Electrophoresis Patterns Research Articles

Raubasine-Induced Groove Binding in Salmon Testes DNA: Exploring the Structural Modulation, Antiglycation, and Antioxidant Properties.

As one of nature's most fundamental blueprints and due to its critical role in life processes, DNA has naturally become the cornerstone of numerous research efforts. One particularly intriguing area of study is understanding how small molecules interact with nucleic acids. In this study, we investigated the interaction between the plant-derived indole alkaloid Raubasine (Ajmalicine; AJM) and Salmon Testes (ST) DNA using biophysical and computational techniques. A hyperchromic shift in the fluorescence intensity indicated the effective binding of AJM to ST DNA. The binding constant was in the order of 105 M-1 with a single preferential binding mode. Thermodynamic analysis revealed that exothermic binding was driven by positive entropy and negative enthalpy. The salt-dependent fluorescence analysis indicates the involvement of nonpolyelectrolytic forces in the interaction. Studies of iodide quenching, urea denaturation, dye displacement, and molecular docking further support that AJM binds to ST DNA through groove binding. Structural perturbation of DNA was evident from circular dichroism. The stability of the AJM-DNA complex was confirmed by molecular dynamics simulations. Prolonged elevated blood glucose levels induce nonenzymatic glycation of DNA, resulting in DNA-AGE (advanced glycation end-products) formation and free radical production, which disrupts the DNA structure. We explored ST-DNA glycation and its suppression by AJM. DNA-AGEs in vitro were characterized using UV-vis and fluorescence spectroscopy. The inhibition of glycation by AJM was assessed through changes in AGEs fluorescence intensity, gel electrophoresis patterns, and antioxidant activity, highlighting its ability to target glycated sites or neutralize free radicals generated during glycation. Our findings reveal AJM's potential to prevent the formation of AGEs, which may offer promising avenues for targeted therapies against glycation-related diseases such as diabetes, neurodegeneration, and cancer.

Assessment of serum protein profile in sickle cell disease

Background: Sickle Cell disease (SCD) is known to be caused by a mutation in the beta-globin gene of the hemoglobin that affects the red blood cells (RBCs) and is passed down through generations. This study was carried out to determine the effect of sickling of RBC on serum protein profile in SCD individuals. Methods: A case-controlled study was carried out among 80 patients. They were forty-five sickle cell disease individuals (HbSS) attending Children Specialist Hospital Ilorin and thirty-five healthy controls (HbAA). The levels of total protein and albumin were determined spectrophotometrically and serum protein electrophoresis (SPE) was carried out using cellulose acetate electrophoresis. Results: Significant hypoalbuminemia and hypergammaglobulinemia were observed in SCD groups compared with controls. A higher proportion of the SCD group, 11(25%) had hypergammaglobulinemia and 21(49%) had hypoalbuminemia (P<0.05). Plasma protein electrophoresis in SCD patients shows an intense colour at the gamma region which was not seen in control. There was a significant relationship between the patterns of serum protein electrophoresis and the frequency of crisis. Conclusions: Sickle cell disease crisis and other associated underlying conditions may be prevented early through assessment of serum protein profile, especially SPE. Hypergammaglobulinemia and hypoalbuminemia may be associated with frequent episode of crisis.

Utility of Immunofixation as a Follow-up to Select Abnormal Serum Protein Electrophoresis Patterns and Suggestions for Clinical Correlation

Utility of Immunofixation as a Follow-up to Select Abnormal Serum Protein Electrophoresis Patterns and Suggestions for Clinical Correlation

Modifying physicochemical properties, rheology, and creaming stability of milk fat globule and membrane through ultrasound treatment

