Search

Sarcoptic mange, caused by epidermal infection with Sarcoptes scabiei, negatively impacts the health, welfare, and local abundance of bare-nosed wombats (Vombatus ursinus) in Australia. Improved understanding of the host immune response to disease and its contribution to pathophysiology could be used to inform management actions for this species in and ex situ. To evaluate the immune response of bare-nosed wombats to sarcoptic mange, we validated three assays (haptoglobin, agarose gel electrophoresis, and micro-erythrocyte sedimentation rate) measuring non-specific markers of inflammation using serum samples from free-living wombats from Tasmania (n = 33). We then analysed correlations between the assay results for each non-specific marker of inflammation and wombat's sarcoptic mange scores, and performed histopathological examinations to investigate association of the acute phase response with systemic amyloidosis. We present evidence that haptoglobin and erythrocyte sedimentation rate increased, and albumin decreased, in association with sarcoptic mange scores. This research demonstrates links between the acute phase response and sarcoptic mange severity in bare-nosed wombats, highlighting the utility of non-specific markers of inflammation for aiding assessment of the systemic effects of mange. Showing the value of agarose gel electrophoresis, we also identified specific acute phase proteins warranting future evaluation and found evidence of an immunoglobulin response in mange-affected wombats, revealed by increasing γ-globulins in association with apparent disease severity. Meanwhile, owing to its relatively low resource requirements and rapidity, the erythrocyte sedimentation rate assay may be useful as a point-of-care test to support therapeutic decisions in the field. Our methods and findings are likely to be applicable to a range of other clinical and population health scenarios in captive and free-living wombats, and species impacted by sarcoptic mange globally. [ABSTRACT FROM AUTHOR]

Amyloid-associated neurodegenerative diseases, including Alzheimer's disease (AD), are characterized by the in-brain accumulation of β-sheet structured protein aggregates called amyloids. However, neither a disease model nor therapy is established. We review past data and present new, preliminary data and opinions to help solve this problem. The following is the data-derived model/hypothesis. (1) Amyloid-forming proteins have innate immunity functions implemented by conversion to another sheet conformation, α-sheet. (2) In health, α-sheet structured, amyloid-forming proteins inactivate microbes by co-assembly with microbe α-sheets. Amyloid-forming proteins then undergo α-to-β-sheet conversion. (3) In disease, α-sheet-structured, amyloid-forming proteins over-accumulate and are neuron-toxic. This hypothesis includes formation by virus capsid subunits of α-sheets. In support, we find that 5–10 mM methylene blue (MB) at 54 °C has a hyper-expanding, thinning effect on the phage T4 capsid, as seen by negative stain- and cryo-electron microscopy after initial detection by native gel electrophoresis (AGE). Given the reported mild anti-AD effect of MB, we propose the following corollary hypothesis. (1) Anti-AD MB activity is, at least in part, caused by MB-binding to amyloid α-sheet and (2) MB induces the transition to α-sheet of T4 capsid subunits. We propose using AGE of drug incubated T4 to test for improved anti-AD activity. [ABSTRACT FROM AUTHOR]

Sarcoptic mange, caused by epidermal infection with Sarcoptes scabiei, negatively impacts the health, welfare, and local abundance of bare-nosed wombats (Vombatus ursinus) in Australia. Improved understanding of the host immune response to disease and its contribution to pathophysiology could be used to inform management actions for this species in and ex situ. To evaluate the immune response of bare-nosed wombats to sarcoptic mange, we validated three assays (haptoglobin, agarose gel electrophoresis, and micro-erythrocyte sedimentation rate) measuring non-specific markers of inflammation using serum samples from free-living wombats from Tasmania (n = 33). We then analysed correlations between the assay results for each non-specific marker of inflammation and wombat’s sarcoptic mange scores, and performed histopathological examinations to investigate association of the acute phase response with systemic amyloidosis. We present evidence that haptoglobin and erythrocyte sedimentation rate increased, and albumin decreased, in association with sarcoptic mange scores. This research demonstrates links between the acute phase response and sarcoptic mange severity in bare-nosed wombats, highlighting the utility of non-specific markers of inflammation for aiding assessment of the systemic effects of mange. Showing the value of agarose gel electrophoresis, we also identified specific acute phase proteins warranting future evaluation and found evidence of an immunoglobulin response in mange-affected wombats, revealed by increasing γ-globulins in association with apparent disease severity. Meanwhile, owing to its relatively low resource requirements and rapidity, the erythrocyte sedimentation rate assay may be useful as a point-of-care test to support therapeutic decisions in the field. Our methods and findings are likely to be applicable to a range of other clinical and population health scenarios in captive and free-living wombats, and species impacted by sarcoptic mange globally.

Introduction: Lipoprotein X (Lp‐X) is an abnormal lipoprotein found in multiple disease conditions, including liver dysfunction and cholestasis. High Lp‐X concentrations can interfere with some laboratory testing that may result in spurious results. The detection of Lp‐X can be challenging, and there is currently a lack of consensus regarding the management of Lp‐X other than treating the underlying disease. Case Presentation: A 42‐year‐old female with Hodgkin's lymphoma treated with dexamethasone, high dose cytarabine and cisplatin and vanishing bile duct syndrome confirmed by liver biopsy presented with cholestasis, pseudohyponatremia (sodium, 113 mmol/L; reference range 136–146 mmL/L; serum osmolality, 303 mOsm/kg), and hypercholesterolemia (> 2800 mg/dL, reference range < 200 mg/dL). Lp‐X was confirmed by lipoprotein electrophoresis (EP). Although she did not manifest any specific signs or symptoms, therapeutic plasma exchange (TPE) was initiated based on laboratory findings of extreme hypercholesterolemia, spuriously abnormal serum sodium, and HDL values, and the potential for short‐ and long‐term sequelae such as hyperviscosity syndrome, xanthoma, and neuropathy. During the hospitalization, she was treated with four 1.0 plasma volume TPE over 6 days using 5% albumin for replacement fluid. After the first TPE, total cholesterol (TC) decreased to 383 mg/dL and sodium was measured at 131 mmol/L. The patient was transitioned into outpatient maintenance TPE to eliminate the potential of Lp‐X reappearance while the underlying disease was treated. Serial follow‐up laboratory testing with lipoprotein EP showed the disappearance of Lp‐X after nine TPEs over a 10‐week period. Literature Review: There are seven and four case reports of Lp‐X treated with TPE and lipoprotein apheresis (LA), respectively. While all previous case reports showed a reduction in TC levels, none had monitored the disappearance of Lp‐X after completing a course of therapeutic apheresis. Conclusion: Clinicians should have a heightened suspicion for the presence of abnormal Lp‐X in patients with cholestasis, hypercholesterolemia, and pseudohyponatremia. Once Lp‐X is confirmed by lipoprotein EP, TPE should be initiated to reduce TC level and remove abnormal Lp‐X. Most LA techniques are not expected to be beneficial since Lp‐X lacks apolipoprotein B. Therefore, we suggest that inpatient course of TPE be performed every other day until serum sodium, TC and HDL levels become normalized. Outpatient maintenance TPE may also be considered to keep Lp‐X levels low while the underlying disease is treated. Serum sodium, TC, and HDL levels should be monitored while on maintenance TPE. [ABSTRACT FROM AUTHOR]

