Day Treatment Research Articles

Latrophilin-2 Deletion in Cardiomyocyte Disrupts Cell Junction, Leading to D-CMP.

Latrophilin-2 (Lphn2), an adhesive GPCR (G protein-coupled receptor), was found to be a specific marker of cardiac progenitors during the differentiation of pluripotent stem cells into cardiomyocytes or during embryonic heart development in our previous studies. Its role in adult heart physiology, however, remains unclear. The embryonic lethality resulting from Lphn2 deletion necessitates the establishment of cardiomyocyte-specific, tamoxifen-inducible Lphn2 knockout mice, which was achieved by crossing Lphn2flox/flox mice with mice having MerCreMer (tamoxifen-inducible Cre recombinase) under the α-myosin heavy chain promoter. Tamoxifen treatment for several days completely suppressed Lphn2 expression, specifically in the myocardium, and induced the dilated cardiomyopathy (D-CMP) phenotype with serious arrhythmia and sudden death in a short period of time. Transmission electron microscopy showed mitochondrial abnormalities, blurred Z-discs, and dehiscent myofibrils. The D-CMP phenotype, or heart failure, worsened during myocardial infarction. In a mechanistic study of D-CMP, Lphn2 knockout suppressed PGC-1α and mitochondrial dysfunction, leading to the accumulation of reactive oxygen species and the global suppression of junctional molecules, such as N-cadherin (adherens junction), DSC-2 (desmocollin-2; desmosome), and connexin-43 (gap junction), leading to the dehiscence of cardiac myofibers and serious arrhythmia. In an experimental therapeutic trial, activators of p38-MAPK, which is a downstream signaling molecule of Lphn2, remarkably rescued the D-CMP phenotype of Lphn2 knockout in the heart by restoring PGC-1α and mitochondrial function and recovering global junctional proteins. Lphn2 is a critical regulator of heart integrity by controlling mitochondrial functions and cell-to-cell junctions in cardiomyocytes. Its deficiency leads to D-CMP, which can be rescued by activators of the p38-MAPK pathway.

The effects of alternate-day fasting on sleep and physical activity in poor sleeping adults: A randomized control trial.

Utilizing a randomized control design, 42 healthy adults (22.5 ± 2.8 years) participated in alternate-day modified fasting over a 12-day treatment period. Assessments of sleep included sleep time, efficiency, latency and wake after sleep onset, and assessments of physical activity included steps, energy expenditure, sedentary time, time spent in light physical activity and time spent in moderate-to-vigorous activity. Additional measurements included body composition and mood. The alternate-day modified fasting group consumed 25.8% ± 0.3% fewer calories compared with the control group (p = 0.03). There were no differences between groups for change in body mass index (p = 0.87), total fat mass (p = 0.91) or total lean mass (p = 0.88). Daily energy expenditure did not differ between groups (p = 0.11). On fast days, participants spent 34.5 ± 12.7 more minutes sedentary (p = 0.01), took 1100 ± 362 fewer steps (p < 0.01), and engaged in 27.2 ± 8.4 fewer minutes of moderate-to-vigorous physical activity (p = 0.00) compared with non-fasting days. Sleep duration, efficiency, latency or wake after sleep onset were not different between conditions (p = 0.92, p = 0.10, p = 0.09 and p = 0.66, respectively). We conclude that alternate-day modified fasting does not alter sleep time, efficiency, latency or wake after sleep onset in people reporting poor sleep quality, and does not alter overall physical activity. Although average daily physical activity is not altered, fasting in this manner does tend to result in more sedentary time and less physical activity with compensation on non-fasting days.

Transcriptomic analysis reveals the regulatory mechanisms of messenger RNA (mRNA) and long non-coding RNA (lncRNA) in response to waterlogging stress in rye (Secale cereale L.)

