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Aim: To examine whether or not non-secretion of ABH substances and non-tasting of PTC are risk factors exhibiting positive interactions for tuberculosis. Methodology: A total of 210 individuals comprising 110 tuberculosis patients (test group) and 100 apparently healthy subjects (control group) participated in this study. Secretors and non-secretors were determined among the study participants by haemagglutination inhibition test and Tasters and non-tasters were determined using phenylthiocarbamide (PTC) taste strips (0.0143 mg/strip). Results: Of the 110 tuberculosis patients, 65 (59.1%) and 45 (40.9%) were secretors and non-secretors respectively while 49 (44.5%) were tasters and 61 (55.5%) were non-tasters. Of the 100 control subjects, 78% and 22% were secretors and non-secretors respectively while 67% and 33% were tasters and non-tasters respectively. Non-secretors of ABH substances were significantly more associated with test patients than controls (χ2 = 8.62, df = 1, p = 0.002). Non-tasters of PTC were significantly more associated with test patients than controls (χ2=10.68, df=1, p=0.001). When combined, secretors and tasters were significantly lower in the test group than in the control group (χ2=13.44, df=1, p<0.001) while non-secretors and non-tasters were significantly higher in the test group than in the control group (χ2=9.77, df=1, p=0.002). Individuals who were both non-secretors and non-tasters were significantly associated with tuberculosis compared to those who were not (OR 3.5; 95% C.I 1.59-7.51). Conclusion: This study shows that there is a remarkable increased incidence of tuberculosis in individuals who are both unable to secrete ABH substances and taste PTC.
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Objective To prepare 99Tcm-human epidermal growth factor receptor 2 (HER2) affibody (ABH2) and explore its feasibility as an imaging agent in HER2-positive breast cancer.Methods Sodium glucoheptonate and SnCl2 · 2H2O were used to label ABH2 with 99Tcm.The labeling yield and radiochemical purity of 99Tcm-ABH2 were determined.The stability of 99Tcm-ABH2 was tested in PBS and serum.The equilibrium disassociation constant (Kd) of 99Tcm-ABH2 was measured with MBA-MD-361 breast cancer cells.SPECT/CT imaging was carried out at 1.0 h and 4.5 h after injection of 37 MBq 99Tcm-ABH2 in nude mice (n=4) xenografted with MBA-MD-361 breast cancer.The T/NT (liver,brain,lung,heart,bone and muscle) ratios were deduced from SPECT/CT acquired data.For the blocking experiment,200 μg ABH2 was injected intravenously before 99Tcm-ABH2 injection.SPECT/CT imaging was performed in the same way.The T/NT ratios were compared between non-blocked and blocked groups by one-way analysis of variance.Results Fhe labeling yield of 99Tcm-ABH2 was above 99％.99Tcm-ABH2 was substantially stable in PBS and serum;the radiochemical purity was (95.0± 1.0)％ after incubation with serum at 37 ℃ for 6.0 h.The Kd of 99Tcm-ABH2 was 1.7 nmol/L.The radioactive uptake in cancer was visualized at 1.0 h and 4.5 h after injection of 99Tcm-ABH2 in HER2 positive MDA-MB-361 breast cancer.99Tcm-ABH2 was cleared out mainly through urinary system.The ratios of tumor to liver,lung,brain,heart,muscle and bone were 1.81± 0.60,8.95±1.13,20.08±6.12,7.61±0.56,10.62±1.78,11.42±2.07,respectively at 4.5 h after 99Tcm-ABH2 injection.After ABH2 blocking,the corresponding T/NT ratios were 0.60±0.23,3.05± 1.38,5.24±2.17,2.42±1.02,8.16±2.66,2.76±0.48 (F=29.38,P＜0.05) respectively.Conclusion 99Tcm-ABH2 could be synthesized with high purity,and this new agent could be used to image HER2-positive breast cancer specifically.
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Objective To detect amniotic fluid ABH blood group substances and ABO blood group genotype by the polymerase chain reaction with sequence-specific primers(PCR-SSP) to increase the prenatal diagnosis of fetal ABO blood group .Methods 53 pregnant women with gestational age 16 -25 weeks were selected .Amniotic fluid was extracted for detecting ABH blood group substances by the serological indirect agglutinating reaction ;the amniotic fluid cells were separated for extracting DNA .Then the PCR-SSP technique was adopted to analyze the ABO blood group genotypes .Results 16 specimens of amniotic fluid were non-se-creting type phenotype(30 .2% ) and 37 specimens of amniotic fluid were secreting type phenotype (69 .8% );48 specimens of amni-otic fluid were detected out the ABO blood group genotype by the PCR-SSP method .ABO blood group of fetal amniotic fluid cells by the gene identification was consistent to the detection results of amniotic fluid secreting type ABH blood group substances .Con-clusion The PCR-SSP technique can accurately detect the fetal amniotic fluid cells ABO blood group .
