PLACENTAL ALKALINE PHOSPHATASE AS A THRESHOLD CONCEPT FOR EARLY DETECTION OF PRIMARY LUNG CANCER

The biochemical existing tool of diagnostic methods to lung cancer cases need to be improved. In order to validate an early screening of primary tumor patients, a developed a simple procedure or technique was demanded. The aims of this study were to provide an overview of alkaline Placental Alkaline Phosphatase activity in lung cancer. Using heating inactivation method regarding the measurement of Placental Alkaline Phosphatase activity as an early diagnosis marker in lung cancer cases. Total alkaline phosphatase and Placental alkaline phosphatase activity were measured in patients of Lung cancer patients who were classified according to the site of tumor by histological picture. ALP isoenzymes were identified by heat inactivation, and compared with the most frequently applied method (ELISA). Monitoring of the Total ALP and Placental ALP activity in the studied groups using two different methods were shown a highly performance of heating method by an experimental assessment to confirm the accuracy and validity of the proposed method. The distribution of serum placental ALP isoenzyme activity in patients and control groups which was measured by two different methods were found to be (20.2-43.1) IU/L respectively (measured by heating method) and (394.3- 454.5) pg/mL measured by ELISA method) respectively. Placental ALP isoenzyme showed a high significant activity in lung cancer patients than healthy control with p value less than (0.05). That application of the heat inactivation method yields similar indication to the ones obtained by the highly and specific enzyme-linked immunosorbent assay. The results of detection Placental alkaline phosphatase in serum were in excellent agreement and could have a potentially extensive application for Placental alkaline phosphatase quantification.

Electroforesis de isoenzimas de fosfatasa alcalina: utilidad diagnóstica en pacientes con enfermedad de células falciformes / Alkaline phosphatase isoenzyme electrophoresis: diagnostic utility in patients with sickle cell disease

Introducción: Las fosfatasas alcalinas (FAL) humanas son indicadores enzimáticos de daño de órganos. Las complicaciones de la drepanocitosis incluyen lesión de órganos debido a la reperfusión, la vasculopatía proliferativa y la anemia hemolítica. Objetivo: Demostrar la utilidad de la determinación de la actividad enzimática de la FAL y la movilidad electroforética de las isoenzimas séricas de FAL en pacientes con drepanocitosis en estado basal y durante las crisis. Métodos: Estudio descriptivo, longitudinal, prospectivo en 85 individuos adultos, 25 controles sanos y 60 pacientes, entre enero y diciembre/2018. Variables estudiadas: genotipos (SS/Sβo, SC/Sβ+), edad, sexo, actividad de FAL e isoenzimas por electroforesis en gel de agarosa con y sin lectina: hepática 1 (H1) y (H2); placenta 1(P1), ósea, intestinales 1, 2, 3 (I1.2.3) en estado basal y crisis. Resultados: La actividad de la FAL fue significativamente mayor en los pacientes que en los controles. Hubo diferencias significativas en la actividad de la FAL y de la fracción H1+P1/1 de pacientes en estado basal en relación con el grupo control. La actividad de la enzima y las isoenzimas no mostró diferencias significativas entre los genotipos. Igual comportamiento se observó en la actividad de las enzimas e isoenzimas durante las crisis vasoclusivas dolorosas y hepáticas. Se encontraron diferencias significativas en la actividad de la fracción hepática H1+P1/1 entre los grupos de 20-29 y 40-49 años Conclusiones: La determinación de la FAL en los pacientes con drepanocitosis es útil y permite establecer un perfil isoenzimático personalizado, con importancia pronóstica como un marcador biológico de alarma(AU)

Introduction: Human alkaline phosphatases (ALP) are enzymatic indicators of organ damage. Complications of sickle cell disease include injury to organs due to reperfusion injury, proliferative vascular disease, and hemolytic anemia. Objective: To demonstrate the usefulness of determing ALP activity and the electrophoretic mobility of the serum ALP isoenzymes in patients with sickle cell disease at base line and during the crises. Methods: A descriptive, longitudinal, prospective study was carried out in 85 adult individuals, 25 healthy controls and 60 patients, between January and December/2018. Studied variables: genotypes (SS/SβO, SC/Sβ+), age, sex, ALP activity and isoenzymes by agarose gel electrophoresis with and without lectin: hepatic 1 (H1) and (H2); placenta 1 (P1), bone, intestinal 1, 2, 3 (I1.2.3) in basal state and crisis. Results: ALP activity was significantly higher in patients than in controls. Significant differences were found in the activity of the ALP and fraction H1+P1/1 of patients in basal state in relation to the control group. The activity of the enzyme and the isoenzymes showed no significant differences between genotypes. The same behavior was observed in the activity of enzymes and isoenzymes during painful and liver vasooclusive crises. Significant differences were found in the activity of the liver fraction H1+P1/1 between the groups of 20-29 and 40-49 years. Conclusions: The determination of the ALP in patients with sickle cell disease is useful and allows to establish a personalized isoenzimatic profile, with prognostic importance as a biological alarm marker(AU)

Testicular germ cell tumors type 2 have high RNA expression of / 亚洲男科学杂志(英文版)