BackgroundWaterlogging stress (WS) negatively impacts crop growth and productivity, making it important to understand crop resistance processes and discover useful WS resistance genes. In this study, rye cultivars and wild rye species were subjected to 12-day WS treatment, and the cultivar Secale cereale L. Imperil showed higher tolerance. Whole transcriptome sequencing was performed on this cultivar to identify differentially expressed (DE) messenger RNAs (DE-mRNAs) and long non-coding RNAs (DE-lncRNAs) involved in WS response.ResultsAmong the 6 species, Secale cereale L. Imperil showed higher tolerance than wild rye species against WS. The cultivar effectively mitigated oxidative stress, and regulated hydrogen peroxide and superoxide anion. A total of 728 DE-mRNAs and 60 DE-lncRNAs were discovered. Among these, 318 DE-mRNAs and 32 DE-lncRNAs were upregulated, and 410 DE-mRNAs and 28 DE-lncRNAs were downregulated. GO enrichment analysis discovered metabolic processes, cellular processes, and single-organism processes as enriched biological processes (BP). For cellular components (CC), the enriched terms were membrane, membrane part, cell, and cell part. Enriched molecular functions (MF) terms were catalytic activity, binding, and transporter activity. LncRNA and mRNA regulatory processes were mainly related to MAPK signaling pathway-plant, plant hormone signal transduction, phenylpropanoid biosynthesis, anthocyanin biosynthesis, glutathione metabolism, ubiquitin-mediated proteolysis, ABC transporter, Cytochrome b6/f complex, secondary metabolite biosynthesis, and carotenoid biosynthesis pathways. The signalling of ethylene-related pathways was not mainly dependent on AP2/ERF and WRKY transcription factors (TF), but on other factors. Photosynthetic activity was active, and carotenoid levels increased in rye under WS. Sphingolipids, the cytochrome b6/f complex, and glutamate are involved in rye WS response. Sucrose transportation was not significantly inhibited, and sucrose breakdown occurs in rye under WS.ConclusionsThis study investigated the expression levels and regulatory functions of mRNAs and lncRNAs in 12-day waterlogged rye seedlings. The findings shed light on the genes that play a significant role in rye ability to withstand WS. The findings from this study will serve as a foundation for further investigations into the mRNA and lncRNA WS responses in rye.

Carbohydrate-Binding Properties and Antimicrobial and Anticancer Potential of a New Lectin from the Phloem Sap of Cucurbita pepo.

A Cucurbita phloem exudate lectin (CPL) from summer squash (Cucurbita pepo) fruits was isolated and its sugar-binding properties and biological activities were studied. The lectin was purified by affinity chromatography and the hemagglutination assay method was used to determine its pH, heat stability, metal-dependency and sugar specificity. Antimicrobial and anticancer activities were also studied by disc diffusion assays and in vivo and in vitro methods. The molecular weight of CPL was 30 ± 1 KDa and it was stable at different pH (5.0 to 9.0) and temperatures (30 to 60 °C). CPL recovered its hemagglutination activity in the presence of Ca2+. 4-nitrophenyl-α-D-glucopyranoside, lactose, rhamnose and N-acetyl-D-glucosamine strongly inhibited the activity. With an LC50 value of 265 µg/mL, CPL was moderately toxic and exhibited bacteriostatic, bactericidal and antibiofilm activities against different pathogenic bacteria. It also exhibited marked antifungal activity against Aspergillus niger and agglutinated A. flavus spores. In vivo antiproliferative activity against Ehrlich ascites carcinoma (EAC) cells in Swiss albino mice was observed when CPL exerted 36.44% and 66.66% growth inhibition at doses of 3.0 mg/kg/day and 6.0 mg/kg/day, respectively. A 12-day treatment by CPL could reverse their RBC and WBC counts as well as restore the hemoglobin percentage to normal levels. The MTT assay of CPL performed against human breast (MCF-7) and lung (A-549) cancer cell lines showed 29.53% and 18.30% of inhibitory activity at concentrations of 128 and 256 µg/mL, respectively.

Utilizing Disease Prediction Models to Time Fungicide Applications for Controlling Ripe Rot, Caused by Colletotrichum spp., in Maryland Vineyards.