Currently, there are several protocols to extract bacterial DNA based on different principles. However, the quantity and the quality of the DNA obtained by each method are highly variable and microorganism dependent. In most of these classical crude methods, highly toxic and hazardous organic solvents such as phenol and chloroform are used for deproteinization, whereas in certain protocols, expensive enzymes including RNases and Proteinases are used. This study was designed to introduce a simple, rapid, inexpensive and effective genomic DNA isolation procedure for Gram-negative bacteria, without the usage of toxic chemicals and costly enzymes. This novel method was compared with another classical method known as the salting-out method, which uses proteinase-K. Concentration and yield of the extracted DNA were determined by gel electrophoresis by comparing the gel band intensity of the sample DNA to that of a DNA quantitation standard and by the Quantus™ fluorometer. According to the results, the yield of extracted DNA was higher in the novel method compared to the salting-out method. Moreover, the entire process was accomplished in less than 2 h with the novel method. Purity and integrity of extracted genomic DNA by both methods were similar. In addition, the quality of DNA was determined using Multicopy Associated Filamentation (MAF) gene amplification by polymerase chain reaction (PCR). Thus, the described technique is non-toxic, less time and fund consuming, efficient and a well-suited method for routine DNA isolation from Gram negative bacteria. [ABSTRACT FROM AUTHOR]

Purpose: Corn silk is a fiber-rich byproduct obtained during corn processing. The present investigation deals with the purpose of GMO detection and quantification and comparison of DNA extraction in corn silk which is considered generally a waste product. Method: A comprehensive analysis of extracted DNA quality, extraction cost, time, and PCR amplification was performed. Samples were homogenized using three different methods i.e. tissue lyser (CSK-TL) liquid nitrogen (CSK-LN), and grinder (CSK-RG). The homogenized samples from different methods were subjected to scanning electron microscopy (SEM) to access the effectiveness of homogenization methods. Afterward, DNA extraction was carried out using three different Kits QIAGEN DNeasyMericon Food Kit, eurofins|GeneScan GENESpin DNA isolation Kit, and PENICON EZ-10 Spin Column Genomic DNA Miniprep Kit. Furthermore, the fluorometric assay was done using Qubit fluorometer 3.0 for quantification of the extracted DNA. The quality of the DNA was accessed on 1.5% agarose gel. Results: The scanning electron microscopy (SEM) technique displays observable differences among the three homogenization methods. As compared to the other two methods tissue lyzer (CSK-TL) shows the finest particle image. Whereas, the images of liquid nitrogen (CSK-LN), as well as a grinder (CSK-RG) treated sample, show granulated particles. DNA extraction of homogenized samples was carried out using three different kits named QIAGEN DNeasyMericon Food Kit (Kit 1), eurofins|GeneScan GENESpin DNA isolation Kit (Kit 2), and PENICON EZ-10 Spin Column Genomic DNA Miniprep Kit (Kit 3). Kit 1 showed a yield of 122.4 ng/µl, whereas Kit 2 and Kit 3 gave 99.6 and 40.1 ng/µl respectively. By using gel electrophoresis technique the band size of corn silk genomic DNA was above 1000 bp. Conclusion: DNA extracted using waste material (corn silk) can be used for GMO analysis through the Real-Time PCR technique. Among different grinding techniques compared, the Tissue lyzed sample (CSK-TL) displayed the best homogenization and the highest quantity of DNA using the QIAGEN DNeasyMericon Food Kit. This method demonstrates the utility of the waste products for tracing the GMO status. [ABSTRACT FROM AUTHOR]

A new UGT1A1*28 detection method combining PCR and high-resolution agarose gel electrophoresis was developed. The viability of this method was demonstrated on 15 healthy adult volunteers. Subjects included 13 wild type homozygotes (86.7 %), 2 heterozygotes (13.3 %), and no mutant type homozygotes (0 %). The new UGT1A1*28 detection method results were fully consistent with DNA sequencing. PCR and agarose gel electrophoresis are common techniques with high-resolution agarose gels available commercially. These results support the clinical viability of this method potentially reducing UGT1A1*28 diagnosis complexity and cost.

Two simple and inexpensive in-house qualitative human immunodeficiency virus type 1 nucleotide amplification tests (HIV-1 NATs) were established as adjunct confirmatory HIV test for HIV antigen (Ag)-positive specimens identified from HIV screening test and for patients with indeterminate or negative HIV antibody (Ab) confirmatory test results. The limit of detection was

As threats to amphibian health increase, there is a growing need for diagnostic tools to assess and monitor their health status. Plasma protein electrophoresis has proven to be useful in other nonmammalian species. It enables quantification of protein fractions in plasma that may be altered in various disease processes, and is therefore useful in narrowing down differential diagnoses and detecting inflammation, in combination with other modalities such as biochemical and hematologic testing. The amphibian electrophoretogram must be defined before baseline reference intervals are obtained across species. Agarose gel electrophoresis was performed on plasma samples collected from presumed clinically normal individuals of one anuran and six urodelans: Osteopilus septentrionalis (n = 2), Gyrinophilus porphyriticus (n = 1), Notophthalmus viridescens (n = 1), Eurycea guttolineata (n = 2), Amphiuma tridactylum (n = 2), Cryptobranchus alleganiensis (n = 5), and Siren lacertina (n = 6). The electrophoretograms varied in number of fractions between each species; however, the number of fractions was consistent within a species. An albumin migrating fraction was consistently observed in all species. A prealbumin migrating fraction was identified in species that primarily use organs other than skin for respiration. This study provides preliminary examples of a normal plasma protein electrophoretogram for seven amphibian species. Further studies quantifying reference intervals and identification of protein fractions will help establish protein electrophoresis as a useful tool in amphibian health investigations. [ABSTRACT FROM AUTHOR]