This study analyzed RNA expression of genes for three serum tumor markers, alpha fetoprotein (AFP), human chorionic gonadotropin (hCG), and lactate dehydrogenase (LDH), in patients with testicular germ cell tumors (TGCT) type 2. The gene AFP encodes AFP, the gene for chorionic gonadotropin beta polypeptide 5 (CGB5) encodes a major part of the specific beta subunit of hCG, and the genes for LDH subunit A (LDHA), LDH subunit B (LDHB), and LDH subunit C (LDHC) encode three different subunits of LDH. LDHB encodes the LDHB subunit present as a tetramer in LDH isoenzyme 1 (LDH-1). We examined three datasets with 203 samples of normal testis tissue (NT) and TGCT type 2. Yolk sac tumor (YST) expressed RNA of AFP fourteen thousand times higher than seminoma (SE), embryonal carcinoma (EC), and teratoma (TER) combined (P = 0.00015). In the second microarray, choriocarcinoma (CC) expressed RNA of CGB5 ten times higher than other histologic types of TGCT combined. EC expressed RNA of LDHB twice higher than SE, YST and TER combined (P = 0.000041). EC expressed RNA of LDHB higher than that YST expressed RNA of AFP and that CC expressed RNA of CGB5. In conclusion, TGCT type 2 expressed RNA of LDHB markedly higher than the RNA of 23 other candidate genes for TGCT type 2.

Germination temperatures affect the physiological quality of seeds of lettuce cultivars / Qualidade fisiológica de sementes de alface sob diferentes temperaturas na germinação

The objective of this study was to evaluate the physiological quality allied to biochemical quality of lettuce seeds by germination and enzymes expression at 20, 25, 30, 35, 40 and 42ºC. Germination speed index and percentage of germination were estimated. Isoenzyme expressions were assessed by alcohol dehydrogenase (ADH), malate dehydrogenase (MDH), catalase (CAT), esterase (EST), pyruvate decarboxylase (PDC) and glutamate oxaloacetate transferase (GOT). The experiment consisted of a completely randomized design in a factorial scheme 4x6, with four cultivars and six different temperatures, with four replications. The highest germination and vigor were observed for cv. 'Everglades' at 35°C, which proved that this cultivar is thermotolerant. Catalase can be considered a genetic marker for the identification ofthermotolerant lettuce cultivars. Cultivar 'Everglades' has potential to be used in lettuce breeding programs aimed at cultivars tolerant to high temperatures during germination.

O objetivo deste estudo foi avaliar a qualidade fisiológica e bioquímica de sementes de alface por meio da germinação e expressão de enzimas a 20, 25, 30, 35, 40 e 42ºC. As variáveis velocidade de germinação e o índice de velocidade de germinação foram estimadas. As expressões das enzimas alcool desidrogenase (ADH), malato desidrogenase (MDH), catalase (CAT), esterase (EST), piruvate descarboxilase (PDC) e glutamato oxaloacetato transferase (GOT) foram avaliadas. Para análise dos genótipos foi empregado o delineamento inteiramente casualizado em esquema fatorial 4x6, testando quatro cultivares e seis diferentes temperaturas, com quatro repetições. A maior germinação e vigor foram observadas para a cv. 'Everglades' a 35°C, o que prova que esta cultivar é termotolerante. A catalase pode ser considerada um marcador para a identificação de cultivares de alface termotolerantes. A cultivar 'Everglades' tem potential para uso em programas de melhoramento visando tolerância à alta temperatura durante a germinação.

Effect analysis of diazepam combined with phenobarbital in the treatment of febrile convulsions in children / 中国基层医药

Objective@#To investigate the clinical effect of diazepam combined with phenobarbital in the treatment of febrile convulsions in children.@*Methods@#From January 2015 to July 2017, 100 children with febrile convulsions in the Women's and Children's Hospital of Ningbo were selected and divided into two groups by random number table, with 50 cases in each group.The control group was treated with diazepam, while the observation group was treated with diazepam combined with phenobarbital.The clinical efficacy, anticonvulsive time, myocardial enzyme index, hypersensitive C-reactive protein and incidence of adverse reactions were compared between the two groups.@*Results@#(1)The total clinical effective rate of the observation group(98%) was higher than that of the control group(χ2=4.891, P<0.05), and the time of stopping convulsion[(20.92±4.49)min] was shorter than that of the control group(t=5.443, P<0.05). (2)After treatment, the creatine kinase isoenzymes[(25.93±12.76)U/L], phosphocreatinase[(189.47±7.35)U/L], hydroxybutyrate dehydrogenase[(276.73±12.85)U/L], lactate dehydrogenase[(305.94±13.27)U/L], hypersensitive C-reactive protein[(5.04±1.65)ng/L] in the observation group were lower than those in the control group(t=4.961, 5.333, 5.901, 6.160, 5.354, all P<0.05). (3)During the treatment period, no obvious adverse reactions occurred in both two groups.@*Conclusion@#The combination of diazepam and phenobarbital can effectively arrest febrile convulsions in children, improve the clinical efficacy, protect myocardial cells, it has fewer adverse reactions, it is safe and reliable.

