ABSTRACT
@#Abstract: Objective To analyze the drug sensitivity and the carrying of carbapenem resistant gene of Acinetobacter baumannii isolated from clinical patients and clinical objects, and analyze the homology of strains to provide support for the control of nosocomial infection. Methods A total of 38 strains of Acinetobacter baumannii isolated from patients and clinical objects surface were collected from January 2019 to August 2020. The antimicrobial susceptibility was tested by the minimum inhibitory concentration method. In addition, the resistance related genes were detected by polymerase chain reaction method, and homology analysis was performed by enterobacterial repetitive Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR). Results All 34 strains of Acinetobacter baumannii isolated from Clinical patients and 4 strains isolated from clinical objects carried blaOXA-51 and imp resistance genes, neither of them carried blaVIM gene. 32 Acinetobacter baumannii carrying blaOXA-23 gene, 28 strains carrying blaTEM gene, 7 strains carrying blaOXA-58 gene. After cluster analysis, 38 Acinetobacter baumannii isolates were classified into 7 genotypes (expressed A, B, C, D, E, F, G), and cluster E and cluster G were the main clusters, containing 12 strains (12/38, 31.6%) and 18 strains (18/38, 47.4%), respectively, as the main prevalent clonal strains. Conclusions Acinetobacter baumannii isolated from different sources have the significant differences in drug resistance and carry different resistance genes. There is no direct correlation between patients and environmental isolates of Acinetobacter baumannii belonging to different clonal strains. Also, there aren’t significant correlation between clinical patients infected with Acinetobacter baumannii.
ABSTRACT
OBJECTIVE: To study the effect of stains on the purity detection results of human serum albumin by agarose gel electrophoresis, thus to select appropriate stain so that the detection result of agarose gel electrophoresis method can be basically consistent with the method specified in the 2015 edition of Chinese Pharmacopoeia, cellulose acetate membrane electrophoresis method. METHODS: Acid blue, amino black and ponceau were used in agarose gel electrophoresis to detect the purity of 30 batches of human serum albumin samples, and the results were compared with those obtained by cellulose acetate membrane electrophoresis. RESULTS: Impurity bands could not be detected by ponceau stain; the results of amino black staining was significantly lower than that of cellulose acetate membrane electrophoresis. There was no statistical difference between the results of amino black staining and cellulose acetate membrane electrophoresis. The results of 25 batches of samples detected by cellulose acetate membrane electrophoresis were within the 95% confidence interval of the results detected by amino black staining. CONCLUSION: The category of stain has great influence on the detected purity of human serum albumin when using agarose gel electrophoresis. The results of amino black staining is consistent with those of the method recorded in the Chinese Pharmacopoeia in use.
ABSTRACT
Caprine arthritis encephalitis causes considerable losses in goat production. The main form of the caprine arthritis encephalitis virus transmission is through the ingestion of colostrum or milk from infected females. However, some transmissions cannot be explained in this manner. Therefore, this study aimed to evaluate transplacental transmission of caprine arthritis encephalitis virus. Blood samples were collected from 283 newborn kids of Anglo-Nubian and Saanen breeds born from seropositive and seronegative goats. Samples were collected immediately after birth and analyzed with agarose gel immunodiffusion and western blot. All samples were negative in the agarose gel immunodiffusion. However, the western blot test demonstrated that four kids were born positive for caprine arthritis encephalitis virus. This result indicates that although in a low frequency (1.4%), there is a possibility of transplacental transmission of small ruminant lentivirus.(AU)