Amyloid-associated neurodegenerative diseases, including Alzheimer’s disease (AD), are characterized by the in-brain accumulation of β-sheet structured protein aggregates called amyloids. However, neither a disease model nor therapy is established. We review past data and present new, preliminary data and opinions to help solve this problem. The following is the data-derived model/hypothesis. (1) Amyloid-forming proteins have innate immunity functions implemented by conversion to another sheet conformation, α-sheet. (2) In health, α-sheet structured, amyloid-forming proteins inactivate microbes by co-assembly with microbe α-sheets. Amyloid-forming proteins then undergo α-to-β-sheet conversion. (3) In disease, α-sheet-structured, amyloid-forming proteins over-accumulate and are neuron-toxic. This hypothesis includes formation by virus capsid subunits of α-sheets. In support, we find that 5–10 mM methylene blue (MB) at 54 °C has a hyper-expanding, thinning effect on the phage T4 capsid, as seen by negative stain- and cryo-electron microscopy after initial detection by native gel electrophoresis (AGE). Given the reported mild anti-AD effect of MB, we propose the following corollary hypothesis. (1) Anti-AD MB activity is, at least in part, caused by MB-binding to amyloid α-sheet and (2) MB induces the transition to α-sheet of T4 capsid subunits. We propose using AGE of drug incubated T4 to test for improved anti-AD activity.

Simple Summary: Serum protein electrophoresis (SPE) is a widely used method to determine the concentration of a serum protein fraction in human and veterinary medicine. Serum protein electrophoresis allows the separation of serum proteins into several categories based on their ability to migrate in an electrical field on different supports. Typical migration patterns could be identified in the course of different disorders such as inflammation or neoplasia. However, to be clinically useful, the interpretation of SPE data had to be performed in light of an appropriate method—and species-specific reference intervals (RIs) and for all species, including marine mammals, attention should also be given to the different living environments. In this work, we established the RIs for serum protein fractions evaluated using agarose gel electrophoresis in bottlenose dolphins (Tursiops truncatus) under human care. These data are a new tool for health assessment in dolphins, thus, they may be useful for clinicians to better interpret laboratory data. Serum protein electrophoresis (SPE) is the most used and reliable method to determine the percentage of serum protein subfractions. The interpretation of the kinetics of total proteins and albumin and globulin fractions is receiving increased attention in wild animals, as well as in domestic animals, due to the possibility of identifying typical pathologic patterns. However, the interpretation of these data had to be performed in light of an appropriate method—and species- specific reference intervals (RIs). In marine mammals, as well as other non-domestic species, specific attention should also be given to the different environment (free ranging vs. human managed) and the associated different exposure to environmental stimuli. The aim of this report was to establish RIs for the serum protein fractions evaluated using agarose gel electrophoresis (AGE) in bottlenose dolphins under human care. Peripheral blood samples were collected from 40 bottlenose dolphins during standard veterinary procedures to evaluate their health status. Total protein concentration was determined using the biuret method while AGE was performed using an automated system. A pooled dolphin's serum sample was used to determine the intra-assay and inter-assay imprecision of AGE. The RIs were calculated using an Excel spreadsheet with the Reference Value Advisor set of macroinstructions. The intra and inter-assay imprecisions were 1.2% and 2.5%, respectively, for albumin; 2.9% and 5.7%, respectively, for α-globulins; 3.8% and 4.0%, respectively, for β-globulins; and 3.4% and 4.8%, respectively, for γ-globulins. The total protein, albumin, α-globulin, β-globulin, and γ-globulin concentrations were 65.5 ± 5.4 g/L, 45.5 ± 4.9 g/L, 8.0 ± 1.0 g/L, 5.0 ± 2.0 g/L, and 7.0 ± 2.0 g/L, respectively. We established the RIs for the total protein and serum protein fractions using AGE in bottlenose dolphins under human care. [ABSTRACT FROM AUTHOR]

We demonstrate that gDNA can be conveniently and efficiently isolated and purified using standard agarose gel electrophoresis, band excision and gel purification. This method yields a substantial amount at microgram levels of gDNA per gel cleanup with high purity. An RNase A treatment step can be omitted. The quality of gDNA is suitable for next-generation sequencing, resulting in >10 Mb reads and high-quality read data (Phred score >28 up to 100 of 150 base reads). Furthermore, the gDNA can be kept intact in a gel slice for several days. This method has been tested for dictyostelids, bacteria and plants.

Copyright of China Tropical Medicine is the property of China Tropical Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Agarose gel electrophoresis is a relatively easy to use method, commonly applied to evaluate PCR reaction success. Intercalating agents or dyes are used to visualize the amplified fragments. However, it is uncertain to what extent the brightness of bands is informative about the concentration of the amplicons. To more closely examine the suitability of agarose gel electrophoresis to assess PCR product yield, we quantified the brightness of bands on a gel and compared these data with the results from spectrophotometry, fluorometry and qPCR. Evaluation of the results suggests that assessment of the relative quantity of amplicons by band brightness is precise enough even for post-PCR analysis steps requiring PCR product concentrations within a certain range to function properly.

Simple Summary: Rice, the staple food for more than half of humanity, is grown predominantly in Asia, the world's most populous continent with the fastest-growing economy. The present-day rice industry must not only meet increasing demand but also changing consumer demands, with a strong emphasis placed on producing high-quality rice. While the rapid development of advanced genotyping methods can be useful for modern rice breeding programs, some methods (such as capillary electrophoresis or sequencing) can be costly to apply in laboratories with limited resources. To address this issue, we developed six novel multiplex polymerase chain reaction (PCR) assays that employ a standard agarose-based gel electrophoresis system to simultaneously detect at least two major grain quality (amylose content and fragrance) and biotic stress (blast, sheath blight, and bacterial leaf blight) genes in rice. One of these assays, which can detect all three targeted biotic stresses, was found to be useful in screening Malaysian weedy rice that may contain novel sources of disease resistance. The universal protocol described in this study can be used in routine molecular laboratories to aid rice breeding initiatives in Malaysia and other resource-constrained countries. While previous research has demonstrated that multiplex polymerase chain reaction (PCR) can be a cost-effective approach to detect various genes in crops, the availability of multiplex assays to simultaneously screen both grain quality and biotic stress resistance traits in rice (Oryza sativa) is limited. In this work, we report six novel multiplex assays that use a universal protocol to detect major rice grain quality (amylose content and fragrance) and biotic stress (blast, sheath blight, and bacterial leaf blight) traits with amplified products consisting of up to four primer pairs that can be analyzed using a standard agarose-based gel electrophoresis system. Recent studies have suggested that weedy rice has novel sources of disease resistance. However, an intensive screening of weedy biotypes has not been reported in Malaysia. Accordingly, we employed one of the developed multiplex assays to screen reported genes or quantitative trait loci (QTLs) associated with blast, sheath blight, and bacterial leaf blight diseases in 100 weedy rice biotypes collected from five local fields, with phenotyping performed to validate the genotyping results. In conclusion, our universal multiplex protocol is effective for the large-scale genotyping of rice genetic resources, and it can be employed in routine molecular laboratories with limited resources. [ABSTRACT FROM AUTHOR]