Two previously published ripe rot prediction models, DF2-NN and GH2-DT, were evaluated for fungicide application timing efficacy in Maryland vineyards. Both models utilize leaf wetness duration (LWD), temperature, and grape cluster phenological stages as model parameters. These three parameters were tracked throughout the 2021 to 2023 seasons in three vineyards. The fungicide efficacy trials started at the veraison phenological stage and included a nontreated control, a 12-day interval treatment, and two model-triggered treatments when risk predicted by the models crossed a threshold. The severity of ripe rot on the clusters in each treatment was assessed when the fruit were mature. Ripe rot severity in the nontreated controls was higher during seasons with more LWD and more precipitation. Days in which the models were triggered by the environmental conditions primarily coincided with precipitation events and lengthy LWDs. The model-triggered treatments never had significantly higher ripe rot severity than the 12-day interval treatment but had significantly lower severities than the nontreated control in most trials which had high ripe rot pressure. Furthermore, the model-triggered treatments resulted in fewer fungicide applications than the 12-day interval treatment on average. The DF2-NN model appeared to be more accurate and useful for ripe rot prediction and treatment than the GH2-DT model because it triggered fewer fungicide applications while reducing ripe rot. This model may be useful for improving or maintaining ripe rot control with fewer fungicide inputs, which may be beneficial for the environment and the reduction of fungicide resistance selection.

Novel report on soil infection with Metarhizium rileyi against soil-dwelling life stages of insect pests.

Entomopathogenic fungi are the most effective control remedy against a wide range of medical and agricultural important pests. The present study aimed to isolate, identify, and assess the virulence of Metarhizium rileyi against Spodoptera litura and Spodoptera frugiperda pupae under soil conditions. The biotechnological methods were used to identify the isolate as M. rileyi. The fungal conidial pathogenicity (2.0 × 107, 2.0 × 108, 2.0 × 109, 2.0 × 1010, and 2.0 × 1011 conidia/mL-1) was tested against prepupae of S. litura and S. frugiperda at 3, 6, 9, and 12 days after treatments. Additionally, the artificial soil-conidial assay was performed on a nontarget species earthworm Eudrilus eugeniae, using M. rileyi conidia. The present results showed that the M. rileyi caused significant mortality rates in S. litura pupae (61-90%), and S. litura pupae were more susceptible than S. frugiperda pupae (46%-73%) at 12 day posttreatment. The LC50 and LC90 of M. rileyi against S. litura, were 3.4 × 1014-9.9 × 1017 conidia/mL-1 and 6.6 × 105-4.6 × 1014 conidia/mL-1 in S. frugiperda, respectively. The conidia of M. rileyi did not exhibit any sublethal effect on the adult stage of E. eugeniae, and Artemia salina following a 12-day treatment period. Moreover, in the histopathological evaluation no discernible harm was observed in the gut tissues of E. eugeniae, including the lumen and epithelial cells, as well as the muscles, setae, nucleus, mitochondria, and coelom. The present findings provide clear evidence that M. rileyi fungal conidia can be used as the foundation for the development of effective bio-insecticides to combat the pupae of S. litura and S. frugiperda agricultural pests.

Data-dependent and -independent acquisition lipidomics analysis reveals the tissue-dependent effect of metformin on lipid metabolism.

Despite the well-recognized health benefits, the mechanisms and site of action of metformin remains elusive. Metformin-induced global lipidomic changes in plasma of animal models and human subjects have been reported. However, there is a lack of systemic evaluation of metformin-induced lipidomic changes in different tissues. Metformin uptake requires active transporters such as organic cation transporters (OCTs), and hence, it is anticipated that metformin actions are tissue-dependent. In this study, we aim to characterize metformin effects in non-diabetic male mice with a special focus on lipidomics analysis. The findings from this study will help us to better understand the cell-autonomous (direct actions in target cells) or non-cell-autonomous (indirect actions in target cells) mechanisms of metformin and provide insights into the development of more potent yet safe drugs targeting a particular organ instead of systemic metabolism for metabolic regulations without major side effects. To characterize metformin-induced lipidomic alterations in different tissues of non-diabetic male mice and further identify lipids affected by metformin through cell-autonomous or systemic mechanisms based on the correlation between lipid alterations in tissues and the corresponding in-tissue metformin concentrations. A dual extraction method involving 80% methanol followed by MTBE (methyl tert-butyl ether) extraction enables the analysis of free fatty acids, polar metabolites, and lipids. Extracts from tissues and plasma of male mice treated with or without metformin in drinking water for 12days were analyzed using HILIC chromatography coupled to Q Exactive Plus mass spectrometer or reversed-phase liquid chromatography coupled to MS/MS scan workflow (hybrid mode) on LC-Orbitrap Exploris 480 mass spectrometer using biologically relevant lipids-containing inclusion list for data-independent acquisition (DIA), named as BRI-DIA workflow followed by data-dependent acquisition (DDA), to maximum the coverage of lipids and minimize the negative effect of stochasticity of precursor selection on experimental consistency and reproducibility. Lipidomics analysis of 6 mouse tissues and plasma allowed a systemic evaluation of lipidomic changes induced by metformin in different tissues. We observed that (1) the degrees of lipidomic changes induced by metformin treatment overly correlated with tissue concentrations of metformin; (2) the impact on lysophosphatidylcholine (lysoPC) and cardiolipins was positively correlated with tissue concentrations of metformin, while neutral lipids such as triglycerides did not correlate with the corresponding tissue metformin concentrations; (3) increase of intestinal tricarboxylic acid (TCA) cycle intermediates after metformin treatment. The data collected in this study from non-diabetic mice with 12-day metformin treatment suggest that the overall metabolic effect of metformin is positively correlated with tissue concentrations and the effect on individual lipid subclass is via both cell-autonomous mechanisms (cardiolipins and lysoPC) and non-cell-autonomous mechanisms (triglycerides).