A artrite encefalite caprina causa perdas consideráveis para a produção caprina. A principal forma de transmissão do vírus da artrite encefalite caprina é a ingestão de colostro ou leite de fêmeas infectadas. No entanto, algumas transmissões não podem ser explicadas por esta via. Dessa forma, este estudo teve como objetivo avaliar a transmissão do vírus da artrite encefalite caprina por via transplacentária (vertical). Foram realizadas coletas de sangue em 283 crias recém-nascidas das raças Anglo-Nubiana e Saanen, provenientes de progenitores soropositivos e soronegativos. As amostras foram coletadas logo após o nascimento e analisadas pelas técnicas de imunodifusão em gel de agarose e western blot. No teste de imunodifusão em gel de agarose, nenhum cabrito foi detectado reagente. Porém, no teste de western blot, quatro cabritos nasceram soropositivos. Esse resultado indica que, apesar de baixa frequência (1,4%), existe a possibilidade de transmissão via transplacentária do lentivírus de pequenos ruminantes.(AU)
Subject(s)
Animals , Infant, Newborn , Ruminants , Lentiviruses, Ovine-Caprine , Arthritis-Encephalitis Virus, Caprine , Infectious Disease Transmission, Vertical , Livestock Industry , Animals, Newborn/virologyABSTRACT
Objective To explore the influence of lower concentration of cell free fetal fraction DNA in maternal plasma on non-invasive prenatal test(NIPT) .Methods A total of 3240 pregnant women accepted NIPT in Foshan Maternal and Children′s Hos-pital from April ,2015 to March ,2016 were analyzed retrospectively ,and 150 samples of which were male fetus judged by Z score of Y chromosome and the cell free fetal fraction DNA were lower than 8% were selected .The cell free fetal fraction DNA were in-creased by agarose gel electrophoresis ,then conducted NIPT ,compared with the results of aneuploidy screening .Results The cell free fetal fraction DNA were increased from 5% to 9 .2% by agarose gel electrophoresis .The result of NIPT after increasing fetal fraction was consistent with it before .Conclusion Concentration of cell free fetal fraction DNA has no influence on the result of NIPT when cell free fetal fraction DNA is above 5% .
ABSTRACT
Objective To explore the influence of lower concentration of cell free fetal fraction DNA in maternal plasma on non-invasive prenatal test(NIPT) .Methods A total of 3240 pregnant women accepted NIPT in Foshan Maternal and Children′s Hos-pital from April ,2015 to March ,2016 were analyzed retrospectively ,and 150 samples of which were male fetus judged by Z score of Y chromosome and the cell free fetal fraction DNA were lower than 8% were selected .The cell free fetal fraction DNA were in-creased by agarose gel electrophoresis ,then conducted NIPT ,compared with the results of aneuploidy screening .Results The cell free fetal fraction DNA were increased from 5% to 9 .2% by agarose gel electrophoresis .The result of NIPT after increasing fetal fraction was consistent with it before .Conclusion Concentration of cell free fetal fraction DNA has no influence on the result of NIPT when cell free fetal fraction DNA is above 5% .
ABSTRACT
Background: Oxygen transport is altered in hemoglobinopathies. Aim: To study the distribution of hemoglobinopathies in Andean subjects without African ancestry. Material and Methods: We analyzed blood samples of 1,407 subjects aged 18 to 59 years (58% females), living in the central Andean region of Colombia, referred to discard hemoglobinopathies. The frequency and type of hemoglobinopathy was established by capillary and agarose gel electrophoresis. Results: The frequency of hemoglobinopathies was 34.5% and higher among females. The structural variants found were: AS-heterozygous hemoglobin (8.1%), homozygous SS (3.7%), heterozygous SC (2.2%), AC heterozygotes (0.5%) and heterozygous AE (0.3%). Quantitative variants found were Hb A-Beta thalassemia (13.91%) and Hb H (0.06%), Beta-thalassemia heterozygotes C (0.88%), S-Beta thalassemia heterozygotes (6.07%) and compound heterozygous SC/Beta thalassemia (0.25%), with a persistence of fetal hemoglobin 0. Composite thalassemia was also found in 31%. All techniques showed good correlation and capillary electrophoresis demonstrated a greater detection of hemoglobin variants. Conclusions: The frequency of hemoglobin variants in the analyzed population was high, which is an important public health indicator. The most common hemoglobin variant was HbA/Increased structural Hb A2 and the mos frequent structural hemoglobinopathy was sickle cell trait. Capillary electrophoresis can discern any Hb variants present in the population.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Hemoglobinopathies/epidemiology , Hemoglobins/analysis , Colombia/epidemiology , Electrophoresis, Agar Gel , Electrophoresis, Capillary , Hemoglobinopathies/classification , Hemoglobinopathies/diagnosis , Hemoglobinopathies/ethnology , Retrospective StudiesABSTRACT