Effect analysis of diazepam combined with phenobarbital in the treatment of febrile convulsions in children / 中国基层医药

To investigate the clinical effect of diazepam combined with phenobarbital in the treatment of febrile convulsions in children.Methods From January 2015 to July 2017,100 children with febrile convulsions in the Women＇s and Children＇s Hospital of Ningbo were selected and divided into two groups by random number table,with 50 cases in each group.The control group was treated with diazepam ,while the observation group was treated with diazepam combined with phenobarbital.The clinical efficacy,anticonvulsive time,myocardial enzyme index,hypersensitive C-reactive protein and incidence of adverse reactions were compared between the two groups . Results (1)The total clinical effective rate of the observation group (98%) was higher than that of the control group (χ2 ＝4.891,P＜0.05),and the time of stopping convulsion [(20.92 ±4.49) min] was shorter than that of the control group(t＝5.443,P＜0.05).(2) After treatment,the creatine kinase isoenzymes [(25.93 ±12.76) U／L], phosphocreatinase [(189.47 ±7.35 ) U／L], hydroxybutyrate dehydrogenase [( 276.73 ±12.85 ) U／L ], lactate dehydrogenase[(305.94 ±13.27)U／L],hypersensitive C-reactive protein[(5.04 ±1.65)ng／L] in the observation group were lower than those in the control group (t＝4.961,5.333,5.901,6.160,5.354,all P＜0.05).(3) During the treatment period, no obvious adverse reactions occurred in both two groups.Conclusion The combination of diazepam and phenobarbital can effectively arrest febrile convulsions in children ,improve the clinical efficacy ,protect myocardial cells,it has fewer adverse reactions ,it is safe and reliable.

Diagnostic value of lactate dehydrogenase isoenzymes in children with refractory mycoplasma pneumoniae pneumonia / 中国医师进修杂志

Objective To investigate the diagnostic value of lactate dehydrogenase (LDH) isoenzymes in children with refractory mycoplasma pneumoniae pneumonia (RMPP). Methods One hundred and forty-seven children with mycoplasma pneumoniae pneumonia (MPP) in Children′s Medical Center, Dongfeng Hospital Affiliated to Hubei University of Medicine from July 2016 to July 2018 were selected. Among them, RMPP was in 54 cases (RMPP group), and non-RMPP was in 93 cases (non-RMPP group). The white blood cell count (WBC), C- reactive protein (CRP), erythrocyte sedimentation rate (ESR), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatine kinase (CK) were detected; meanwhile, the serum levels of total LDH and LDH isoenzymes (LDH1, LDH2, LDH3, LDH4 and LDH5) were measured by radioimmunoassay. Results The ESR, AST, ALT, LDH1, LDH2, LDH4, LDH5 and total LDH in RMPP group were significantly higher than those in non-RMPP group:(23.15 ± 3.62) mm/h vs. (19.45 ± 3.04) mm/h, (37.20 ± 6.82) U/L vs. (31.49 ± 4.70) U/L, (35.48 ± 5.72) U/L vs. (27.31 ± 7.22) U/L, (89.77 ± 5.26) U/L vs. (85.01 ± 7.65) U/L, (154.60 ± 22.30) U/L vs. (133.17 ± 32.82) U/L, (61.51 ± 10.09) U/L vs. (47.74 ± 11.22) U/L, (78.99 ± 12.86) U/L vs. (64.98 ± 7.14) U/L and (511.49 ± 38.90) U/L vs. (450.19 ± 47.63) U/L, and there were statistical differences (P＜0.05); there were no statistical differences in WBC, CRP, CK and LDH3 between 2 groups (P＞0.05). Logistic regression analysis result showed that the ESR, AST, ALT, LDH1, LDH4 and LDH5 were the risk factors of RMPP ( OR=0.618, 0.735, 0.785, 0.769, 0.866, 0.880; 95% CI 0.430 to 0.888, 0.596 to 0.906, 0.665 to 0.926, 0.646 to 0.917, 0.781 to 0.961, 0.803 to 0.965; P<0.01). Receiver operating characteristic curve analysis result showed that the area under curve (AUC) of RMPP predicted by LDH5 was the largest (0.857), and then by LDH4 (0.819). The critical values were 70.74 and 53.14 U/L respectively; the AUC of RMPP predicted by LDH4 combined with LDH5 was 0.914, and the critical values was 130.10 U/L, with a sensitivity of 83.3% and a specificity of 92.5% , 95% CI 0.856 to 0.954. Conclusions Serum levels of LDH4 and LDH5 are significantly increased in children with RMPP. LDH4 combine with LDH5 detection might be useful in predicting the occurrence of RMPP.

Molecular typing of Leishmania (Leishmania) amazonensis and species of the subgenus Viannia associated with cutaneous and mucosal leishmaniasis in Colombia: A concordance study / Tipificación molecular de Leishmania (Leishmania) amazonensis y especies del subgénero Viannia asociadas a la leishmaniasis cutánea y la mucosa en Colombia: un estudio de concordancia

Resumen Introduction: Multilocus enzyme electrophoresis (MLEE) is the reference standard for the characterization of Leishmania species. The test is restricted to specialized laboratories due to its technical complexity, cost, and time required to obtain results. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is used to identify Leishmania species. Objective: To establish the concordance between the two tests as identifying methods for circulating species in Colombia. Materials and methods: A total of 96 isolates from patients with cutaneous or mucosal leishmaniasis were selected and identified by MLEE and PCR-RFLP with miniexon and hsp70 as the molecular targets, which were used sequentially. Restriction enzymes HaeIII and BccI were similarly applied. Cohen's kappa coefficient and the 95% confidence interval (CI) were calculated. Results: The kappa coefficient and the 95% CI between MLEE and PCR-RFLP displayed "very good" concordance with a coefficient of 0.98 (CI95%: 0.98 to 1.00). The identified species were Leishmania Viannia braziliensis, Leishmania Viannia panamensis, Leishmania Viannia guyanensis and Leishmania Leishmania amazonensis. A total of 80 of the 96 isolates were sequenced and the results obtained by PCR-RFLP were confirmed. Conclusion: Due to the concordance obtained between tests results with the amplification of the genes miniexon and hsp70, PCR-RFLP is proposed as an alternative for identifying circulating Leishmania species in Colombia.