Effect of the low-intensity pulsed ultrasound therapy on healing of Achilles tendinopathy in a rat model.

This study aimed to investigate the effect of the low-intensity pulsed ultrasound (LIPUS) on the healing of Achilles tendinopathy in a rat model induced by type 1 collagenase. The study was conducted on 144 Achilles tendons of 72 Wistar albino female rats with typical activity and weighing 300-350 g. The model of Achilles tendinopathy was created by injecting type 1 collagenase. According to the sampling time, 4 groups served as the control group, while 8 groups received treatment at varying periods. Low-intensity pulsed ultrasound therapy was initiated in 8 groups at 1, 7, and 15 days. Treatment was extended for 1 and 2 weeks. Achilles tendons were removed from the treatment and control groups on the 15th, 21st, 30th, and 45th days for biomechanical and pathologic examination. Compared to the control groups, LIPUS treatment administered in the first days of the proliferation phase increased tensile strength by approximately 30%, modulus of elasticity by approximately 53%, fibrillar appearance by 53%, and inflammation by 53%-33% in a shorter time. It was also demonstrated that starting treatment in the first days of the proliferation phase resulted in comparable success even with 1-week treatment compared to 2-week treatment. Low-intensity pulsed ultrasound therapy can provide positive results in managing Achilles tendinopathy in the rat model. Its capacity to shorten recuperation time has piqued the interest of conservative treatment approaches. As a result, more clinical research is required. Cite this article as:Kurtulmuş T, Çelebi ME, Bektaş E, Arican ÇD, Kucukyildirim BO, Demirkol M. Effect of the low-intensity pulsed ultrasound therapy on healing of Achilles tendinopathy in a rat model. Acta Orthop Traumatol Turc., 2024;58(2):102-109.

Retinoids in the treatment of photoageing: A histological study of topical retinoid efficacy in black skin.

Photoageing describes complex cutaneous changes that occur due to chronic exposure to solar ultraviolet radiation (UVR). The 'gold standard' for the treatment of photoaged white skin is all-trans retinoic acid (ATRA); however, cosmetic retinol (ROL) has also proven efficacious. Recent work has identified that black skin is susceptible to photoageing, characterized by disintegration of fibrillin-rich microfibrils (FRMs) at the dermal-epidermal junction (DEJ). However, the impact of topical retinoids for repair of black skin has not been well investigated. To determine the potential of retinoids to repair photoaged black skin. An exploratory intervention study was performed using an invivo, short-term patch test protocol. Healthy but photoaged black volunteers (>45 years) were recruited to the study, and participant extensor forearms were occluded with either 0.025% ATRA (n = 6; 4-day application due to irritancy) or ROL (12-day treatment protocol for a cosmetic) at concentrations of 0.3% (n = 6) or 1% (n = 6). Punch biopsies from occluded but untreated control sites and retinoid-treated sites were processed for histological analyses of epidermal characteristics, melanin distribution and dermal remodelling. Treatment with ATRA and ROL induced significant acanthosis (all p < 0.001) accompanied by a significant increase in keratinocyte proliferation (Ki67; all p < 0.01), dispersal of epidermal melanin and restoration of the FRMs at the DEJ (all p < 0.01), compared to untreated control. This study confirms that topical ATRA has utility for the repair of photoaged black skin and that ROL induces comparable effects on epidermal and dermal remodelling, albeit over a longer timeframe. The effects of topical retinoids on black photoaged skin are similar to those reported for white photoaged skin and suggest conserved biology in relation to repair of UVR-induced damage. Further investigation of topical retinoid efficacy in daily use is warranted for black skin.