Objective To investigate the detection rates and types of hemoglobinopathy in Xiaolan district of Zhongshan city to provide the basis for prepotency .Methods 7 652 pregnant women with pregnancy detection in our hospital from June 2010 to De‐cember 2013 were selected and performed the electrophoresis by the Hydrasys automatic hemoglobin electrophoresis system .The hemoglobin components were scanned out and performed the statistical analysis .Results Among 7 652 samples ,1 057 cases of sus‐pected hemoglobinopathy were screened out with the detection rate of 10 .36% .Among them ,511 cases(6 .68% ) were Hb A2 3 .5% (suspected β‐thalassemia) ,the detection rate of Hb A2 >3 .5% was lower than that of Hb A2 < 2 .0% ,the difference had statistical significance(P< 0 .05) .11 cases (0 .14% ) were Hb C or E ,9 cases (0 .12% ) were Hb J or K ,8 cases (0 .10% ) were Hb H ,7 cases (0 .09% ) were Hb Bart′s ,1 case was Hb CS ,2 cases were Hb G and 1 case was Hb D ,which accounted for 0 .04% .Conclusion Hemoglobin electrophoresis can screen hemoglobinopa‐thy effectively .The hemoglobinopathy screening in this area should be conducted as soon as possible ,which has an important signif‐icance to prepotency .
ABSTRACT
Background: The detection and analysis of microsatellites is very important for the mapping of genetic diseases because they are commonly used as genetic markers. Microsatellite marker D19S884 has been associated with polycystic ovary syndrome (PCOS), the most common reproductive endocrine disease of women in their childbearing years. It is responsible for an estimated 70% of cases of anovulatory infertility. In this work, we detected microsatellites in DNA extracted from the blood of PCOS patients. Methods: DNA microsatellites were amplified by polymerase chain reaction (PCR) using a pair of specific primers tagged with fluorescence to yield products of 160–200 base pairs in length. Alleles were separated on 4% low-melting agarose gels; stained with a safe gel staining, GelRed™, which is an alternative to ethidium bromide; and visualised by ultraviolet illumination. Results: Bands were observed, but their base-pairs differences were difficult to distinguish. To identify each allele clearly, the PCR products were also analysed using capillary gel electrophoresis for fragment analysis where it was possible to discriminate even in case of difference between two pairs of bases between the alleles. Conclusion: In this article, we present a protocol that combines the use of gel electrophoresis and fragment analysis in the identification of genetic biomarkers for PCOS.
ABSTRACT
Objective To explore the traits of improved polyacrylamide gel electrophoresis (PAGE)-ethidium bromide(EB) staining in the detection of genotype polymorphisms.Methods The methods of PAGE-silver staining,agarose gel electrophoresis (AGE)-EB staining and improved PAGE-EB staining in polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to detect the μ-opioid receptor (OPRM1) A118G genotypes in case group (n =167) infants with intracranial hemorrhage (ICH),and control group(n =163) infants without ICH,to conduct a case study analysis.And the application traits of three methods were compared.Results Genotypes of OPRM1 A118G were GG (169 bp),AG (193 bp,169 bp),AA (193 bp).Both the electrophoresis methods of PAGE-silver staining and PAGE-EB could be used to detect the genotypes of OPRM1 A118G clearly in this study.There was no statistically significant difference between the resolutions of DNA fragments (P =0.884).The first method,which had 13 experiment steps,consuming 4-5 hours,involving 12 kinds of chemical reagents,and the pictures were taken with the camera,was complex,with difficult operation,more time consuming; Compared with the first method,the secondmethod was simple,which had 6 test procedures,consuming 2 hours with 8 reagents,and the pictures were taken by using an automatic gel imager.AGE-EB could not be used to detect genotypes of OPRM1 A118G.Conclusions The method of improved PAGE-EB has the advantages of fast,easy operation,and high resolution,which is worthy of wider application.
ABSTRACT
Investigation of the effects of rhizome atractylodis macrocephalae on yeast prion [PSI+ ]was conducted in this study .Liquid culture containing rhizome atractylodis macrocephalae aqueous extracts was applied to evaluate its therapeutic effects preliminarily .Then yeast replica plating together with Semi Denaturing Detergent Agarose Gel Electrophoresis com-bined with western blot were used to further confirm the results .It's showed that the cure rate of aqueous extracts of rhizome atractylodis macrocephalae on yease prion [PSI+ ]could reach 6% .