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In order to isolate novel organic solvent-tolerant (OST) lipases, a metagenomic library was built using DNA derived from a temperate forest soil sample. A two-step activity-based screening allowed the isolation of a lipolytic clone active in the presence of organic solvents. Sequencing of the plasmid pRBest recovered from the positive clone revealed the presence of a putative lipase/esterase encoding gene. The deduced amino acid sequence (RBest1) contains the conserved lipolytic enzyme signature and is related to the previously described OST lipase from Lysinibacillus sphaericus 205y, which is the sole studied prokaryotic enzyme belonging to the 4.4 a/ß hydrolase subgroup (abH04.04). Both in vivo and in vitro studies of the substrate specificity of RBest1, using triacylglycerols or nitrophenyl-esters, respectively, revealed that the enzyme is highly specific for butyrate (C4) compounds, behaving as an esterase rather than a lipase. The RBest1 esterase was purified and biochemically characterized. The optimal esterase activity was observed at pH 6.5 and at temperatures ranging from 38 to 45 °C. Enzymatic activity, determined by hydrolysis of p-nitrophenyl esters, was found to be affected by the presence of different miscible and non-miscible organic solvents, and salts. Noteworthy, RBest1 remains significantly active at high ionic strength. These findings suggest that RBest1 possesses the ability of OST enzymes to molecular adaptation in the presence of organic compounds and resistance of halophilic proteins.
Con el fin de aislar nuevas variantes de lipasas tolerantes a solventes organicos (OST), se construyo una libreria metagenomica a partir de ADN obtenido de una muestra de suelo de bosque templado. A traves de un monitoreo en dos etapas, basado en la deteccion de actividades, se aislo un clon con actividad lipolitica en presencia de solventes organicos. La secuenciacion del plasmido pRBest recuperado del clon positivo revelo la presencia de un gen codificante de una hipotetica lipasa/esterasa. La secuencia deducida de amino acidos (RBest1) contiene los motivos conservados de enzimas lipoliticas y esta relacionada con la lipasa OST previamente descrita de Lysinibacillus sphaericus 205y, que es la unica enzima procariota estudiada perteneciente al subgrupo 4.4 de a/ß hidrolasas (abH4.04). Estudios in vivo e in vitro sobre la especificidad de sustratos de RBest1, utilizando triacil-gliceroles o p-nitrofenil-esteres, respectivamente, revelaron que la enzima es altamente especifica para compuestos butiricos (C4), comportandose como una esterasa y no como una lipasa. La esterasa RBest1 fue purificada y caracterizada bioquimicamente. La actividad optima de esterasa fue observada a pH 6,5 y las temperaturas optimas fueron entre 38 y 45 °C. Se establecio que la actividad enzimatica, determinada por hidrolisis de p-nitrofenil esteres, es afectada en presencia de diferentes solventes organicos miscibles y no miscibles, y tambien sales. Notoriamente, RBest1 permanece significativamente activa a elevadas fuerzas ionicas. Estos hallazgos sugieren que RBest1 posee la capacidad de las enzimas OST de la adaptacion molecular en presencia de compuestos organicos, asi como la resistencia de las proteinas halofilas.
Subject(s)
Esterases/isolation & purification , Lipase/isolation & purification , Metagenomics , Amino Acid Sequence , Bacillaceae/enzymology , Bacterial Proteins/chemistry , Butyrates/metabolism , Conserved Sequence , DNA , Esterases/classification , Germany , Hydrogen-Ion Concentration , Hydrolysis , Lipolysis , Lipase/classification , Molecular Sequence Data , Osmolar Concentration , Phylogeny , Recombinant Proteins/metabolism , Sequence Alignment , Sequence Homology, Amino Acid , Soil Microbiology , Substrate Specificity , Salts/pharmacology , Solvents/pharmacology , Temperature , Trees , Triglycerides/metabolismABSTRACT
Objective To detect the difference of intensity of A ,B ,H antigen on erythrocytes between neonates and adult .Meth-ods Anti-A ,anti-B and anti-H serum were diluted in multiple proportions ,then add the sample erythrocytes into the test tube to react with the serum above .Observe the agglutination between the erythrocytes and the serum ,and score for every agglutination re-action .The total of the score in different dilution of every sample were brought into statistics analysis by SPSS software .Results The A ,B ,H antigen intensity on the surface of erythrocyte of neonates were less than that of adult (P0 .05) .Conclusion There is a significant difference of A ,B ,H antigen intensity on erythrocytes between neonates and adult .It also can be concluded that the varity of the antigen has effect on intensity of A ,B ,H antigen in erythrocytes ,except sex .