Application of ultrasonography in diagnosis and treatment of children with congenital muscular torticollis.

Congenital muscular torticollis (CMT) is a more common childcare disease, which belongs to muscle-skeletal system diseases, and is more common in newborns. The disease is mainly due to congenital contracture due to chest locks, which leads to asymmetric head and neck. For such diseases, clear diagnosis and treatment in the early days is an important way to improve the prognosis of children. Compared with X-ray film, CT, and MRI, ultrasound examination has the advantages of low examination cost, short time, and no exposure to radiation during the examination. Moreover, ultrasound examination can provide an objective basis for the clinical diagnosis and prognosis evaluation of CMT children. This article reviews the latest research progress of conventional ultrasound, color Doppler ultrasound, and ultrasound elastography in the clinical diagnosis of CMT children and assisting in the formulation of treatment plans.

Safety and Tolerability of KIO-101 Eye Drops in Healthy Volunteers and Patients with Ocular Surface Disease-A Phase I Study.

Inhibitors of dihydroorotate dehydrogenase (DHODH) have been found to be potent anti-inflammatory agents. Recently, a topical formulation (KIO-101 eye drops) of a DHODH inhibitor has been developed. The aim of the present study was to evaluate the safety and tolerability of KIO-101 eye drops in Healthy Volunteers (HVs) and patients with conjunctival hyperemia. The study was carried out in a double-masked, placebo-controlled, randomized, parallel-group design with two parts. In part I, HVs received single and multiple instillations (four times daily for 12 consecutive days) of KIO-101 eye drops in ascending doses of 0.05%, 0.15%, and 0.30%, respectively. Part II was conducted in patients with conjunctival hyperemia who received 0.15% KIO-101 eye drops twice daily for 12 consecutive days. Ophthalmic and systemic safety examinations were performed on all participants. In part II, ocular hyperemia grading and an ocular surface disease index (OSDI) questionnaire were performed. 24 HVs participated in part I and 21 patients in part II. KIO-101 eye drops were well tolerated in all subjects. No serious adverse events (SAEs) occurred, and all AEs that were reported were transient and considered mild to moderate. In the highest dose cohort (0.30%), epistaxis occurred in two subjects after multiple instillations. In part II, after 12 days treatment with 0.15% KIO-101, conjunctival hyperemia decreased by -1.1 ± 0.27 points in the treatment and -0.6 ± 0.79 points in the placebo group (p = 0.0385). OSDI decreased from 47.9 ± 18.7 to 27.6 ± 19.13 points in the treatment group, while in the placebo group, a change from 41.3 ± 12.08 to 27.3 ± 18.63 points occurred. A 12-day treatment regimen with topical KIO-101 eye drops at low and mid doses was safe and well tolerated in both HVs and patients with conjunctival hyperemia. The obtained results point towards an early sign of reduction in conjunctival hyperemia.

G-ACP: a machine learning approach to the prediction of therapeutic peptides for gastric cancer