Various dyes are used to visualize DNA bands in agarose gel electrophoresis (AGE) by the methods of pre- or post-staining. The DNA dye user's guides generally state that the binding of the dye to DNA will affect DNA mobility in electrophoresis, thus recommending post-staining for accurate measurement of DNA size. However, many AGE performers prefer pre-staining procedures for reasons such as convenience, real-time observation of DNA bands, and/or the use of a minimal amount of dye. The detrimental effect of the dye on DNA mobility and the associated risk for inaccurate measurement of DNA size are often overlooked by AGE performers. Here we quantitatively determine the impact on DNA migration imposed by frequently used dyes, including GelRed, ethidium bromide (EB), and Gold View. It was observed that pre-staining with GelRed and EB significantly slowed down DNA migration to cause as much as 39.1% overestimation on the size of sample DNA, whereas Gold View had little effect. The slowdown of DNA migration increased with dye concentration until it plateaued when the dye concentration reached a saturated level. Thus, to take advantage of pre-staining, saturated levels of DNA dyes should always be applied for both DNA samples and DNA markers to ensure a fair comparison of DNA sizes. In addition, GelRed and EB display much higher sensitivity than Gold View in the detection of DNA bands in post-staining. The saturated concentrations, cost considerations, and other useful features of these frequently used dyes are summarized for the information of AGE performers., (© 2024 Wiley‐VCH GmbH.)

DNA sequencing is crucial in molecular biology techniques to identify Staphylococcus aureus isolates' genotype to advance diagnoses, prevent, and control infections. DNA isolation can be influenced by several factors which can affect the quality of the output. The experiment was conducted to assess the quality and quantity of genomic DNA isolated from S. aureus. The gDNA's purity and concentration were determined using the spectrophotometric assay, and the quality was analyzed using PCR and agarose gel electrophoresis. The purity of DNA and RNA was determined using the absorbance ratio at A260 and A280, and the percent coefficient of variation was used for the three replicates. The result showed that the extracted gDNA from S. aureus was obtained in large quantities, with good quality, R1=93.67 µg/ml, R2=59.85 µg/ml, and R3=67.50 µg/ml. The concentration was within a 1.8 – 2.0 absorbance ratio and was considered high quantity output. The extracted and amplified pure gDNA fragment from the sample can now be used for other molecular techniques.

Serum protein electrophoresis (SPE) is the most used and reliable method to determine the percentage of serum protein subfractions. The interpretation of the kinetics of total proteins and albumin and globulin fractions is receiving increased attention in wild animals, as well as in domestic animals, due to the possibility of identifying typical pathologic patterns. However, the interpretation of these data had to be performed in light of an appropriate method—and species- specific reference intervals (RIs). In marine mammals, as well as other non-domestic species, specific attention should also be given to the different environment (free ranging vs. human managed) and the associated different exposure to environmental stimuli. The aim of this report was to establish RIs for the serum protein fractions evaluated using agarose gel electrophoresis (AGE) in bottlenose dolphins under human care. Peripheral blood samples were collected from 40 bottlenose dolphins during standard veterinary procedures to evaluate their health status. Total protein concentration was determined using the biuret method while AGE was performed using an automated system. A pooled dolphin’s serum sample was used to determine the intra-assay and inter-assay imprecision of AGE. The RIs were calculated using an Excel spreadsheet with the Reference Value Advisor set of macroinstructions. The intra and inter-assay imprecisions were 1.2% and 2.5%, respectively, for albumin; 2.9% and 5.7%, respectively, for α-globulins; 3.8% and 4.0%, respectively, for β-globulins; and 3.4% and 4.8%, respectively, for γ-globulins. The total protein, albumin, α-globulin, β-globulin, and γ-globulin concentrations were 65.5 ± 5.4 g/L, 45.5 ± 4.9 g/L, 8.0 ± 1.0 g/L, 5.0 ± 2.0 g/L, and 7.0 ± 2.0 g/L, respectively. We established the RIs for the total protein and serum protein fractions using AGE in bottlenose dolphins under human care.

We tested the hypothesis that age, breed, and sex are related to hematology, biochemistry, acute phase proteins (APPs), seroreactivity and level of parasitemia in dogs with an acute phase response (APR) due to Babesia canis infection. The study enrolled 61 privately owned dogs that naturally acquired B. canis infection. Groups were formed according to the age: young dogs less than one year, and adult dogs more than one year old. Moreover, the group of males was compared to females and purebred to mixed breed dogs. Seroreactivity was tested with immunofluorescence antibody test, level of parasitemia with real-time polymerase chain reaction (real-time PCR), hematology, and biochemistry with automatic analyzers, serum amyloid A with enzyme-linked immunosorbent assay, fibrinogen with heat precipitation and ceruloplasmin and paraoxonase-1 with manual spectrophotometric methods. For protein separation agarose gel electrophoresis was used. The main changes in the whole population of B. canis -infected dogs were fever, pancytopenia, and change in APPs level. One-third of young, and 96% of adult dogs were seropositive (P < 0.001). The level of parasitemia was higher in the young dogs (P < 0.001). Erythroid lineage parameters (P < 0.01), and leukocytes (P < 0.05) were lower in the young, when compared to the adult dogs. Young dogs had lower total globulins (P < 0.001), β- and γ-globulins (P < 0.001), and higher α-globulins (P = 0.022) than adult dogs. Young dogs had higher concentrations of phosphate (P = 0.003) and cholesterol (P < 0.001) and lower amylase (P = 0.014) and lipase activity (P = 0.020) than adult ones. Male dogs had lower neutrophil count than females (P = 0.035), and purebred dogs had more band neutrophils than mixed breed dogs (P = 0.004). In conclusion, dogs with natural Babesia canis infection at a young age have more severe anemia and APR including leukopenia than adults. Male and purebred dogs might also have more severe APR than females and mix-breeds, as they have more pronounced changes related to the myeloid lineage. • Young dogs have more severe inflammation due to B. canis infection than adult dogs. • More male dogs become neutropenic due to B. canis infection than female dogs. • Purebreds show a more severe left shift due to B. canis infection than mixed-breeds. [ABSTRACT FROM AUTHOR]