The healthy benefits of milk fat globules and membrane (MFGs/MFGM) ingredients are increasingly recognized in the dairy industry. In this research, we examined the effects of ultrasonic treatment on the physicochemical and rheological properties, as well as the emulsions stability of MFGs/MFGM derived from bovine raw milk. Fresh milk was subjected to sonication at frequencies of 20 kHz and 40 kHz, either individually or simultaneously, for durations of 5 min or 15 min, using work/rest cycles of 5 s on and 3 s off. Bovine milk, without any treatment, served as the control. Regardless of the intensity difference, ultrasonic treatment for 5 min resulted in more pronounced changes in the regions of Amide Ⅱ (1600–1500 cm−1), Amide Ⅲ (1500–1200 cm−1), and fingerprint region (1200–1900 cm−1) compared to both the 15 min treatments and control MFGs/MFGM. Principal component analysis (PCA) conducted on the entire spectra, as well as in the regions of Amide Ⅰ, Amide Ⅱ, and the fingerprint spectra, clustered the 5 min treatment distinctly from the control and MFGs/MFGM ultrasonically treated for 15 min. MFGs/MFGM samples following 20 kHz and 40 kHz synchronous treatment for 15 min exhibited lower absorbance bands at 1727–1726 cm−1, whereas a higher content at 1740 cm−1 was observed compared to control MFGs/MFGM. Additionally, a more significant reduction in the intramolecular β-sheet content in 20 + 40 kHz/ 15 min treatment was observed. According to the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) patterns, a diminished intensity of Periodic Acid Schiff 6/7 (PAS 6/7) bands was observed across all the MFGs/MFGM. Ultrasonic treatment retained more caseins while reducing the β –LG levels compared to the controls, enhancing the stability of MFGs/MFGM, except in MFGs/ MFGM subjected to 20 and 40 kHz simultaneously treated for 15 min. The irregular sphericity of fat globules was noted particularly in MFGs/MFGM treated at 20 kHz independently or in combination with 40 kHz for 15 min. According to the confocal laser scanning microscopy (CLSM), ultrasonic treatment facilitated the binding of caseins or whey proteins to the MFGs surface and induced flocculation of membrane proteins. Hierarchical cluster analysis (HCA) heat map further underscored the impact of ultrasonic treatments on the structural and compositional changes, as well as rheology and emulsions stability, of MFGs/MFGM.

A first study of meat-borne enterococci from butcher shops: prevalence, virulence characteristics, antibiotic resistance and clonal relationship.

IntroductionEnterococcus, which used to be thought of as a harmless commensal living in the digestive tract, has now become a highly resistant and highly contagious pathogen that makes nosocomial infections much more common. This study examined enterococci species and their antibiotic resistance phenotypes and genotypes and virulence gene content in Turkish ground beef samples. Methodology A total of 100 ground beef samples were analyzed between May 2020 and May 2021. The isolated strains were identified via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and confirmed using polymerase chain reaction (PCR) after which they were divided into several species using PCR and tested for antibiotic resistance against 19 antimicrobial agents using the disc diffusion method. The genetic similarity analysis, pulsed-field gel electrophoresis (PFGE) was performed. Results A total of 93 isolates in ground beef were identified, comprised of E. faecalis 72.04%; E. hirae- 11.82%; E. casseliflavous- 7.52%; E. faecium- 5.3%; E. gallinarium- 3.23%. The virulence genes observed in Enterococcus species were distributed as follows: gelE 88.1%, ace 53.7%, efaA 40.8%, asaI 19.3%, esp 6.4%, and cylA 1.07%. A high antibiotic resistance was recorded for tetracycline (43.01%), followed by ampicilin (17.2%), and chloramphenicol (13.9%). 17.2% of Enterococcus isolates were multidrug-resistant. The study determined the high prevalence of antibiotic resistance genes, specifically for tet(L) 10 (10.7%), aac(6')Ie-aph(2")-la 3 (3.2%), and ermB 3 (3.2%). The presence of efflux pump genes were identified in 74.1% of Enterococcus isolates. Genetic characterization of 67 E. faecalis isolates by PFGE revealed 41 pulsed-field gel electrophoresis (PFGE) patterns that were grouped into 15 clusters, which presented more than one strain with 100% similarity. Conclusion Isolates obtained from several areas and butchers had comparable patterns of PFGE, suggesting that the presence of circulating E. faecalis poses a potential public health concern in diverse districts. To mitigate the health hazards associated with the contamination of enterococci from raw to cooked meats, it is necessary to enhance the disinfection of butcheries, promote excellent hand hygiene among butchers, and implement appropriate meat storage and handling methods to prevent bacterial development.