Conventional Gastrointestinal (GI) cancer treatments are quite expensive and have major hazards. Nowadays, a different strategy places more emphasis on creating tiny biologically active peptides that do not cause severe poisoning. Anticancer peptides (ACPs) are found through experimental screening, which is time-dependent and frequently fraught with difficulties. Gastric ACPs are emerging as a promising GI cancer treatment in the current day. It is crucial to identify novel gastric ACPs to have an improved knowledge of their functioning processes and treatment of gastric cancer. As a result of the post-genomic era’s massive production of peptide sequences, rapid and effective ACPs using a computational method are essential. Several adaptive statistical techniques for distinguishing ACPs and non-ACPs have recently been developed. A variety of adapted statistically significant methods have been developed to differentiate between ACPs and non-ACPs. Despite significant progress, there is no specific model for the prediction of gastric ACPs because the specific model will predict a particular type of peptide more accurately and quickly. To overcome this, an initiative is taken for the creation of a reliable framework for the accurate identification of gastric ACPs. The current technique in particular contains four possible features along with one hybrid feature encoding mechanisms which are the target-class motif previously indicated by Amino Acid Composition, Dipeptide Composition, Tripeptide Composition (TPC), Pseudo Amino Acid Composition (PAAC), and their Hybrid. Machine Learning algorithms make high-performance and accurate prediction tools. Moreover, highly variable and ideal deep feature selection is done using an ANOVA-based F score for feature pruning. Experiments on a range of algorithms are carried out to identify the optimal operating strategy due to the diverse nature of learning. Following analysis of the empirical results, Naïve Bayes with TPC and Hybrid feature space outperforms other methods with 0.99 accuracy score on the testing dataset. To find the model generalization an external validation is carried out. In external datasets, the Extra Trees with PAAC features outperforms with the accuracy of 0.94. The comparison study shows that our suggested model will predict gastric ACPs more accurately and will be useful in drug development and gastric cancer. The predictive model can be freely accessed at https://github.com/humeraazad10/G-ACP.git. Communicated by Ramaswamy H. Sarma

Surgical outcome after thyroidectomy due to Graves’ disease and Lugol iodine treatment: a retrospective register-based cohort study

PurposeThis study aimed to investigate the relationship between Lugol iodine treatment in a rescue setting and surgical outcomes in Graves’ disease patients.MethodsThe retrospective register-based cohort study included 813 patients who had undergone primary total thyroidectomy with a primary diagnosis of Graves’ disease (ICD-code E05.0) at Karolinska University Hospital in Stockholm, Sweden, between January 2008 and December 2015. Of 813 patients, 33 (4.1%) were given Lugol iodine before surgery and the remaining, the non-Lugol group, did not. The study’s primary outcomes were post-operative calcium treatment day 1, calcium and vitamin D supplements at discharge and follow-up. Secondary outcomes were laryngeal nerve damage and bleeding (defined as re-operation).ResultsDifferences were found between the Lugol and non-Lugol groups in the treatment of calcium day 1 (45.5% vs 26.7%, p = 0.018), at discharge (36.4% vs. 16.2%, p = 0.002) and vitamin D supplements at discharge (36.4% vs. 19.1%, p = 0.015) as surrogate variables for hypocalcemia post-operatively. No differences could be seen at 4–6 weeks and six-months follow-up. There were no differences between the Lugol and non-Lugol groups in terms of operation time, laryngeal nerve damage, and bleeding.ConclusionPatients in our cohort undergoing thyroidectomy due to Graves’ disease pre-operatively treated with Lugol iodine as a rescue therapy had a higher risk of experiencing short term post-operative hypocalcemia.

Unusual Location: Omental Ectopic Pregnancy Killing Me Slowly

Objective: Omentum pregnancy is an ectopic pregnancy in the abdominal cavity. The event has high morbidity and mortality. The purpose of this case is to present a rare case of abdominal ectopic pregnancy that occurred in a young woman.&#x0D; Methods: Case report.&#x0D; Results: A 22-year-old woman at 16 weeks gestation presented with abdominal pain and clinical shock. Examination results revealed pregnancy outside the womb, with an estimated fetal weight of 193 grams and positive heart activity. The patient underwent laparotomy surgery, revealing that the pregnancy had occurred in the omentum organ with placental attachment.&#x0D; Conclusion: In this case, emergency management began with the patient's reception in the emergency department, followed by laparotomy exploration. After a meticulous surgery, the pregnancy's location was identified in the omentum, and the evacuation of pregnancy products was performed. The patient received treatment for several days until being discharged home. Early diagnosis and interprofessional management are crucial if similar conditions are suspected in the future to prevent morbidity.&#x0D; Keywords: abdominal cavity, ectopic pregnancy, omental pregnancy

Serum pregnancy-associated glycoprotein profiles during early gestation in Karya and Konya Merino sheep.