ABSTRACT
OBJECTIVE: To synthesize nonviral gene carrier poly(L-glutamic acid)-graft-polypropylenimine(MP-g-PPI) and investigate the capability of the polymers to condense DNA and the stability of MP-g-PPI DNA complexes.
ABSTRACT
Objective To explore the clinical diagnostic value of polyclonal all immunoglobulin.Methods The specimens of pa-tients were simultaneously tested and identified by quantitative immunoglobulins,Immunofixation electrophoresis of serum and u-rine,urine protein electrophoresis,and other ways.Results From 1 patients with rheumatoid arthritis were detected the serum IgG, M,A,KAP and LAM,and urine KAP and LAM,at the same time show the increment of the polyclonal polyclonal all immune glob-ulin hematic disease.Conclusion Polyclonal all immune globulin hematic disease often appear in the complications of chronic in-flammation,which should be paid attention during its in clinical doctors.
ABSTRACT
The antimicrobial efficacy of methylglyoxal (MG) against several gram-negative bacteria including Escherichia coli has been reported. To determine the mechanism of action of MG, molecular interactions between lipid and MG within the liposomal membrane were also investigated. Multilamellar and unilamellar vesicles were prepared from 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). The effect of MG on DPPC liposomal membrane was studied by fluorescence spectroscopy and differential scanning calorimetry. The results indicate that MG interacts mainly with the DPPC head group that produces a significant increase in the fluidity of liposomal vesicles, which could be the cause of a fusion/aggregation effect in microbial cells. The agarose gel electrophoresis study with the genomic DNA extracted from E. coli ATCC 25922 revealed that addition of MG could completely degrade this DNA within 1 h, pointing out to their distinctly high degree of sensitivity towards MG. Further, the drug was able to cross the cell membranes, penetrating into the interior of the cell and interacting with DNA for demonstrating antibacterial activity of MG.
ABSTRACT
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
Subject(s)
Humans , Male , Female , Electrophoresis/methods , Hemoglobinopathies/diagnosis , Diagnostic Techniques and Procedures/standards , Electrophoresis, Agar Gel/methods , Immunodiffusion/methodsABSTRACT
Nosocomial infections in the hospitals disseminated from the cotton fabrics of health care professionals and patients leads to severe complications like respiratory, gastrointestinal and urinary tract infections. Since the hospital based textile materials like nylon and polyester has good surface properties, it can harbour large number of microorganisms. Hence in this study, two different antibacterial drugs showing synergistic properties were attached to different fabricsusing tocopherol acetate as a cross-linker with the aim that, treated fabric could act as barriers against transmission of challenge organisms. Inorder to decrease the drug resistant property of the nosocomial pathogens, a fluoroquinolone and a nitroimidazole compounds were mixed at suitable composition based on their synergistic behaviour. Both the compounds were modified to act as reactive dyes and were covalently bonded to the surface of nylon and polyester in order to impart antibacterial properties. The assay used for measuring antibacterial properties was based on the AATCC Test Method-100. The treated fabric was also subjected to multiple washings to determine its durability based on the AATCC Test Method-124. To determine the mode of action of these drugs, DNA of the drug exposed and unexposed challenge organisms were extracted and analysed by agarose gel electrophoresis. The difference in the number of viable bacteria after ‘0’ contact time and 18 hours contact time with treated fabrics were statistically calculated with P<0.05 considered significant.
ABSTRACT
The wild edible mushroom, Lentinus lepideus has recently been cultivated for commercial use in Korea. While the mushroom has been widely used for nutritional and medicinal purposes, the possible anti-hyperlipidemic action is unclear. The effects of dietary L. lepideus on plasma and feces biochemical and on the liver histological status were investigated in hypercholesterolemic rats. Six-wk-old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Biochemical and histological examinations were performed. A diet containing 5% L. lepideus fruiting bodies reduced plasma total cholesterol, triglyceride, low-density lipoprotein, total lipid, phospholipids, and the ratio of low-density to high-density lipoprotein. Body weight was reduced. The diet did not adversely affect plasma biochemical and enzyme profiles. L. lepideus reduced significantly plasma beta- and pre-beta-lipoprotein, while alpha-lipoprotein content was increased. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red O staining revealed normal findings for mushroom-fed hypercholesterolemic rats. The present study suggests that a diet supplemented with L. lepideus can provide health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.