Cytotoxicity and Apoptotic DNA Fragmentation Evaluation of Ethanolic Extract of Phyllanthus niruri L. in Vero and Primary Feline Testicular Cells

Phyllanthus niruri or Dukung Anak is native to Southeast Asia, particularly in Malaysia and has been used as traditional preparations and as health supplements. However, previous studies have found that P. niruri can impair the male reproductive functions. With booming population of stray animals within the community, a new controlling method that is more affordable and safer must be developed. This study was aimed to explore the safety of P. niruri as a male herbal contraceptive to be used in veterinary medicine as one of the non-surgical sterilisation methods. For the preliminary part of this ongoing research, we have investigated the cytotoxicity activity of P. niruri ethanolic extract on Vero and primary feline testicular cells (FTC) and DNA apoptotic activity in FTC. Phyllanthus niruri extract was prepared using cold maceration method in 50% ethanol. The FTC were prepared from feline testes obtained from post-routine castration from local private veterinary clinics, while Vero cells were obtained from the archived cells of Virology Laboratory, Faculty of Veterinary Medicine Universiti Malaysia Kelantan (FPV UMK). The MTT assay was performed to evaluate the cytotoxicity of these plant extracts on FTC and Vero cells. Apoptosis was assessed using DNA laddering assay on extracted DNA of the treated FTC cells and observed the DNA patterns in gel electrophoresis. The 50% cytotoxic concentration (CC50) of the ethanolic extract of P. niruri was a dose dependent. The DNA laddering assay revealed that the plant extract does not induce apoptosis in FTC. Therefore, the study outcomes indicated that the ethanolic extract of P. niruri is potentially safe for animal use. However, in vivo studies are required to confirm these findings.

The effect of polysaccharide type on dielectric barrier discharge (DBD) plasma glycosylation of sodium caseinate-part I: Physicochemical, structural and thermal properties

This study aimed to investigate the impact of polysaccharide type on the physicochemical, structural, and thermal properties of dielectric barrier discharge (DBD) plasma glycosylated sodium caseinate (SC). The polysaccharides Quince seed gum (QSG), carboxymethyl cellulose (CMC), and maltodextrin (MD) were mixed with SC and treated with DBD plasma at 18 kV for 10 min. The grafting degree, electrophoresis pattern, FTIR, XRD, carbonyl, sulfhydryl, and di-tyrosine content, FE-SEM, color, and thermal properties of SC and its polysaccharide mixtures before and after plasma treatment were analyzed. Results showed that the SC-QSG conjugate had the highest glycation degree and color change after plasma treatment. The SC-CMC and SC-QSG conjugates exhibited disappearance of distinct SC bands in electrophoresis pattern compared to SC. Also, significant changes in functional group and crystallinity were occurred in SC-CMC conjugate. Plasma treatment caused oxidation of SC, but the presence of polysaccharides offered protection against oxidation. The microstructure of SC was altered by mixing with polysaccharides and exposure to plasma. Also, the mixtures indicated higher thermal stability after plasma treatment. Results confirmed that the generation of protein-polysaccharide conjugates through DBD plasma technique was depended on with SC-MD conjugate unable to form through this method.

Dyslipidaemia in women with polycystic ovary syndrome referred to a teaching hospital in Cape Town, South Africa.