Human tissue samples soaked in formaldehyde solution are rare resources in many cases, and the extraction of their DNA is a challenging issue. In this paper, genomic DNA was extracted from four formalin soaked human embryo skin and muscle samples by using an improved method for extracting genomic DNA from samples soaked in formaldehyde solution and a generic genomic DNA extraction kit. The purity and concentration of extracted DNA were determined by UV spectrophotometry, and the DNA solution was subjected to agarose gel electrophoresis and imaging. The results showed that the concentration and purity of genomic DNA obtained by the improved extraction method were better than those obtained by the general method. PCR amplification was carried out with the extracted genomic DNA template. Electrophoresis results showed that the improved extraction method had better PCR amplification effect. Our study provides methodological guidance and a reference for the direction of improvement for the extraction of genomic DNA from precious samples soaked in formaldehyde solution [ABSTRACT FROM AUTHOR]

The information about the clinical features of Leishmania infantum infection in cats is scarce. In this study, we evaluated the serum protein electrophoresis of samples from 19 infected but apparently healthy cats. To detect L. infantum infection, two serological tests, i.e. western blot (WB) and enzyme-linked immunosorbent assay (ELISA) as well as quantitative polymerase chain reaction (qPCR) on the blood samples were performed. Eventual infection by several selected bacterial and viral pathogens was also tested. All but one of the cats were found positive with WB. The WB-negative cat was positive by ELISA only. From the 18 WB-positive cats, only three were positive also by ELISA and eight with qPCR, including the only animal which was positive in all the three tests. No concomitant infections were detected in any of the cats. The main alteration of the proteinogram was characterised by an increase of the α-2 fraction. In the five cats with hypergammaglobulinaemia, the pattern detected was polyclonal. None of the cats were seropositive to any other pathogens tested. The presence of polyclonal gammopathy and elevation of the α-2 fraction could suggest the presence of active infection. In contrast, the only detection of an increase of the α-2 fraction alone with the presence of positive serological result could be associated by immune response activation against L. infantum. [ABSTRACT FROM AUTHOR]

EPH has been demonstrated to be a useful tool in companion animals while providing an opportunity to characterize globulinemias, including paraproteinemia. In EPH of non‐traditional species, these same applications are important, but the primary use is to gauge the acute‐phase and humoral immune responses. This includes the valid quantitation of albumin as well as the examination of fractions reflective of increases in acute‐phase reactants and immunoglobulins. Agarose gel EPH and, more recently, capillary zone EPH have been applied to samples from these species. Performing these analyses provides special challenges in the placement of fraction delimits, generation of RIs, and interpretation of results. Recommended as part of routine bloodwork, EPH can also provide key results that are helpful in clinical and field‐based health assessments as well as in prognostication. [ABSTRACT FROM AUTHOR]

Background: Densitometric quantification of myeloma paraproteins (M‐proteins) is used to monitor secretory myeloma related disorders in humans and dogs. The previous work in dogs used agarose gel electrophoresis (AGE) but did not establish if other methods of serum protein electrophoresis, such as capillary zone electrophoresis (CZE), were comparable. Objectives: We aimed to determine if the densitometric quantification of M‐proteins using CZE would yield results comparable to AGE methods. Methods: Fifty‐one serum samples from 22 dogs and 18 cats with confirmed monoclonal gammopathies and previously performed AGE were evaluated using CZE on a Sebia Minicap system. Samples were run in duplicate, and their M‐proteins were densitometrically measured using the corrected perpendicular drop method previously described. Human‐based quality control samples were used to determine the inter‐run coefficient of variation (CV). Patient samples were used to calculate the intra‐run CV. Method comparison was performed using simple linear regression, Passing‐Bablok regression, and Bland‐Altman analyses, and Medx evaluations. Results: Inter‐run and intra‐run CVs for CZE were 3.71%‐7.65% and 2.89%‐4.74%, respectively. Simple linear regression demonstrated an excellent correlation (r > 0.98). Passing‐Bablok regression was compatible with the presence of proportional bias in the entire population, and Bland‐Altman plots revealed a proportional bias in the feline cases. The Medx evaluation suggested that the two methods did not perform similarly in clinical samples with poor performance at a decision limit of 0.5 gm/dL. Conclusions: Capillary zone electrophoresis is an acceptable method for M‐protein densitometric quantification in canine and feline sera but cannot be used interchangeably with AGE‐based evaluations. [ABSTRACT FROM AUTHOR]

In this paper, a copper complex [CuL2](PF6)3·H2O(complex 1) was synthesized using 1-methyl-4′-(p-tolyl)-2,2′,6′,2″-terpyridine hexafluorophosphate (L) as the ligand. The structure of the synthesized complex was characterized by X-ray single crystal diffraction technique, thermogravimetric analysis and X-ray photoelectron spectroscopy (XPS). The results of single crystal diffraction analysis show that the complex belongs to the monoclinic crystal system, the P1/c space group, and the unit cell parameters were a=1.273 15(10) nm, b=1.902 01(13) nm, c=2.109 55(17) nm, β=101.269(2)° and V=5.009 9(7) nm3. The XPS analysis result shows that the copper ions in complex 1 exists in the form of +1 valence. Thermogravimetric analysis shows that the complex has good thermal stability. DNA photocleavage studies monitored by agarose gel electrophoresis shows that the complex cleaved DNA efficiently, which indicates that the complex has obvious potential biological activity. [ABSTRACT FROM AUTHOR]

Objective: To breed and identify inducible forebrain-specific cholecystokinin receptor-2 (CCKR-2) transgenic (tTA/tetO-CCKR-2 tg, abbreviated as dtg) mice, in order to serve the dtg mice as models for further studying the molecular mechanism and pathogenesis of CCKR-2 in fear behavior, anxiety disorder, posttraumatic stress disorder morbidity and other related diseases. Methods: (1) The dtg mice were obtained by crossing α-CaMKII/tTA single transgenic mice with tetO/CCKR-2 single transgenic mice, and the offspring was obtained as distant relatives as possible. The genomic DNA was extracted from the tails tissue of the offspring mice. PCR and agarose gel electrophoresis were used to identify the PCR products. (2) In situ hybridization verified the specific expression of CCKR-2 double transgenic mice in the forebrain, and that dtg mice were selected as subculture and experimental mice. Results: (1) The clear bands of tTA (350 bp) and CCKR-2 (550 bp) were shown in the PCR gel electrophoresis pattern, while no band was shown in the wild type, which indicated that the agarose gel electrophoresis results were consistent with the expected target size of gene fragments in the dtg mice model. (2) In situ hybridization results showed that CCKR-2 was strongly expressed in the forebrain of dtg mice, but not in the wild type mice. The results showed that dtg double transgenic mice were successfully bred and identified in the laboratory, and more dtg mice were bred. Conclusions: The dtg double transgenic mice can be obtained successfully through correct breeding and gene identification, which provides references for the application of the model and lays a foundation for the follow-up studies in the laboratory. [ABSTRACT FROM AUTHOR]