Abstract Introducción. La electroforesis de enzimas multilocus (Multilocus Enzyme Electrophoresis, MLEE) es el estándar de referencia para la tipificación de las especies de Leishmania. La prueba está restringida a laboratorios especializados por su complejidad técnica, sus costos y el tiempo necesario para obtener resultados. La PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) se utiliza para tipificar especies de Leishmania. Objetivo. Establecer la concordancia entre las dos pruebas como métodos de tipificación de las especies circulantes en Colombia. Materiales y métodos. Se seleccionaron 96 aislamientos de pacientes con leishmaniasis cutánea o mucocutánea y se tipificaron mediante MLEE y PCR-RFLP con los blancos moleculares miniexon y hsp70 usados en serie. Las enzimas de restricción aplicadas fueron la HaeIII y la BccI, respectivamente. Se calculó el coeficiente kappa y un intervalo de confianza (IC) de 95 %. Resultados. Se determinó que la concordancia fue "muy buena" al obtener un coeficiente de 0,98 (IC95%: 0,98-1,00). Las especies identificadas fueron: Leishmania Viannia braziliensis, L. (V.) panamensis, L. (V.) guyanensis y L. (L,) amazonensis. De los 96 aislamientos, 80 se enviaron a secuenciación y se confirmaron los resultados obtenidos mediante PCR-RFLP. Conclusión. Dada la concordancia obtenida con la PCR-RFLP amplificando los genes miniexon y hsp70, se propone esta prueba como alternativa para la tipificación de especies de Leishmania circulantes en Colombia.

Total Creatine Kinase And Its Isoenzymes In Cord Blood Of Full Term Newborns

Creatine kinase (CK) is a key enzyme of energy metabolism, especially in muscle tissue. CK has two polypeptide chains of M and B, and three isomers, CK-BB, CK-MB and CK- MM. In some conditions like acute myocardial infarction and Neuromuscular disorders, increased CK activity is used as a part of diagnosis. CK can also be elevated in absence of neuromuscular diseases or cardiac injury, such as strenuous exercise, intramuscular injections etc.Several reports indicate elevated activity of serum CK in the maternal blood during child birth.

Hiperfosfatasemia transitoria benigna de la infancia: Serie clínica / Benign transient hyperphosphatasaemia in infants: Clinical series

Las fosfatasas alcalinas (FA) son un grupo de 4 isoenzimas que se producen en diversos tejidos, pudiendo elevarse en condiciones fisiológicas y secundariamente a enfermedades óseas o hepatobiliares. En niños una de las causas más frecuentes e inocuas, pero poco conocidas, es la hiperfosfatasemia transitoria benigna de la infancia (HFTBI). El objetivo es reportar una serie de casos de HFTBI y proponer un criterio de enfrentamiento. Casos clínicos: Se presentan 5 niños de entre 11 y 50 meses de edad, 4 de ellos de sexo femenino, con hallazgo incidental de elevación severa (> 1.000 UI/l) en los niveles de FA en exámenes solicitados por mal incremento pondo-estatural o por cuadros infecciosos. A través de la anamnesis, examen físico y laboratorio básico se descartó enfermedad ósea o hepática. En 2 de los pacientes se determinaron las isoenzimas, destacando el predominio óseo. Se comprobó una normalización de los niveles de FA en un periodo de uno a 6 meses, sin evidencia de complicaciones posteriores. Conclusión: La HFTBI es un desorden bioquímico benigno de evolución autolimitada, que es importante tener presente al enfrentar un niño menor de 5 años con elevación severa de FA, en ausencia de alteraciones clínicas o de laboratorio que sugieran enfermedad ósea o hepática.

Alkaline Phosphatase (ALP) is a group of 4 isoenzymes produced in different tissues. Elevated levels of ALP can be developed under physiological conditions, and can indicate the presence of bone or hepatobiliary diseases. In children, one of its most common harmless causes is benign transient hyperphosphatasaemia (BTH), a little known condition. The objective is to report BTH cases and propose a monitoring plan. Case reports: The cases of 5 children aged between 11 and 50 months are presented, 4 of them female, with the incidental finding of a sudden and severe ALP elevation (> 1,000 U/l), in tests ordered due to either abnormal growth and development, or intercurrent infections. Bone and liver disease were ruled out using the patient history, physical examination and basic laboratory results. Isoenzymes levels were determined in 2 patients. A return to normal ALP levels was observed over a period of 1-6 months, with no evidence of further complications. Conclusion: BTH is a benign self-limiting biochemical disorder, which should be considered in children under 5 years old with severe ALP elevation in the absence of clinical or laboratory abnormalities suggestive of bone or liver disease.