The polycystic ovary syndrome (PCOS) imparts health risks including dyslipidaemia, diabetes and cardiovascular disease that are amenable to lifestyle adjustment and/or medication. We describe dyslipidaemia in women referred to a gynaecological endocrine clinic. Clinical data and endocrine and lipoprotein investigations comprising fasting triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDLC) and calculated low density lipoprotein cholesterol (LDLC) were studied along with electrophoresis patterns of apolipoprotein B-containing lipoproteins. The 1721 participants comprised black, mixed ancestry, white and Indian individuals (9.8%, 83.2%, 5.8% and 1.2%, respectively). The mean ± standard deviation of the age, body mass index (BMI) and waist/hip ratio were 26.0 ± 5.9 years, 32.3 ± 8.3 kg/m2 and waist/hip ratio 0.88 ± 0.11, respectively. Overweight status (BMI 26-30 kg/m2) and obesity (BMI >30 kg/m2) involved 272 (15.8%) and 1010 (58.7%) individuals, respectively. Morbid obesity (BMI >40 kg/m2) was present in 309 (17.9%) individuals. The TG, TC, HDLC and LDLC concentrations were 1.22 ± 0.86, 4.77 ± 1.02, 1.3 ± 0.36, 2.94 ± 0.94 mmol/L, respectively. LDL hypercholesterolaemia occurred in 753 (43.7%) and exceeded 5 mmol/L in 39 (2.3%) women. Low HDLC (<0.9 mmol/L) affected 122 (7%), hypertriglyceridaemia (>1.7 mmol/L) affected 265 (15.4%) and exceeded 2.5 mmol/L in 91 (5.3%) women. Mixed hyperlipidaemia (TG >1.7, TC >5.0 mmol/L) occurred in 176 (10.2%). Electrophoresis revealed small LDL particles in 79 (4.6%) and dysbetalipoproteinaemia in 13 (0.76%) of the cohort. Small LDL associated with obesity, blood pressure, TG and glucose concentration and higher androgenic state. Many women with PCOS had unfavourable lipoprotein results: mostly moderate changes in TG, HDLC and LDLC. Small LDL is not rare, may aid risk assessment and is best determined directly. Incidental monogenic disorders of lipoprotein metabolism included dysbetalipoproteinaemia, familial hypercholesterolaemia and severe hypertriglyceridaemia. Dyslipidaemia in PCOS requires more careful diagnosis, individualised management and research.

Method of Immunoglobulin Y (IgY) Extraction for Minimally Toxic In Vivo Applications

Although immunoglobulin (IgY) has therapeutic potential, extracts from conventional methods are generally parenterally incompatible. This present study aimed to design a method to generate an IgY composition in vivo administration. The proposed procedure incorporated the following modifications: delipidation using carrageenan, salt precipitation using sodium sulfate, desalting using gel filtration, and affinity chromatography using MgCl2 as a disrupting agent. Furthermore, specific IgY was generated against human histones (H3 or H4) or calf histones to demonstrate the maintenance of target specificity. Native polyacrylamide gel electrophoresis patterns showed increasing purity from crude to post-desalted samples. Affinity chromatography estimated specific antibodies in the crude extract at 3–8%. Using ELISA, specificity was demonstrated for plasma, serum, or bicarbonate buffer samples. No mortality was observed in Sprague-Dawley rats and BALB/c mice using OECD up-and-down limit protocol adjusted to wrote. LD50 was estimated at more than 500 mg/kg intraperitoneally and 300 mg/kg intravenously. Intravenously injected with limited doses, intraperitoneally in mice, and daily sublethal doses for up to 36 d. Curiously, nonspecific IgY – but not specific IgY – had a higher liver injury score than solvent alone for acute injections (P &lt; 0.05). Thus, the proposed method of IgY isolation may offer a means for an IgY composition that can be compatible with both in vivo therapeutics.

Development of Loop-Mediated Isothermal Amplification Assays for the Rapid and Accurate Diagnosis of Exserohilum turcicum for Field Applications.

Northern corn leaf blight (NCLB), caused by Exserohilum turcicum, is one of the most devastating foliar diseases of maize. Rapid and accurate diagnosis for this disease is urgently needed but still limited. Here, we establish a field-deployable diagnostic method to detect E. turcicum based on loop-mediated isothermal amplification (LAMP) assays. A software application called K-mer Elimination by Cross-reference was used to search for the specific sequences belonging to E. turcicum by comparing the whole genome sequence between E. turcicum and other known maize pathogens. Five LAMP primer sets were designed based on specific and single-copy fragments of E. turcicum. Post-LAMP analyses indicated that only the primer set, Et9468_set1, was the most suitable, producing a ladder-like amplification pattern in the agarose gel electrophoresis and a strong fluorescence signal in the presence of SYBR Green I. The LAMP assay using Et9468_set1 primers demonstrated a high level of specificity in distinguishing E. turcicum from six other common fungal pathogens of maize, as well as 12 more fungal and oomycete strains including the epiphytic fungi from maize leaves and other crop pathogens. Moreover, it exhibited remarkable sensitivity by detecting five copies per reaction, which was approximately 104 times more sensitive compared with conventional PCR. The LAMP assay successfully detected E. turcicum in field maize leaves without DNA extraction, demonstrating its suitability for rapid on-spot detection of NCLB. Our study provides a direct LAMP diagnostic method to detect E. turcicum, which enables on-site pathogen detection in the field and the development of preventive strategies for NCLB management.