The present research describes the synthesis, characterization, in vitro biological and docking evaluation of N,N'-(ethane-1,2-diyl)bis(benzamides) (3a-3j). Consequently, in in vitro hRBCs hemolysis assay, only the bis-amide (3d) induced 52.4% hemolysis at higher concentration (1000 μg/mL) that decreased drastically with concentration (250 μg/mL) to 27.9% (CC50 = 400.41). Similarly, the tested bis-amide (3j) was found to be the least toxic with 7.8% hemolysis at higher concentration (1000 μg/mL) that gradually decreases to 6.1% (CC50 = 19,347.83) at lower concentration (250 μg/mL). Accordingly, the tested bis-amides were found to be highly biocompatible against hRBCs at higher concentrations with much higher CC50 values (> 1000 μg/mL). The biocompatible bis-amides (3a-3j) were subjected to in vitro DNA ladder assay to analyze their apoptotic potential. The results obtained suggest the tested bis-amides (3a-3j) are highly degradative toward DNA causing the appearance of more than one bands or complete degradation of DNA except (3a), (3c), (3i) and (3 g). Moreover, the synthesized bis-amides (3a-3j) were tested in in vitro antileishmanial assay to unveil their leishmaniacidal potential. The results obtained clearly indicated that some of the tested bis-amides displayed good dose dependent response. The tested bis-amides were highly active at higher concentration (1000 μg/mL) against the leishmanial promastigotes and their % inhibitory potential decreased drastically with concentration (250 μg/mL). Consequently, at higher concentration (1000 μg/mL), the bis-amide (3f) caused 85% inhibition and was ranked as the most effective leishmaniacidal bis-amides followed by the bis-amide (3 g) with 73.54% inhibition of leishmanial promastigotes. However, in terms of their IC50 values, the best leishmaniacidal potential was displayed by the bis-amide (3f) followed by (3b), (3j) and (3 g) with IC50 values increasing in the order of 633.16, 680.22, 680.22 and 712.93 μg/mL, respectively. Molecular docking studies revealed that bis-amides having electron-donating groups showed good binding potential against antileishmanial target. [ABSTRACT FROM AUTHOR]

While previous research has demonstrated that multiplex polymerase chain reaction (PCR) can be a cost-effective approach to detect various genes in crops, the availability of multiplex assays to simultaneously screen both grain quality and biotic stress resistance traits in rice (Oryza sativa) is limited. In this work, we report six novel multiplex assays that use a universal protocol to detect major rice grain quality (amylose content and fragrance) and biotic stress (blast, sheath blight, and bacterial leaf blight) traits with amplified products consisting of up to four primer pairs that can be analyzed using a standard agarose-based gel electrophoresis system. Recent studies have suggested that weedy rice has novel sources of disease resistance. However, an intensive screening of weedy biotypes has not been reported in Malaysia. Accordingly, we employed one of the developed multiplex assays to screen reported genes or quantitative trait loci (QTLs) associated with blast, sheath blight, and bacterial leaf blight diseases in 100 weedy rice biotypes collected from five local fields, with phenotyping performed to validate the genotyping results. In conclusion, our universal multiplex protocol is effective for the large-scale genotyping of rice genetic resources, and it can be employed in routine molecular laboratories with limited resources.

Abstract Von Willebrand factor (VWF) is a 500‐ to 15 000‐kDa multimeric protein circulating in the blood. When VWF has a higher molecular weight, its hemostatic activity is greater. The size distribution of VWF multimers is usually analyzed by SDS‐agarose gel electrophoresis followed by immunoblotting. We found that the most commonly used anti‐VWF antibody cross‐reacted with fibronectin in VWF multimer analysis. In addition, since the apparent molecular weights of VWF and fibronectin are almost identical, these molecules were difficult to distinguish by SDS‐polyacrylamide gel electrophoresis followed by immunoblotting. Cross‐reactivity between the anti‐VWF antibody and fibronectin was inhibited by pretreating the antibody with fibronectin‐coated plates. To obtain accurate data using anti‐VWF antibodies, it is necessary to be aware of the possibility of cross‐reactivity with fibronectin.

This work assessed the catabolic versatility of functional genes in hydrocarbon-utilizing bacteria obtained from the rhizosphere of plants harvested in aged polluted soil sites in Ogoni and their attenuation efficacy in a bioremediation study. Rhizosphere soil was enumerated for its hydrocarbon-utilizing bacteria. The bacteria were in-vitro screened and selected through the quantification of their total protein and specific intermediate pathway enzyme (catechol 2,3-dioxygenase) activity in the metabolism of hydrocarbon. Thereafter, agarose gel electrophoresis technique was deployed to profile the genome of the selected strains for catechol 2,3-dioxygenase (C23O), 1,2-alkane monooxygenase (alkB), and naphthalene dioxygenase (nahR). Four rhizobacterial isolates namely Pseudomonas fluorescens (A3), Achromobacter agilis (A4), Bacillus thuringiensis (D2), and Staphylococcus lentus (L1) were selected based on the presence of C23O, alkB, and nahR genes. The gel electrophoresis results showed an approximate molecular weight of 200 bp for alkB, 300 bp for C23O, and 400 bp for nahR. The gas chromatogram for residual total petroleum hydrocarbon (TPH) revealed mineralization of fractions C8–C17, phytane, C18–C30. TPH for in-vitro bioremediation of crude oil-polluted soil was observed to have an optimal reduction/loss of 97% within the 56th day of the investigation. This study has further revealed that the microbiome of plants pre-exposed to crude oil pollution could serve as a reservoir for mining group of bacterial with broad catabolic potentials for eco-recovery and waste treatment purposes. [ABSTRACT FROM AUTHOR]

The present study is focused on application of a natural compound, 3, 5-dihydroxy 4′, 7-dimethoxyflavone (DHDM) from a medicinal plant Alpinia nigra for nucleic acid detection and differential cell staining. DHDM was found to interact with nucleic acid and forms complex, which was investigated for various applications. It was successfully utilized to visualize plasmid, genomic, and ds-linear DNA in agarose gel electrophoresis without affecting the DNA mobility in the gel. Fluorescence of DHDM increased several fold upon binding to dsDNA. Photostability of the compound was assessed and showed photobleaching effect that decreased gradually over time. Application of the compound was further extended to differential cell staining. When observed in fluorescence microscope, DHDM stained the dead cells and differentiated them from live cells in the case of bacterial, yeast, and mammalian cells. Higher concentration of the compound was found to be less cytotoxic to cancerous cells. Nucleic acid staining dyes like Ethidium bromide (EtBr), Propidium iodide (PI), etc. are carcinogens and environmental pollutants and therefore DHDM a natural compound, is a major benefit and thus can serve as an alternative to the current dyes. • The 3, 5-dihydroxy 4′, 7-dimethoxyflavone (DHDM) displayed the binding potential of nucleic acid (DNA). • Fluorescence study revealed the binding confirmation with DNA. • DHDM has played a vital role in live-dead cell differentiation. • Fluorescence microscopy was used to quantify staining to live-dead cell differentiation. • This study might be helpful for the development of new dye for DNA and live-dead cell differentiation. [ABSTRACT FROM AUTHOR]

The article presents a case study of a 53-year-old man with diagnosis of type 1 diabetes mellitus and various diabetes complications such as nephropathy, mild retinopathy, and moderate sensory-motor neuropathy. Topics include blood revealing renal insufficiency with secondary hyperparathyroidism and increased hemoglobin; and quantifying the protein fractions based on protein-specific dye binding.