Effects of lysine and gluconic acid zinc granules combined with cimetidine on serum creatine kinase isoenzyme, aspartate aminotransferase and interleukin-6 levels and prognosis in children with rotavirus enteritis / 中国医师进修杂志

Objective To investigate the effects of lysine and gluconic acid zinc granules combined with cimetidine on serum creatine kinase isoenzyme (CK-MB), aspartate aminotransferase (AST) and interleukin-6 (IL-6) levels and prognosis in children with rotavirus enteritis. Methods Eighty-one children with rotavirus enteritis were randomly divided into control group (40 patients) and observation group (41 patients).The patients in control group were treated with cimetidine, and those in the observation group were treated with cimetidine combined with lysine and gluconic acid zinc granules. Clinical treatment, dehydration correction time, diarrhea time, fever time and serum CK-MB, AST and IL-6 levels were observed and compared between two groups. The occurrence of toxic side effects and vitality, physiological function, mental status scores were statistically analyzed. Results The total effective rate in observation group was 95.12%(39/41), in control group was 75.00%(30/40), and there was significant difference (P < 0.05). The dehydration correction time, diarrhea time, fever time after treatment in observation group was less than that in control group (P<0.05). The serum CK-MB, AST and IL-6 levels after treatment in observation group were significantly lower than those in control group (P<0.01). There was no significant difference in the incidence of side effects between observation group 17.07%(7/41) and control group 10.00%(4/40) (P>0.05). The scores of physiological function, vitality and mental status scores in observation group were higher those in control group, and there were significant differences(P < 0.01). Conclusions The lysine and gluconic acid zinc granules combined with cimetidine in the treatment of rotavirus enteritis gets has a significant effect and high safety, which can alleviate the inflammatory reaction and improve the myocardial enzymes levels and the quality of life.

Effects of lysine and gluconic acid zinc granules combined with cimetidine on serum creatine kinase isoenzyme, aspartate aminotransferase and interleukin-6 levels and prognosis in children with rotavirus enteritis / 中国医师进修杂志

Objective To investigate the effects of lysine and gluconic acid zinc granules combined with cimetidine on serum creatine kinase isoenzyme (CK-MB), aspartate aminotransferase (AST) and interleukin-6 (IL-6) levels and prognosis in children with rotavirus enteritis. Methods Eighty-one children with rotavirus enteritis were randomly divided into control group (40 patients) and observation group (41 patients).The patients in control group were treated with cimetidine, and those in the observation group were treated with cimetidine combined with lysine and gluconic acid zinc granules. Clinical treatment, dehydration correction time, diarrhea time, fever time and serum CK-MB, AST and IL-6 levels were observed and compared between two groups. The occurrence of toxic side effects and vitality, physiological function, mental status scores were statistically analyzed. Results The total effective rate in observation group was 95.12%(39/41), in control group was 75.00%(30/40), and there was significant difference (P < 0.05). The dehydration correction time, diarrhea time, fever time after treatment in observation group was less than that in control group (P<0.05). The serum CK-MB, AST and IL-6 levels after treatment in observation group were significantly lower than those in control group (P<0.01). There was no significant difference in the incidence of side effects between observation group 17.07%(7/41) and control group 10.00%(4/40) (P>0.05). The scores of physiological function, vitality and mental status scores in observation group were higher those in control group, and there were significant differences(P < 0.01). Conclusions The lysine and gluconic acid zinc granules combined with cimetidine in the treatment of rotavirus enteritis gets has a significant effect and high safety, which can alleviate the inflammatory reaction and improve the myocardial enzymes levels and the quality of life.

Purification and characterization of buffalo liver L-arginase and its kinetic properties with dihydropyrimidine and metal ions.

Arginase (L-arginine amidinohydrolase, EC.3.5.3.1) from animal tissues such as, liver and kidney has been partially characterized by many researchers. In this study, we purified arginase to homogeneity from buffalo liver with about ~2857 purification fold and a 20% recovery by chromatographic and spectroscopic analysis were obtained. The molecular mass determined by gel filtration and SDS-PAGE was found to be 118 kDa and 47 kDa, respectively. The optimal pH and temperature of the arginase was 9.5 and 40°C, respectively. Kinetic parameters (Km and Vmax) showed activation of arginase in the reaction medium with decrease in Km (7.14, 5.26, 4.0 and control 3.22 mM) and Vmax (0.05, 0.035, 0.027 and control 0.021 mg/mL/min), while co-factor activity of arginase was optimized using metal ions like Mn2+ and Mg2+ at 2 mM, which revealed an increase in Vmax values (0.011, 0.013, 0.015 and control 0.010 mg/mL/min) and a decrease in Km values (2.22, 2.12, 1.88 and control 1.66 mM). The kinetic data suggested that the arginase activity is enhanced in the presence of dihydropyrimidine derivative and metal ions, indicating essential mode of activation.

Significance of serum total alkaline phosphatase levels in breast cancer

Background: Breast cancer has shown an increased prevalence worldwide, both in industrialized and developing countries. Unfortunately, most cancers do not produce any symptoms until the tumors are either too large to be removed surgically or metastasis has taken place. Therefore, there is need for simple biochemical investigations, which can be easily assayed, less expensive and can predict stages and prognosis. This study was undertaken to know the usefulness of estimation of serum Alkaline Phosphatase (ALP) in assessing the prognosis and staging of breast cancer in smaller laboratories where sophisticated technology is not available. Materials & Methods: A total number of 95 subjects participated in the present study which included 60 breast cancer cases and 35 controls. Serum Alkaline Phosphatase levels were estimated in preoperative & postoperative breast cancer subjects of different stages and control. ALP Levels were compared between control and breast cancer. Also comparison was done between different stages and further, ALP levels compared pre and post operatively. Results: Rise in the ALP was found in breast cancer when compared to controls. ALP levels increased significantly with the stage of cancer. Postoperatively, there was significant decrease in ALP levels in each stage. Conclusion: Though less sensitive than imaging procedures, measurement of serum ALP is cost effective and may be useful in smaller laboratories for staging, to know the prognosis and monitor the treatment. Serial analysis using plasma Alkaline phosphatase isoenzymes combined with other parameters like Gamma glutamyl transferase (GGT) for the detection of metastasis would seem to be justified.

