood ratio of 3.25. There was a very highly significant difference between the mean 3 of serum ferritin, serum iron, TIBC and transferrin saturation of Helicobacter pylon stool antigen positive and negative subjects [p<0.001]. Conclusion: There was a very highly significant association between Helicobacter pylon infection and refractory iron deficiency anaemia. Serum ferritin levels were significantly lower in Helicobacter pylon stool antigen positive cases than in negative cases [p< 0.001]. Helicobacter pylon positive cases were 2.7 times more likely to develop anaemia than negative cases. The Heticobacterpyioni stool antigen testing by ELISA proved to be more reliable compared to the stool PCR which is tedious and time consuming

Proteinograma sérico de veados-catingueiro (Mazama gouazoubira) criados em cativeiro obtido por eletroforese em gel de agarose e de poliacrilamida (SDS PAGE) / Serum protein concentrations of captive brown brocket deer (Mazama gouazoubira) determined by means of agarosis and sodium dodecyl sulphate-polyacrylamide (SDS-PAGE) gel electrophoresis

The serum protein concentrations of brown brocket deer (Mazama gouazoubira) obtained by agarosis gel and sodium dodecyl sulphate-polyacrylamide (SDS-PAGE) gel were determined from blood samples of ten adult healthy animals (six females and four males), monthly collected in the morning, during 12 months. The animals, maintained in individual stable and protected from noise, received ad libitum a diet of comercial ration and green roughage. Serum protein concentrations in agarosis gel revealed the presence of four protein fractions: albumin, alphaglobulin, betaglobulin, and gammaglobulin. Only serum concentrations of albumin were influenced by season, being values in spring higher than values in summer (4.15 x 3.64g/dl). Serum concentrations of albumin (4.05 x 3.75g/dl) were higher for female and alphaglobulin (0.39 x 0.53g/dl) werehigher for males. Results showed 34 proteins with molecular weights ranging from 18kD to 165kD. Significant differences between at least two seasons were found on values of 11 proteins. In conclusion, on account of the 10 animals been maintained in the same physical space and submitted to the same handling system, physiological variations, which are characteristic of this species, can be apointed as the reason of these differences.

Increased isolation and characterization of Shigella sonnei obtained from hospitalized children in Tehran, Iran.

Shigella flexneri has been the most frequent cause of shigellosis in children in Iran. To evaluate the changes in frequency of serogroups, 302 Shigella species were isolated in 2003 from hospitalized children, aged less than 12 years, with acute diarrhoea in Tehran, Iran. The number of collected S. sonnei, S. flexneri, S. boydii, and S. dysenteriae isolates was 178 (58.9%), 110 (37.4%), 10 (3.3%), and 4 (1.3%) respectively. Most (94%) S. sonnei isolates were resistant to co-trimoxazole. They were, however, relatively or completely sensitive to 15 commonly-used antibiotics. The extracted plasmids showed 12 different profiles with two closely-related patterns constituting 70% of the total isolates. Ribotyping, using PvuII, HindIII or SalI restriction enzymes, generated a single pattern for all S. sonnei isolates. Data suggest that S. sonnei has become the predominant serogroup in children in the hospitals of Tehran.

Screening of RET gene mutations in multiple endocrine neoplasia type-2 using Conformation Sensitive Gel Electrophoresis (CSGE)

Multiple endocrine neoplasia type 2 (MEN2) is an autosomal dominant inherited tumor syndrome caused by RET proto-oncogene germline mutations (RET). Here we tested the Conformation Sensitive Gel Electrophoresis (CSGE) as a screening method for RET hot-spot mutations. Seven MEN2 families were studied by direct sequencing analysis, CSGE and Single Strand Conformational Polymorphism (SSCP). Using CSGE/SSCP, we were able to detect four out of five types of RET mutations verified by sequencing analysis: Cys620Arg, Cys634Arg, Cys634Tyr, and Met918Thr, furthermore a missense substitution at codon 648 (Val648Ile). RET polymorphisms 691 and 769 were also verified. Data obtained using CSGE/SSCP were fully concordant. We conclude that CSGE showed to be a sensitive, fast, low-cost, and simple procedure to detect RET mutations in codons which are reported as the most prevalent RET variants (~ 95 percent) in large MEN2 series. As to the Val804Met mutation, this method still needs to be optimized.

A neoplasia endócrina múltipla tipo 2 (NEM2) é uma síndrome tumoral herdada por mutações germinativas no proto-oncogene RET (RET). Analisamos a aplicação do método Eletroforese em Gel Sensível à Conformação (CSGE) no rastreamento de mutações hot spots do RET. Sete famílias com NEM2 foram rastreadas pelo seqüenciamento gênico, CSGE e análise do Polimorfismo Conformacional de Cadeia Simples (SSCP). Usando ambas as metodologias de rastreamento, identificamos quatro dos cinco tipos de mutações verificadas pelo seqüenciamento: Cys620Arg, Cys634Arg, Cys634Tyr e Met918Thr, além da variação gênica Val648Ile. Das análises englobando mutações hot spots do RET, 90,6 por cento concordaram com o seqüenciamento genético (incluindo a variação gênica Val648Ile). Polimorfismos nos códons 691 e 769 foram documentados. Os dados obtidos por CSGE/SSCP foram totalmente concordantes. Concluímos que o CSGE revelou ser metodologia sensível, rápida, de fácil execução e baixo custo no rastreamento de mutações nos códons associados à grande maioria (~ 95 por cento) dos pacientes com NEM2.

Detection of beta-thalassemia (homozygous) by hemoglobin electrophoresis on agar gel and citrate agar medium: a case report.

A seven month old male baby was admitted to NICU of BPKIHS, Dharan with dyspnoea and distress. He was investigated for hemolytic anemia and suspected of beta Thalassemia (Homozygous) based on the low level of Hb and high HbF%. To investigate further, Hb electrophoresis was carried out using agar gel and citrate agar gel at pH 8.6 and 6.0 respectively along with control. Electrophoretogram showed single narrow band lagging behind the healthy control moved toward anode at pH 8.6 and single narrow band moved parallel to healthy control toward cathode at pH 6.0. Thus the diagnosis of betaThalassemia (Homozygous) is favored.

Apoptosis of cultured cortical neurons of rat's brain induced by heroin / 法医学杂志

OBJECTIVE@#To investigate whether heroin can directly induce apoptosis in primary cultured cortical neurons of rat's brain.@*METHODS@#Cultured primary neurons cultures were obtained from cerebral cortex of embryo rats. After 7 days, the cells were incubated with different concentrations of heroin (purity-80%) for 24 hours. The neuronal survival was assessed by cell viability counting with fluorescent diacetate (FDA) staining. The morphological and biochemical changes were observed with Hoechst 33258 fluorescent staining and then analyzed by agarose gel electrophoresis, respectively.@*RESULTS@#After treatment with different concentrations of heroin, the neurons showed a decreased survival rate in a dose dependent manner, and there was a significant difference in the survival rate between the heroin group and the control group (P < 0.05). When exposed to different concentrations of heroin, neurons exhibited the morphological and biochemical features of apoptosis, including cell shrinkage, neurite degeneration, network disappearance, condensation and aggregation of nuclear chromatin, and the formation of DNA ladders. With the increase of heroin concentration of rat's brain more apoptotic bodies were seen.@*CONCLUSION@#Heroin can directly induce apoptosis in primary cultured cortical neurons in rat's brain.