Serum protein electrophoresis is commonly used in veterinary medicine to obtain a broad picture of acute phase and humoral immune responses. Agarose gel electrophoresis (AGE) has been the most prevalent platform in laboratory settings; however, in human medicine, this method is becoming increasingly supplanted by capillary zone electrophoresis (CZE). CZE has been demon-strated to have superior accuracy in fraction quantitation as well as consistently resolved protein fractions that are generally not observed by AGE. The objectives of the current study were to compare these two methods using serum from bottle-nose dolphins (Tursiops truncatus) and to generate preliminary reference intervals for this species. The methods were compared using Spearman's correlation, Passing-Bablok regression, and Bland Altman analysis. A varying but significant correlation was observed between the methods for all protein fractions and the albumin/globulin (A/G) ratio except for the beta globulin fraction. Comparable or better imprecision in fraction quantitation was purobserved for CZE. Constant and proportional error was detected for both alpha globulin fractions. Of note, CZE allowed for an easier definition of the beta globulin fraction into two fractions (beta 1 and beta 2 globulins), and other subfractions were also visible. In total, these statistical and visible differences indicate that the two methods are not equivalent which necessitates the generation of method-specific reference intervals. Additionally, as it becomes more widely available, CZE should be considered as the preferred method for electrophoresis in this species. [ABSTRACT FROM AUTHOR]

The electrochemical oxidation of 3‐nitro‐tyrosine (3‐NO2‐Tyr) was studied in aqueous media at metallic electrodes (platinum and gold), using voltammetric techniques. The interaction between 3‐NO2‐Tyr and double helix DNA (dsDNA) in a physiological medium was also investigated. Electro‐oxidation of 3‐NO2‐Tyr occurs in one single irreversible pH‐dependent step with the transfer of one electron and one proton from the phenolic group to the formation of radicals, which preferably dimerize, fouling the electrode surfaces. The differential pulse voltammetry and gel electrophoresis results clearly demonstrated a strong interaction of 3‐NO2‐Tyr with the dsDNA for the formation of a stable 3‐NO2‐Tyr‐dsDNA complex. [ABSTRACT FROM AUTHOR]

A new UGT1A1*28 detection method combining PCR and high-resolution agarose gel electrophoresis was developed. The viability of this method was demonstrated on 15 healthy adult volunteers. Subjects included 13 wild type homozygotes (86.7 %), 2 heterozygotes (13.3 %), and no mutant type homozygotes (0 %). The new UGT1A1*28 detection method results were fully consistent with DNA sequencing. PCR and agarose gel electrophoresis are common techniques with high-resolution agarose gels available commercially. These results support the clinical viability of this method potentially reducing UGT1A1*28 diagnosis complexity and cost., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Abstract This study aimed to develop a simple and rapid method to detect KRAS gene mutations for conventional clinical applications under laboratory conditions. The genotype of mutation sites was determined based on the occurrence of target bands in the corresponding lanes of the reaction tubes through polymerization-conjunction of the probes, probe purification and amplification, and agarose gel electrophoresis. Circulating DNA samples were obtained from the plasma of 72 patients with lung cancer, which were identified based on six mutation sites (G12S, G12R, G12C, G12D, G12A, and G12V) of codon 12 of the KRAS gene. The detection results were compared with direct sequencing data. The proposed detection method is characterized by simple operation, high specificity, and high sensitivity (2%). This method can detect the mutations of three samples at G12S, G12R, and G12A. In the direct sequencing spectra of these samples, the genotype could not be determined due to the lack of evident sequencing peaks that correspond to the basic group of mutations. In conclusion, a simple and rapid method was established based on probe polymerization-conjunction-agarose gel electrophoresis for detecting KRAS gene mutations. This method can be applied to the conventional mutation detection of inhomogeneous samples.

Introduction. Analysis of canine lipoprotein fractions after agarose gel electrophoresis (agEF) separation could be an important diagnostic tool in primary and secondary dyslipidemia diagnosis. The aim of this study was to measure concentrations of triglycerides and cholesterol and to analyze lipoprotein fractions in dogs after basal (12 hours) and short-term (24 and 36 hours) fasting, i.e., frequent conditions in clinical practice. Materials and Methods. Blood samples were collected from six lean dogs of both sexes and different breeds, after 12, 24 and 36 hours of fasting. Concentrations of glucose, triglycerides and cholesterol were determined on an automated wet biochemistry analyzer, lipoprotein fractions were separated by agEF and leukocyte numbers were assessed on an automated hematology analyzer. Results and Conclusions. Results showed there was no significant change in glucose, triglyceride and cholesterol concentrations nor in leukocyte numbers during dog fasting. Moreover, there was no change in α1- and α2-fractions, but there was a significant decrease in preβ- and β-fraction of lipoproteins. It is know that high density lipoproteins (HDL) have α-mobility and very low density (VLDL) and low density lipoproteins (LDL) have preβ- and β-mobility. Thus, it is possible that reverse cholesterol transport maintained by HDL is not affected during short-term fasting. On the contrary, synthesis of VLDL and formation of LDL are probably decreased because endogenous synthesis of triglycerides is decreased or their clearance is increased.

Background: Many kinds of sulfated glycosaminoglycans (GAGs) play an essential role in both physiological and pathological conditions. Most of them are obtained from animal sources, and used as nutraceuticals or therapeutic applications. Objectives: In this study, we aimed to screen for the presence of sulfated GAGs from 8 locally available plants, including ginger (Zingiber officinale), turmeric (Curcuma longa), star fruit (Averrhoa carambola), Namwa banana (Musa ABB ‘Kluai Namwa’), bitter gourd (Momordica charantia), purple-fruited pea eggplant (Solanum trilobatum), noni (Morinda citrifolia), and finger root (Boesenbergia rotunda). Materials and methods: All plants were extracted, and sulfated GAGs from the extracts were investigated by dimethylmethylene blue (DMMB) dye-binding assay, uronic acid assay, UV-Vis spectrophotometry, and agarose gel electrophoresis. Results: DMMB dye-binding and uronic acids assays revealed the presence of sulfated GAGs in all extracts with various degrees of sulfated GAGs levels. These results correlated to UV-Vis spectrophotometry that showed the maximum absorbance peaks from all extracts at 190-210 nm, which was similar to sulfated GAGs standard absorption. Interestingly, agarose gel electrophoresis suggested that sulfated GAGs in all extracts exhibited diverse patterns in alcian blue, toluidine blue and safranin O staining. Conclusion: Our results indicate that all plant extracts contain sulfated GAGs at certain levels, which could be a new approach for future study in bioprospecting.