Clinical and preclinical treatment of urologic diseases with phosphodiesterase isoenzymes 5 inhibitors: An update / 亚洲男科学杂志(英文版)

Phosphodiesterase isoenzymes 5 inhibitors (PDE5-Is) are the first-line therapy for erectile dysfunction (ED). The constant discoveries of nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) cell-signaling pathway for smooth muscle (SM) control in other urogenital tracts (UGTs) make PDE5-Is promising pharmacologic agents against other benign urological diseases. This article reviews the literature and contains some previously unpublished data about characterizations and activities of PDE5 and its inhibitors in treating urological disorders. Scientific discoveries have improved our understanding of cell-signaling pathway in NO/cGMP-mediated SM relaxation in UGTs. Moreover, the clinical applications of PDE5-Is have been widely recognized. On-demand PDE5-Is are efficacious for most cases of ED, while daily-dosing and combination with testosterone are recommended for refractory cases. Soluble guanylate cyclase (sGC) stimulators also have promising role in the management of severe ED conditions. PDE5-Is are also the first rehabilitation strategy for postoperation or postradiotherapy ED for prostate cancer patients. PDE5-Is, especially combined with adrenoceptor antagonists, are very effective for benign prostatic hyperplasia (BPH) except on maximum urinary flow rate (Q max ) with tadalafil recently proved for BPH with/without ED. Furthermore, PDE5-Is are currently under various phases of clinical or preclinical researches with promising potential for other urinary and genital illnesses, such as priapism, premature ejaculation, urinary tract calculi, overactive bladder, Peyronie′s disease, and female sexual dysfunction. Inhibition of PDE5 is expected to be an effective strategy in treating benign urological diseases. However, further clinical studies and basic researches investigating mechanisms of PDE5-Is in disorders of UGTs are required.

On the origin and diversification of Venezuelan freshwater fishes: the genus Gephyrocharax (Ostariophysi: Characidae) a case study

We conducted a phylogeographic analysis of the genus Gephyrocharax in Venezuela to evaluate geomorphologic evidence for the formation of the country's main watersheds and to establish a biogeographical hypothesis of possible diversification mechanisms of the Neotropical freshwater fish fauna. We assayed eight enzyme systems and general proteins to estimate genetic variability (H, P), intraspecific structuring in several Gephyrocharax valencia and G. venezuelae populations (FIS, FIT, and FST), and a phylogenetic approach for the three species of Gephyrocharax in Venezuela, using Corynopoma riisei as the external group. Fourteen presumptive loci indicate that populations of the three species of Gephyrocharax analyzed show a clear genetic inter-specific differentiation, determined by four loci with fixed alleles (GPI-B*, IDH*, ME-1*, and ME-2*). The resulting cladogram shows two major clades: a monophyletic group consisting of Gephyrocharax n. sp. and G. venezuelae (restricted to the northwest of the country) and a group formed exclusively by G. valencia (distributed along the largest geographic range). Speciation of the Venezuelan lineages of the genus Gephyrocharax could be explained by the origin and course movements of the present Orinoco River together with geomorphologic processes that have occurred in northern Venezuela since the Miocene.

Foi feita uma análise filogeográfica do gênero Gephyrocharax na Venezuela a fim de avaliar as evidências geomorfológicas que levaram à formação dos principais sistemas hidrográficos do país, além de estabelecer uma hipótese biogeográfica com os possíveis mecanismos de diversificação da fauna de peixes de água doce Neotropical. Foram analisados oito sistemas enzimáticos e proteínas gerais para conhecer a variabilidade genética (H, P), estruturação intraespecífica em populações de Gephyrocharax valencia e G. venezuelae (FIS, FIT e FST), e uma abordagem filogenética com base na análise isozimática para as três espécies de Gephyrocharax na Venezuela, com Corynopoma riisei como grupo externo. Quatorze loci presumíveis indicam que as populações das três espécies de Gephyrocharax analisados revelam uma diferenciação inter-específica genética, determinada por quatro loci com alelos fixos (GPI-B*, IDH*, ME-1* e ME-2*). O cladograma resultante apresenta dois clados principais: um grupo monofilético composto por Gephyrocharax n. sp. e G. venezuelae (restrita ao noroeste do país) e um grupo formado exclusivamente por G. valencia (distribuídos ao longo da maior área geográfica). A especiação das linhagens de Gephyrocharax na Venezuela poderia ser explicada pela origem e movimentos do curso atual da bacia do rio Orinoco, associado a processos geomorfológicos que ocorrem no norte da Venezuela desde o Mioceno.