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En investigaciones previas se identificaron antígenos ABH en ejemplares adultos de Ascaris lumbricoides. Los parásitos sólo presentaron epitopes expresados en sus respectivos hospedadores, adquiridos por absorción. Como el parásito adulto vive en el intestino delgado, podría presentarlos debido a que la larva de cuarto estadio conserva los epitopes absorbidos en la migración y los mantiene durante el proceso madurativo a adulto o bien el ejemplar adulto los adquiriría de las secreciones intestinales. El objetivo fue estudiar la absorción de epitopes solubles ABH por larvas mantenidas en cultivo. Las larvas fueron obtenidas de la eclosión de huevos y se recolectaron y cultivaron en medio RPMI. Se adicionó antígeno soluble B en tres tubos que fueron incubados 2, 3 y 6 días y antígeno soluble A en dos tubos cultivados 6 días. Cada tiempo de cultivo tuvo su control negativo de absorción, donde se investigaron productos de excreción-secreción (ES) Tipo antígenos ABH. La técnica de Inhibición de la Aglutinación Semicuantitativa demostró epitopes B en las larvas cultivadas 3 días y en las incubadas 6 días epitopes A y B. Solamente se evidenciaron productos ES símil A y B en el Control de 6 días. Se concluye que A. lumbricoides puede absorber determinantes ABH a partir de antígenos solubles.
Previous experiences have identified ABH antigens in A. lumbricoides's adult specimens. The parasites only showed epithopes expressed in their respective hosts. It was demonstrated that epithopes are acquired by absorption by means of the culture of larvae with erythrocytes. As the adult parasite lives in the small intestine, it could be present there because the fourth stage larva retains the epithopes absorbed in the migration and keeps them during the maturation process to adult, or the adult specimen would acquire them from intestinal secretions. Larvae were obtained by hatching of eggs and they were collected and cultured in RPMI medium. The aim of this work was to study the ABH soluble epithopes by larvae in culture. B soluble antigen was added into three tubes, which were incubated for 2, 3 and 6 days, and A soluble antigen was poured into two tubes, both cultured for 6 days. Each culture time had its absorption Negative Control where ABH like antigen excretion/secretion products (ES) were investigated. The Semiquantitative Inhibition Agglutination Test showed B epithopes in the larvae cultured for 3 days and A and B epithopes in the larvae incubated for 6 days. Similar A and B ES products were only evidenced in the Control of 6 days. The results conclude that A. lumbricoides may absorb ABH antigenic determiners from soluble antigens.
Subject(s)
Absorption , Allergy and Immunology , Antigens , Ascaris lumbricoides , Parasites/parasitologyABSTRACT
A cross sectional study was done with 42 apparently healthy persons aged 6 years and above from both sexes. Most of them are blood donors in the department of Transfusion Medicine, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh. Few, other than blood donor, were selected from the same locality. Five ml venous blood was collected with all aseptic precautions. ABO blood grouping and Lewis phenotyping were done by tube method. ABO reverse grouping was also done from serum. With all precautions 2 ml of saliva was collected from all subjects. Secretor status was detected from the saliva by haemagglutination inhibition method. ABO blood grouping shows 36% 'O' group, 24% 'A' group, 33% 'B' group and 7% 'AB' group. Distribution of Lewis phenotype are Le(a+b-) 19%, Le(a-b+) 53%, Le(a-b-) 26% and Le(a+b+) 2% only. 60% of study population was ABH secretor and 40% non-secretor.