Different cellular and molecular responses of Bovine milk phagocytes to persistent and transient strains of Streptococcus uberis causing mastitis.

Streptococcus uberis is frequently isolated from milk collected from dairy cows with mastitis. According to the host's immunity, bacterial virulence, and their interaction, infection with some strains can induce persistent subclinical inflammation, while infection with others induces severe inflammation and transient mastitis. This study compared the inflammatory response of milk-isolated white blood cells (mWBCs) to persistent and transient S. uberis strains. Quarter milk samples were collected aseptically for bacterial culture from all lactating cows once a week over a 10-week period. A transient and noncapsular strain with a 1-week intramammary infection duration was selected from this herd, while a persistent and capsular S. uberis strain with an intramammary infection longer than 2 months from our previous study was selected based on an identical pulse field gel electrophoresis pattern during the IMI episode. Cellular and molecular responses of mWBCs were tested, and the data were analyzed using repeated analysis of variance. The results showed a higher response in migration, reactive oxygen species generation, and bacterial killing when cells were stimulated with transient S. uberis. In contrast, the persistent strain led to increased neutrophil extracellular trap release. This study also highlighted several important molecular aspects of mWBCs. Gene expression analyses by real-time RT-PCR revealed a significant elevation in the expression of Toll-like receptors (TLR-1, TLR-2, TLR-6) and proinflammatory cytokines (tumor necrosis factor-alpha or TNF-α) with the transient strain. Additionally, Streptococcus uberis capsule formation might contribute to the capability of these strains to induce different immune responses. Altogether, these results focus on the immune function of activated mWBCs which demonstrate that a transient strain can elicit a stronger local immune response and, subsequently, lead to rapid recovery from mastitis.

Sickle Cell Gene Frequency Analysis in Visapur, District Chandrapur: A Case Study of Single Village from Central India

Sickle cell anemia burden is generally studied by calculating how many sickle cell carriers are found in specific diagnostic camps based on combination of screening test and confirmatory tests i.e. sickle cell carrier prevalence method. Where as actual total numbers of alleles can be estimated by combining data of how many carriers and homozygous patients are found in the confirmatory testing camps, which is a gene frequency calculation method. Aim of this study is to compare prevalence and gene frequency calculations for one village data, for study conducted in year 2005 at Visapur village, Chandrapur district, Central India. Sampling was done in village school followed by screening and confirmatory testing at field work laboratory at Ballarpur city which included solubility test, hemoglobin estimation, hemoglobin electrophoresis using cellulose acetate membrane method followed by pedigree analysis after parent studies. This is a sickle cell anaemia study of single village data point. Out of total population of the village, 6.68% population were randomly studied. Total registrations done for 755 individuals and out of that total 628 individuals were tested (N = 628, 293 were females and 335 were males) for solubility tests followed by haemoglobin electrophoresis. For socioeconomically disadvantages groups, it was noted that out of 628 individuals, tribals were 111 and non-tribals, 517. Out of total tested, N=628, SCD homozygous status were found in 7 cases (Hb S+S electrophoresis pattern). In 87 cases sickle cell carrier status (Hb A+S electrophoresis pattern) were observed and in remaining 534 individuals, non-sickle cell status observed i.e. Hb A+A electrophoresis pattern. Based on this sickle cell prevalence observed for total tribal and non-tribal is 13.85% After classifying tribal Vs non-tribal population, 7 homozygous patients were found in non-tribal communities, 5 in Schedule caste, 1 in OBC and 1 in open category, there is no SCD patient found in tribal community. Out of 50 sickle cell carriers, 7 were found from tribal community and remaining all 80 belongs to non-tribal community. Out of which, 49 are from scheduled cast and 31 from OBC community. After studying sickle cell prevalence and gene frequency pattern, it is found that gene frequency gives more accurate status of sickle cell anaemia even in small population size, i.e. independent study of single village, as compared to estimation sickle cell anemia using sickle cell carrier prevalence method.