Genetic Analysis of Izoenzymes Polymorphisms in Silkworm (Bombyx mori L) Strains / Análise genética de polimorfismos bioquímicos em linhagens do bicho da seda (Bombyx mori L)

This work carried out to evaluates the polymorhism in the silkworm of different lineages using the isoenzymes electrophoresis to detect biochemical markers and to investigate the genetics of populations for those lineages. They were used as samples individual extracts of silk glands of second day old larvas of the fifth instar, originating from seven Japanese lineages and eight pure Chinese lineages maintained by the Cocamar- Cooperativa Agroindustrial de Maringá. The isozymes acid phosphatase (ACP), alkaline phosphatase (AKP) and carbonic anhydrase (CA) they were submitted to the electrophoresis in starch gels 14%. The esterases (EST) were analyzed in polyacrylamide gels to 10% and stained with α and ß-naphtyl acetate. The total of 21 loci was detected, and 04 (19.05%) they are polymorphic, Est-11, Acp, Akp, Ca. The fixation index (Fis) for the analyzed isozymes it was 0.0751, indicating excess of homozygotes. The value of Fst (0.6128) it shows that the lineages are well differentiated. The dendrogram obtained with the values of genetic distance didn't separate the Chinese and Japanese lineages analyzed totally. That preliminary evaluation of the lineages of B. mori shows that they present genetic material that it can be used in breeding programs that have the purpose of producing hybrid for silk production.

Este trabalho teve como objetivo avaliar o polimorfismo em lagartas do bicho-da-seda de diferentes linhagens utilizando a eletroforese de isoenzimas para detectar marcadores bioquímicos e investigar a genética de populações para essas linhagens. Foram utilizados como amostras extratos individuais de glândulas sericígenas de lagartas do segundo dia da quinta idade, de sete linhagens japonesas e oito linhagens chinesas puras mantidas pela Cocamar-Cooperativa Agroindustrial de Maringá. As isozimas fosfatase ácida (ACP), fosfatase alcalina (AKP) e anidrase carbônica (CA) foram avaliadas por meio de eletroforese em géis de amido de milho a 14%. As esterases (EST) foram analisadas por meio de eletroforese vertical em géis de poliacrilamida a 10% e coloração com α e ß- naftil acetato. Foram observados 21 locos, dentre os quais quatro (19.05%) são polimórficos, Est-11, Acp, Akp, Ca. O índice de fixação (Fis) para as isozimas analisadas foi 0.0751, indicando excesso de homozigotos. O valor de Fst (0.6128) permite sugerir que as linhagens estão bem diferenciadas. O dendograma obtido a partir dos valores de distância genética não separou totalmente as linhagens chinesas e japonesas analisadas. Essa avaliação preliminar das linhagens de B. mori mostra que elas apresentam material genético que pode ser utilizado em programas de cruzamentos que tenham a finalidade de produzir híbridos para produção de seda.

Characterization of differentially expressed stress-associated proteins in starch granule development under heat stress in wheat (Triticum aestivum L.).

Abiotic stress causes abrupt increase in the expression of stress-associated proteins, which provide tolerance by modulating the defense mechanism of plants. Small heat shock proteins (sHSPs) and anti-oxidant enzymes are important for environmental stress tolerance of the plants. In this study, two full-length cDNAs encoding small heat shock protein (sHSP) and superoxide dismutase (SOD), designated as TasHSP and SODI were identified and characterized from C-306 (thermotolerant) and PBW343 (thermosusceptible) cultivars of wheat (Triticum aestivum L.). An alpha crystalline domain was observed in TasHSP and manganese/iron binding domain in case of SODI. Quantitative real-time PCR showed very high transcript level of TasHSP and SOD in C-306 compared to PBW343 at different stages of growth and against differential heat stress (HS). Under differential HS at milky-dough stage, the fold change in transcript of both TasHSP and SOD was observed maximum in C-306, compared to PBW343. Protein profiling and isoenzymes analysis showed the expression of several heat-stable proteins and prominent isoenzymes of SOD in C-306, compared to PBW343. Scanning electron microscopy (SEM) of starch granules showed globular, well-shaped and more numbers of endospermic cells in C-306, compared to defragmented, irregular shaped and shrunken granules in case of PBW343 under HS treatment (42°C for 2 h). Diurnal change in soluble starch synthase (SSS) activity showed an increase in the activity during afternoon (35°C), compared to morning (29°C) and evening (32°C) in both the cultivars. Under heat stress (42°C for 2 h), a drastic decrease in the SSS activity was observed, due to the thermal denaturation of the enzyme. Thermotolerance capacity analyzed using cell membrane stability (CMS) showed significantly higher CMS in case of C-306, compared to PBW343 at different stages of growth. Findings suggest that abundance of TasHSP and SODI during milky-dough stage plays a very important role in starch granule biosynthesis. The mechanism may be further exploited to develop tolerant wheat cultivar with high quality seeds.

Comparative electrophoretic profile of proteins and esterases in healthy silkworm larvae (Bombyx mori Lineu, 1758) and infected with nucleopolyhedrovirus / Perfil eletroforético comparativo de proteínas e esterases em lagartas de bicho da seda (Bombyx mori Lineu, 1758) saudáveis e infectadas por nucleopoliedrovírus

Total proteins and esterases from silk gland extracts of Bombyx mori silkworm were characterized and compared with electrophoretic profiles of prepared extracts with silkworm glands infected with nucleopolyhedrovirus (BmNPV). SDS-PAGE (7%) gels was used for the total proteins, and 10% native PAGE for esterases. In the silk glands extracts of healthy silkworms, it was observed seven protein zones with molecular weight varying between 10 kDa (P1) and 60 kDa (P7). In the infected silkworms, a new zone named P8 (90 kDa) was also detected. Esterases activity at 5th instar larvae underwent changes after the infection with BmNPV, since there was a reduction (EST-6 and EST-7) and an increase (EST-8) in the intensity of the regions of esterases activity, and specificity of EST-9 to ß-naphthyl acetate. Those alterations observed in the expression of genes after the infection with the nucleoplyhedrovirus can be used as markers to detect infections in B. mori.