Subject(s)
Animals , Female , Humans , Rats , Agaricales , Azo Compounds , Body Weight , Cholesterol , Diet , Electrophoresis, Agar Gel , Feces , Fruit , Hepatocytes , Insurance Benefits , Korea , Lentinula , Lipoproteins , Liver , Phospholipids , PlasmaABSTRACT
We investigated diet supplementation with shiitake mushroom fruiting bodies on biochemical and histological changes in hypercholesterolemic rats. Six-wk old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. A diet containing 5% Lentinus edodes fruiting bodies given to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and the LDL/high-density lipoprotein ratio by 34.33, 53.21, 75.00, 34.66, 25.73, and 71.43%, respectively. Feeding mushroom also significantly reduced body weight in hypercholesterolemic rats. However, it had no detrimental effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that L. edodes significantly reduced plasma beta and pre-beta-lipoprotein but increased alpha-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red-O staining showed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that shiitake mushrooms could be recommended as a natural cholesterol lowering substance in the diet.
Subject(s)
Animals , Female , Humans , Rats , Agaricales , Bilirubin , Blood Urea Nitrogen , Body Weight , Calcium , Cholesterol , Creatinine , Diet , Electrophoresis, Agar Gel , Feces , Fruit , Glucose , Hepatocytes , Lentinula , Lipoproteins , Magnesium , Phospholipids , Plasma , Potassium , Serum Albumin , Shiitake Mushrooms , Sodium , Uric AcidABSTRACT
This work was conducted to investigate dietary supplementation of oyster mushroom fruiting bodies on biochemical and histological changes in hyper and normocholesterolemic rats. Six-week old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Feeding a diet containing a 5% powder of Pleurotus ostreatus fruiting bodies to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and LDL/high-density lipoprotein ratio by 30.18, 52.75, 59.62, 34.15, 23.89, and 50%, respectively. Feeding oyster mushrooms also significantly reduced body weight in hypercholesterolemic rats. However, it had no adverse effects on plasma albumin, total bilirubin, direct bilirubin, creatinin, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that P. ostreatus significantly reduced plasma beta and pre-beta-lipoprotein but increased alpha-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red O staining revealed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that a 5% P. ostreatus diet supplement provided health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.
Subject(s)
Animals , Female , Humans , Rats , Agaricales , Azo Compounds , Bilirubin , Blood Urea Nitrogen , Body Weight , Calcium , Cholesterol , Diet , Dietary Supplements , Electrophoresis, Agar Gel , Feces , Fruit , Glucose , Hepatocytes , Insurance Benefits , Lipoproteins , Magnesium , Phospholipids , Plasma , Pleurotus , Potassium , Serum Albumin , Sodium , Uric AcidABSTRACT
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.
Subject(s)
Animals , Male , Female , Albumins , Alpha-Globulins , Beta-Globulins , Crotalus , gamma-Globulins , Refractometry/methods , Blood Protein Electrophoresis/methods , Electrophoresis, Agar Gel/methodsABSTRACT
BACKGROUND: Real-time polymerase chain reaction (PCR) is generally regarded as a very accurate and time-saving method, but it is expensive to run. We evaluated the reliability of an inexpensive and a researcher-friendly gel electrophoresis-based PCR method for the quantification of mRNA, and the results were compared with those obtained by real-time PCR. METHODS: We compared the results of relative quantification for MMP-1 measured by real-time PCR and by ethidium bromide stained-agarose gel electrophoresis after end-point PCR. RESULTS: There was significant but very weak correlation between real-time PCR and end-point PCR for relative quantification of MMP-1 (r=0.16, P<0.01). CONCLUSIONS: Our results suggest that the use of the gel electrophoresis-based end-point PCR is inappropriate for quantifying mRNA. Therefore, in order to confirm the result of relative quantification by end-point PCR, the newly established real-time PCR method or northern hybridization should be applied.