Pattern of ABO blood group and haemoglobin electrophoresis (phenotype) among patients presenting with acute Noma in a referral center, Northwest Nigeria

Abstract Background: Noma is a necrotizing destructive disease of the orofacial structures with poorly understood etiopathogenesis. This study aimed to present the pattern of ABO blood group and hemoglobin electrophoresis (phenotype) in patients presenting with acute Noma. Materials and Methods: A retrospective study conducted over five months (November, 2023 to April, 2024) using five year record of patients who presented with acute noma. After obtaining ethical approval, case notes of patients who were managed for acute Noma were retrieved. Patients’ demographic information such as age and sex was recorded. The blood group and the hemoglobin phenotype of the patients were also recorded. Data obtained were analyzed using the Statistical Package for the Social Sciences (IBM SPSS version 25). Results: A total of 205 subjects were recruited, of which there were 100 (48.8%) males and 105 (51.2%) females. The age mean ± standard deviation of the patient was 5.06 ± 3.74 in the range of 9 months–22 years. The blood group O 119 (58.3%) was the highest. The majority of the subjects 179 (87.3%) were found to have rhesus positive blood group antigen. Blood group O was found to have the highest positive rhesus antigen with a significant difference (χ 2 = 9.620, df = 3, P = 0.022). The association between the ABO blood group and the phenotype showed that subjects with blood group O have the highest AA phenotype with no significant difference (χ 2 = 8.855, df = 6, P = 0.182). Conclusion: This study revealed more frequent blood group “O” rhesus positive and “AA” hemoglobin electrophoresis (phenotype) in patients with acute Noma which is similar to the general population in the same geographical region.

Gel mechanism analysis of minced scallop (Patinopecten yessoensis) meat modified by three kinds of food colloids

Scallop is a potential resource of gel products. But its application is limited due to its bad gelation. The effect of different food hydrocolloids (curdlan, κ-carrageenan and pectin) was investigated on the gel properties, moisture distribution and gel formation. Curdlan and κ-carrageenan increased the gel strength to 2.13 N and 2.09 N and the hardness to 3.29 N and 3.12 N respectively by binding with myofibrillar protein and reducing the water mobility while pectin was detrimental to gelation of scallop mince due to its competition with water molecule. Curdlan and κ-carrageenan could increase the gelation by promoting the formation of disulfide bonds (11.1134 mg/mL) and hydrophobic bonds (2.8904 mg/mL) in the scallop minced network, respectively. Scanning electron microscopy (SEM) showed that the network structure of curdlan and κ-carrageenan was more dense and compact, while pectin was the opposite. The SDS-PAGE electrophoresis pattern showed that curdlan and κ-carrageenan caused a certain degree of aggregation of the sample proteins. Therefore, curdlan and κ-carrageenan could effectively improve the gel properties of scallop products, while pectin has the opposite effect. These results provide a basis for further understanding the effect of food colloid on the gel properties of scallop protein.

Simultaneous probing of transcription, G-quadruplex, and R-loop.

DNA and RNA can form various non-canonical secondary structures, including G-quadruplex (G4) and R-loops. These structures are considered transcriptional regulatory elements due to their enrichment at regulatory regions. During transcription, G-rich sequences in the non-template strand promote R-loop formation in the DNA template strand. These R-loops induce G4 structures in the non-template DNA strand, further stabilizing them. Additionally, the high rG: dC base-pairing within the R-loop contributes to the stability of DNA/RNA hybridization. Our previous study investigated the interplay between G4s and R-loops and its impact on transcription. We employed two techniques to demonstrate transcription-mediated G4 and R-loop formation. The single-molecule method allows us to detect intricate details of transcription initiation, elongation, and co-transcriptional R-loop and G4 formation. It provides a high-resolution view of the dynamic processes involved in transcriptional regulation. As an orthogonal approach, a gel-based assay enables the detection of the transcription-mediated R-loops and the RNA product. We can measure the progressive formation of R-loop and total RNA produced from transcription by analyzing gel electrophoresis patterns. In summary, these techniques provide valuable insights into the non-canonical nucleic acid structures and their impact on gene expression.

Carbapenemase-producing Enterobacterales isolated from hospital sinks: molecular relationships with isolates from patients and the change in contamination status after daily disinfection with sodium hypochlorite.