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Los antígenos ABH, productos de la interacción de 2 sistemas genéticos, Hh y ABO, están sujetos a leyes de herencia y pueden estar localizados no sólo en los eritrocitos, sino también en la mayoría de las células humanas. El objetivo del este trabajo fue investigar la expresión de antígenos ABH en pacientes con lesiones orales premalignas y malignas orales. Se trabajó con muestras incluidas en tacos de parafina de pacientes con lesiones orales (n= 57). Los pacientes fueron clasificados en 2 grupos: a) lesiones premalignas y malignas diagnosticadas clínica y anatopatológicamente y b) lesiones benignas (n=93). Se investigaron los antígenos ABH por la técnica de inmunoadherencia específica modificada. Se utilizó la adherencia al tejido vascular como control positivo y al tejido adiposo como control negativo. Los resultados fueron semicuantificados desde adherencia fuertemente positiva a negativa. Se observó una significativa relación entre la expresión antigénica ABH y el grado de malignidad de las lesiones analizadas (P Yates= 0,005). La pérdida de reactividad ABH en los sitios de mayor invasividad tumoral se correlaciona con el grado del desarrollo del tumor, el grado histológico y su malignidad.
The ABH antigens, which are produced by the interaction of 2 genetic systems, Hh and ABO, are subjected to laws of heredity and may be located not only in the erythrocytes, but also in most of the human cells. The objective of this paper was to investigate the expression of ABH antigens in patients with premalignant and malignant oral lesions. Work was done with samples included in paraffin plugs in patients with oral lesions (n= 57). The patients were classified into 2 groups: a) clinical and anatomopathologically diagnosed premalignant and malignant lesions, and b) benign lesions (n=93). The ABH antigens were investigated by the modified specific immunoadherence technique. Adherence to the vascular tissue was used as a positive control, whereas adherence to the fat tissue was considered as a negative control. The results were semiquantified from strongly positive to negative adherence. A significant relation between the ABH antigen expression and the degree of malignancy of the analyzed lesions (P Yates= 0.005) was observed. The loss of ABH reactivity in the sites of greater tumoral invasiveness is correlated with the tumor development degree, the histological degree and its malignancy.
Subject(s)
Humans , Antigens , Mouth NeoplasmsABSTRACT
The association of the ABO antigen-antibody titres with the ABH secretor status in duodenal ulcers was examined in a sample of 196 patients and 182 healthy controls. The risk for group A and group B patients depended upon the strengths of the antigens and the ABH secretor status. Further the risk was high for low antibody titres in non-secretor patients than in secretor patients. Increased demand on the antibodies for mucoprotection in the absence of the A and B antigens in the gastro-intestinal mucosa or insufficient production of antibody by the lymphocytes could be the possible reasons.
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One step dot-ELISA method for the rapid determination of ABO blood groups in humam body fluids(or stains)was established using enzyme-labelled anti-A,--B and anti--H monoclonal antibody (McAb).The sample was applied on the nitrocellulose membrance as a dot. After washing, the appropriate McAb wasadded on top of the dot, and then followed by adding 3, 3'-diaminobenzidine(DAB). The brown color indi-cated the positive reaction. The ABO blood typing of 521 saliva samples including both secretor and non-se-cretor were carried out. All the results were correct. The advantages of this method are accurate, sensitive,rapid, easy to perform, as well as not time consuming. It is more sensitive than the conventional hemaggluti-naton inhibition test.
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The distribution of ABH substances in human tissue cells was studied using the specific red cell adherence test(SRCA test).Tissues were taken from 11 cadavers of known ABO type and secrete status,fixed with 10% neutral formalin,Isolated abdominal skins kept in room temperature for 1～13 days were also observed for the purpose of studing the influence of the time elapsed after death on SRCA test results ABH substances were found in mucous membranes,mucous glands and prostate glands.ABH substances in those tissues were controlled by secrete status.ABH substances were also found in endothelia of blood vessels,stra- tified epithelia,acinar cells of pancreas and sweat glands.We firstly found that ABH substances were present in epithelia of pulmonary alveoli and epi- thelia of small bile duct in liver. Using SRCA test,the ABO blood type were correctly demonstrated in 11 isolated abdominal skins kept in room temperature for 1～13 days.
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Rhesus monkey (Macaca mulatta)ABO blood typing was carried out by agglutination test with anti human sera, and monoclonal antibodies against A and B blood group antigcus Blood Samples of 113 monkeys were tested. The results showed that there wete no human like A and B agglutinins and anti-A and anti-B agglutnins on the surface of the red blood cells and in the sera of monkeys. How ever human like blood group substances were found in monkey's galiva lemonsliated by agglutination inhibition test. The ABO phenotype frequoncies of Rhesus monkey were as follows: A-17. 70 %;B-52,21 %;AW-20, 36% and O-9. 73 % which of ther from foreign reports.