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Agarose gel electrophoresis (AGE) has been widely implemented throughout veterinary medicine and for analysis of plasma proteins of avian and reptile species. Capillary zone electrophoresis (CZE) is becoming a standard method in human clinical pathology laboratories but has not widely been used for the analysis of animal samples. The objective of the present study was to compare protein fractions derived from AGE and CZE methods using plasma from the green turtle (Chelonia mydas). Plasma samples were analyzed by AGE and CZE per manufacturer guidelines. The methods were assessed by CV analysis, Spearman's correlation, Passing-Bablok regression, and Bland Altman plots. CZE consistently resolved more fractions than AGE with three fractions observed in the prealbumin migrating region versus one for AGE and two fractions in the c globulin region versus one for AGE. Compared with AGE, CZE showed a lower CV in intra-assay tests (1.0-4.9% vs 2.0-28.3%) and a lower or overlapping CV in interassay tests (1.0-10.6 vs 2.3-22.0). The prealbumin, a2 globulin, and b globulin fractions correlated the least between the methods (for all three fractions: rs ≤ 0.28, P . 0.21). Moderate, significant correlations between AGE and CZE methods were observed for albumin (rs ≤ 0.78, P, 0.0001) and c globulins (rs¼0.78, P, 0.0001). CZE has a higher precision and ease of use over AGE and offers the opportunity to resolve additional protein fractions. This will necessitate the development of new conventions in placement of fraction delimits, definition of species-specific reference intervals, and evaluation of clinical utility in abnormal turtles. [ABSTRACT FROM AUTHOR]

The DNA extraction process is the most crucial step for demonstrating various molecular techniques such as PCR, DNA barcoding, metagenomics etc. This paper describes a simple and efficient protocol of DNA extraction from insect's sample that included Honey bees, Carpenter bees and Flour beetle's tissues with slight modification in CTAB method. DNA concentration together with purity has been determined by spectrophotometer absorbance reading at O.D260nm and O.D260/280nm respectively. Additionally, lambda DNA marker (NEB3012S) has been employed to validate band intensity by agarose gel electrophoresis. The DNA band exposed less smearing and intense band in agarose gel. Band of DNA in HB and CB lane was greater than 95.4ng but less than 95.4ng in lane FB as compared to lambda DNA marker bands. Also, DNA extracted from the tissue of Honey bee (HB), Carpenter bee (CB) and Flour Beetle (FB) measured by spectrophotometer for analysing O.D at 260 nm was found to be 4.9, 5.1 and 3.7ug/ml and the purity by considering 260/280 O.D ratio was 2.33, 1.76, 1.92. Absorbance ratio of 2.33 in DNA of Honey bees (HB) sample indicated slight contamination of protein or carbohydrate. Therefore, an achievable approach has been made to extract high quality of DNA by CTAB method and applying spectrophotometer for Optical Density as well as marker lambda DNA by agarose gel electrophoresis for quantity and quality analysis. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Southern Agriculture is the property of Journal of Southern Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introduction: Considering the high endemicity of typhoid in developing country and the feasibility of phage therapy for this disease, the study was planned to determine the genomic structure of bacteriophages specific to ‘Vi’ positive Salmonella Typhi. Aim: To use different molecular tools to characterise Salmonella Typhi ‘Vi’ specific bacteriophages genotypically. Materials and Methods: Salmonella Typhi ‘Vi’ specific bacteriophages were isolated and propagated. Phage DNA was extracted and analysed by spectrophotometry, agarose gel electrophoresis and restriction enzyme digestion was done. Results: In the present study, two bacteriophages targeted against ‘Vi’ positive Salmonella Typhi were isolated. DNA of the phages was extracted for molecular characterisation. On restriction analysis, both the phages were found to be of similar pattern, suggesting that they were genetically similar, although they had different plaque morphologies. Conclusion: The present study may be helpful in genomic characterisation of Salmonella Typhi ‘Vi’ specific bacteriophages for their future use in therapeutics.

Two simple and inexpensive in-house qualitative human immunodeficiency virus type 1 nucleotide amplification tests (HIV-1 NATs) were established as adjunct confirmatory HIV test for HIV antigen (Ag)-positive specimens identified from HIV screening test and for patients with indeterminate or negative HIV antibody (Ab) confirmatory test results. The limit of detection was <1000 copies/mL, which is lower than that of the HIV Ag/Ab combination assay. One test using QL1 detected all 11 HIV-1 subtypes/circulating recombinant forms/group samples with almost equal analytical sensitivity, and the other test, using QL2, also detected all, except for two group O samples. In the examination of 28 HIV-1 Ag-positive samples using Determine HIV Early Detect, 27 samples were reactive and one HIV-1 Ag-pseudo-positive sample was non-reactive using both methods. These in-house qualitative HIV-1 NATs are useful for confirming HIV-1 Ag-positive cases and excluding HIV-1 Ag false-positive cases in areas with low HIV prevalence and small- and medium-sized diagnostic laboratories., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Protein electrophoresis and immunotyping can be a useful adjunct to the standard biochemical techniques for characterizing serum and urine proteins. This paper reviews currently available and commonly used methods for diagnostic protein electrophoresis, including both agarose gel and capillary zone electrophoretic techniques and total protein assessments. Immunofixation and immunosubtraction methods for identification of immunoglobulin location and class are also presented. Practical application of quality assurance and quality control strategies in compliance with American Society of Veterinary Clinical Pathology (ASVCP) best practices are discussed. Commonly encountered serum and urine electrophoretic diagnostic patterns, including electrophoretically normal, acute‐phase protein responses, polyclonal gammopathies, restricted polyclonal/oligoclonal gammopathies, paraproteinemias (monoclonal or biclonal gammopathies), and Bence‐Jones proteinurias are also reviewed using relevant case material. Cases in which immunofixation electrophoresis are particularly useful are highlighted, and methodologies to more accurately quantify serum monoclonal proteins (M‐proteins), monitoring tests commonly used in human medicine, are discussed. [ABSTRACT FROM AUTHOR]