This study aimed to investigate the contamination status of hospital sinks with carbapenemase-producing Enterobacterales (CPE), the efficacy of daily cleaning with sodium hypochlorite, and the relationships between CPEs isolated from contaminated sinks and patients. Pre/postintervention surveys of the CPE-contaminated sinks. Hospital wards including pediatric intensive care unit in a children's hospital. Consenting CPE-colonized patients admitted between November 2018 and June 2021 in our hospital. Environmental culture of 180 sinks from nine wards in our hospital was performed three times with an interval of 2 years (2019, 2021, 2023). Molecular typing of the isolated strains from the sinks and patients was performed. After the first surveillance culture, we initiated daily disinfection of the sinks using sodium hypochlorite. Before the intervention, we detected 30 CPE-positive sinks in 2019. After the intervention with sodium hypochlorite, we observed a substantial decline in the number of sinks contaminated with CPE; 13 in 2021 and 6 in 2023. However, the intervention did not significantly reduce the number of CPE-contaminated sinks used for the disposal of nutrition-rich substances. The CPE isolates from the patients and those from the sinks of the wards or floors where they were admitted tended to have similar pulse-field gel electrophoresis patterns. Contaminated sinks could be reservoirs of disseminating CPE to the patients. Daily disinfection of sinks with sodium hypochlorite may be effective in eliminating CPE, although the effect could be weaker in sinks with a greater risk of contact with nutrition-rich substances.

Antioxidant properties of bovine liver protein hydrolysates and their practical application in biphasic systems.

The influence of protein hydrolysate produced from bovine liver protein hydrolysate (LPH) by enzymatic hydrolysis, using Alcalase/Protamex (1:1), on lipid dispersions was investigated. LPH production was optimized to maximize the antioxidant activity (at 45, 50, and 55 °C for 12, 18, and 24 h). Different concentrations of LPHs (1, 3, and 5 mg/g) were added to emulsions and to liposomes. Lipid oxidation level and particle size of the lipid dispersions were monitored for 14 days of storage at 25 °C. Radical scavenging activity and reducing power were the highest at 45 °C after 24 h of hydrolysis. Electrophoresis pattern showed that the antioxidant activity was arising from the peptides with molecular weight around 10 kDa. Lipid oxidation occurred more rapidly in samples without LPH during storage. In emulsions, lower thiobarbituric acid-reactive substance and conjugated diene values were measured with increasing concentrations of LPH at day 14. Accordingly, particle size of the samples containing 5 mg/g of LPH was smaller than those of other groups. Phase separation was observed only in lecithin emulsion without LPH at day 14. The use of LPH in liposome limited the lipid oxidation and maintained the size of the particles independently from the concentration. This study highlights the potential applications of animal by-products as natural antioxidants in complex food systems. The results demonstrate that LPH, particularly when hydrolyzed at optimized conditions, can effectively inhibit lipid oxidation. The findings suggest that biphasic systems incorporating LPH have promising prospects for enhancing the stability and quality of food products. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Investigation of some changes and clonal relationship in enterococci isolates due to relocation of a hospital.

To investigate the isolation rates, antimicrobial resistance rates, minimum inhibitory concentration values of antimicrobial agents, and clonal relationships of Enterococcus faecalis and Enterococcus faeciumdue to the relocation of a hospital to a newly constructed building. The comparative, prospective study was conducted at adult general intensive care units of the Mus State Hospital, Mus, Turkey, in two phases; before the relocation from January 25 to December 1, 2014, and after the relocation from February 10 to May 24, 2015. Rectal swab samples were collected 72 hours post-hospitalisation. Identification of Enterococcus faecalis and Enterococcus faeciumisolates was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and antimicrobial resistance with minimum inhibitory concentration values was detected with Vitek 2 system. The clonal relatedness among the strains was investigated by pulsed-field gel electrophoresis. Data was analysed using SPSS 23. Of the 69 patients, 37(53.62%) were related to pre-relocation phase; 20(54.1%) females and 17(45.9%) males with mean age 62.81±21.71 years. There were 32(46.37%) patients in the post-relocation phase; 13(40.6%) females and 19(59.4%) males with mean age 62.69±21.35 years (p>0.05). Of the 84 enterococci strains isolated, 51(60.7%) were Enterococcus faecium; 28(55%) before relocation and 23(45%) after relocation (p=0.77). The remaining 33(39.3%) isolates were Enterococcus faecalis; 16(48.5%) before relocation and 17(51.5%) after relocation (p=0.73). Multiple strains were located in 7(18.9%) patients before relocation and in 7(21.9%) after relocation. In 1(3.1%) patient after relocation, 2(8.7%) Enterococcus faecium isolates with different resistance and pulsed-field gel electrophoresis patterns were detected. There were no significant differences between the isolation and antibiotic resistance rates before and after relocation (p>0.05), and a clonal relation between the isolates was not detected (p>0.05). Decreased minimum inhibitory concentration values were noted for some antibiotics. Clonal relationship between the isolates and change in the rates of isolation and antimicrobial resistance of Enterococcus faecalis and Enterococcus faecium was not detected due to relocation. Minimum inhibitory concentration values could be used to reveal relocation-related changes in isolates obtained from patients hospitalised in intensive care units.