Nesse estudo foram identificadas e comparadas as alterações das proteínas totais e esterases em extratos de glândula sericígena de lagartas de Bombyx mori sadias e infectadas por nucleopoliedrovírus (BmNPV), empregando eletroforese SDS-PAGE com géis a 7% para proteínas totais, e PAGE a 10% para esterases. Nos extratos de glândulas sericígenas de lagartas saudáveis, foram observadas sete regiões proteicas com peso molecular que variam entre 10 kDa (P1) a 60 kDa (P7). Naquelas infectadas pelo BmNPV, uma nova região denominada de P8 (90 kDa) foi detectada. A atividade das esterases no 5º instar larval sofreu alterações após a infecção pelo BmNPV porque houve redução (EST-6 e EST-7) e aumento (EST-8) na intensidade das regiões de atividade de esterases; e especificidade da EST-9 com o substrato ß-naftil acetato. Essas alterações na expressão gênica, após a infecção pelo nucleopoliedrovírus, poderão ser utilizadas como marcadores para detecção da infecção em B. mori.

Valor pronóstico de las isoenzimas de LDH en pacientes con linforma no hodgkin / Prognostic value of LDH isoenzymes in patients with non-Hodgkin linforma

La enzima lactato deshidrogenasa (LDH) es un factor pronóstico en Linfoma No Hodgkin (LNH). El objetivo del trabajo consistió en evaluar prospectivamente el valor pronóstico de las isoenzimas de LDH en pacientes con LNH. Se estudiaron 67 pacientes de primera consulta con diagnóstico de LNH, sin tratamiento previo, VIH negativo y sin otras enfermedades, tiempo promedio de seguimiento 30 meses (rango 3-48 meses). Las muestras de suero se recolectaron previas al tratamiento. La LDH total (LDHT) e isoenzimas de LDH se determinaron respectivamente por método cinético y electroforesis de proteínas en gel de agarosa. Se procesaron muestras de 122 controles sanos para establecer los valores de referencia de las isoenzimas de LDH. 49(73%) LNH agresivos y 18(27%) LNH indolentes y según el Índice Pronóstico Internacional (IPI), 60 (90%) bajo riesgo y 7(10%) alto riesgo. Las isoenzimas LDH1, LDH2, LDH3, LDH4 y LDH5 presentaron niveles absolutos significativamente elevados en 25 (37%), 29 (43%), 32 (48%), 20 (39%) y 11 (16%) de los casos respectivamente (p<0,0001). La actividad porcentual de LDH4 en los pacientes con LNH agresivos fue significativamente superior respecto al grupo de LNH indolentes (p=0,01). En el análisis univariado, valores absolutos elevados de LDH1 se asociaron significativamente con una sobrevida global disminuida (p=0,0064) en el grupo total de pacientes. LDH1 conservó su valor pronóstico aún en el grupo de pacientes con valores normales de LDHT (p=0,04). En pacientes con LNH agresivos, valores elevados de LDHT e IPI alto riesgo se asociaron significativamente con una menor sobrevida global (p<0,05). En el análisis multivariado la LDHT e IPI resultaron factores pronósticos independientes de la sobrevida. Alteraciones específicas del patrón de isoenzimas de LDH sugieren la relación de LDH4 con la biología del tumor y su actividad proliferativa en LNH agresivos y el valor pronóstico de LDH1 como factor adverso de la sobrevida en el análisis univariado.

Lactate dehydrogenase (LDH) is a prognostic factor in non-Hodgkin lymphoma (NHL). Our objective was to evaluate prospectively the prognostic value of LDH isoenzymes in patients with NHL. We studied 67 newly diagnosed NHL patients, previously untreated, HIV-negative and free from other disease, median follow-up of 30 month (range 3-48 month). Before starting treatment serum samples were collected for the determination of total LDH (LDHT) and LDH isoenzymes that were respectively assayed by kinetic method and protein electrophoresis in agarose gel. In order to set reference values of LDH isoenzymes samples from122 healthy controls were processed. Results: 49(73%) of the patients were aggressive NHL and 18(27%) indolent NHL and according to the International Prognostic Index (IPI), 60(90 %) low risk and 7(10%) high risk. High absolute values of LDH1, LDH2, LDH3, LDH4 and LDH5 isoenzymes were significantly elevated in 25 (37%), 29 (43%), 32 (48%), 20 (39%) and 11 (16%) of cases respectively (p<0,0001). The percentage value of LDH4 activity in aggressive NHL patients was significantly higher compared to indolent NHL group (p=0,01). In univariate analysis increased LDH1 absolute values were significantly associated with decreased overall survival in the total group of patients (p = 0.0064). LDH1 remained a prognostic factor for survival even when considering the group of patients with normal serum LDHT values (p = 0.04). In patients with aggressive NHL increased values of LDHT and high risk IPI were significantly associated with decreased overall survival (p<0.05). In a multivariate analysis LDHT and IPI score were independent prognostic factor for survival.