Presence of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Food-Producing and Companion Animals and Wildlife on Small-Holder Farms of Floreana Island, Galápagos Islands.

Background: Antimicrobial resistance (AR) has led to increasing human and animal morbidity and mortality and negative consequences for the environment. AR among Escherichia coli (EC) is on the rise, with serious concerns about extended-spectrum β-lactamase-producing E. coli (ESBL-EC). In the Galápagos Islands, where antimicrobials are available without a prescription, growing demands for food production can drive antimicrobial use. Food producing animals are at the interface of wildlife and environmental health on the smallest human-inhabited Galápagos Island, Floreana. We sought to determine if ESBL-EC were present in Floreana Island farm animal species and nearby wildlife and the relatedness of ESBL-EC isolates identified. Materials and Methods: During July 4-5, 2022, we visited 8 multispecies farms, representing 75% of food-producing animal production on Floreana, and collected 227 fecal samples from farm animals and wildlife. Each sample was plated on MacConkey agar supplemented with cefotaxime (4 μg/mL). Results: ESBL-EC was isolated from 20 (9%) fecal samples collected from pigs (N = 10), chickens (N = 6), wildlife (N = 3), and dog (N = 1). All ESBL-EC isolates were from samples taken at three (38%) of the eight farms. Fifteen (75%) of the ESBL-EC isolates were from a single farm. All ESBL-EC isolates were multidrug resistant. The most prevalent ESBL genes belonged to the blaCTX-M group. Among the typeable isolates from the farm with the largest proportion of ESBL-EC isolates (N = 14), we observed nine unique pulsed-field gel electrophoresis (PFGE) patterns, with identical patterns present across pig and chicken isolates. PFGE patterns in the three farms with ESBL-EC isolates were different. Conclusions: These results lend support for future routine AR monitoring activities at the livestock-wildlife interface in Galápagos to characterize potential interspecies transmission of AR bacteria and AR genes in this unique protected ecosystem, and the related human, animal, and environmental health impacts, and to formulate interventions to reduce AR spread in this setting.

Retrospective study on the distribution of hemoglobinopathies in Karnataka-A laboratory experience.

The importance of screening for hemoglobinopathies is well-documented in India. However, information on the distribution of hemoglobinopathies in Karnataka is lacking. The present study focuses on determining the spectrum of hemoglobinopathies for various districts of Karnataka. A retrospective analysis of samples registered for hemoglobinopathies for a period of 5 years (2017-2021) was carried out. A total of 17066 records registered only from the Karnataka region, were anonymized and retrieved. The data included gender, age, district, and results of the tests. The results were based on complete blood count, peripheral smear, and capillary electrophoresis (CE) pattern. The data were revalidated by pathologists, and the unambiguous data were analyzed for the study. One-fourth of the records (25%) showed abnormal hematological parameters. The number of female records (66%) was twice that of males and both genders showed higher distribution of thalassemia, followed by variants and double heterozygotes (DH). Several cases of thalassemia major were identified below the age of 17 years. The majority of thalassemia cases were β thal and 93% of them were β thal trait. Among the variants, HbS was more prevalent than HbE. Among the districts, Hassan had a 35.2% thal, Mysuru had a 7.2% variant, and Chitradurga had a 5.5% DH. Thalassemia, variants, and DH were distributed across several districts of Karnataka to various levels. The comprehensive retrospective analysis of the spectrum of hemoglobinopathies in various districts of Karnataka serves as evidence to carry out a prospective study on population screening where the incidence of thalassemia and structural variants is high.