Reported as being expressed by mono- and binucleate placental cells, pregnancy-associated glycoproteins (PAGs) are released into the blood circulation from the ruminant placenta. Circulating gestational PAGs levels may differ between sheep breeds. This study was aimed at the close monitoring of the serum PAGs profiles of Karya and Konya Merino sheep during early pregnancy. Fifteen Karya and 15 Konya Merino ewes were synchronized by a 12-day treatment with progesterone-impregnated intravaginal sponges. After the sponges were withdrawn, the ewes were administered 400IU of equine chorionic gonadotropin. The ewes were allowed to mate naturally, and all animals were sampled for blood as of the day of mating (day 0) at weeks 1, 2, 3, 4 and 5. Pregnancy diagnoses were made by transabdominal ultrasonography at week 5. At weeks 6, 7, 8, 9 and 11, blood samples were collected only from the pregnant ewes. The blood samples were centrifuged at 3000×g, and extracted sera were stored at -20°C until being used for laboratory analyses. Serum PAGs levels were determined with the aid of a commercial PAG-enzyme-linked immunosorbent assay test originally developed for pregnancy diagnosis in cattle. Differences in the between the PAGs levels throughout pregnancy and the group effect (Karya and Konya Merino) were determined with a two-way mixed analysis of variance. Pairwise comparisons were made using a Bonferroni adjustment. PAGs levels showed a linear increase with the advance of pregnancy in both Karya and Konya Merino sheep. No difference was detected between the breeds for serum PAGs levels. The serum PAGs levels of the pregnant and non-pregnant ewes differed as of the fourth week. The serum PAGs levels of the Karya and Konya Merino ewes were similar during the first 11 weeks of gestation, and pregnancy diagnosis could be made based on serum PAGs levels as of the 4th week in both breeds.

697. Characteristics of Hospitalized Patients Colonized with Clostridioides difficile at an Academic Medical Center

Abstract Background Clinical spectrum of C. difficile can range from colonization to fulminant colitis. Patients with toxin antigen negative, C. difficile nucleic acid amplification (NAAT)-positive diarrhea are frequently only colonized and do not suffer consequences of C. difficile associated disease (CDAD) but receive treatment for CDAD due to the perceived low risk of oral vancomycin vs. the risk of developing CDAD. Methods We performed a retrospective cohort study of hospitalized adult patients with positive C. difficile NAAT and negative toxin antigen results at University of Iowa Health Care between 9/1/2021 and 9/30/2022. Chart review was conducted to obtain patient age, immunocompromised state, involvement of infectious disease (ID) consultant, prior C. difficile test status and treatment course. Results Of the 295 NAAT-positive C. difficile results, 210 (71%) were toxin antigen negative. Patient characteristics and frequency of common risk factors are shown in Table 1. C. difficile colonization was detected ≥4 days after admission in 111 (53%), with internal medicine and stem cell transplant (SCT)/hematologic malignancies (HM) services having the most patients with colonization (31% and 18%, respectively). Antibiotic use in the prior 30 days was reported in 154 (73%). Of 134 (64%) patients on systemic antibiotics at time of C. difficile testing, antibiotics were discontinued in 29 (22%). Only 43 patients (20%) did not receive any treatment for C. difficile. Median treatment duration was 10 days with 133 (80%) being treated for 10-14 days (Table 2). ID consult was obtained in 48 (23%) and recommended no treatment initially in 11 (23%), 4 of whom were started on treatment in subsequent days. Oral twice a day vancomycin for prophylaxis was given to 28 (13%) patients, 19 (68%) of whom were immunocompromised (16 SCT/HM and 3 solid organ transplant). Conclusion Patients with C. difficile colonization have similar risk factors as patients with CDAD. Understanding treatment patterns for C. difficile colonization can guide educational activities. Most colonized patients received C. difficile treatment and/or prophylaxis, likely due to ongoing systemic antibiotic use and immunosuppression. Benefit of treatment and prophylaxis in colonized patients remains an unresolved issue. Disclosures Kelly M. Percival, PharmD, Gilead Sciences Inc: Advisor/Consultant

PSX-20 Effects of a Phytogenic Supplement Pre- and Post-Calving on Colostrum Yield and Composition, and Cow-Calf Performance

Abstract Objectives were to determine the effects of a phytogenic supplement (a blend of turmeric, capsicum, and pepper extracts), fed pre- and post-calving, on colostrum yield and quality, and cow-calf performance. Twenty-three (BW = 532.08 ± 9.13 kg; age 36 mo) crossbred Angus cows were randomly assigned to 3 treatment groups: 1) Not supplemented (CON, n = 7); 2) supplemented with 250 mg· animal-1· d-1 (PHYT250, n = 8); or 3) supplemented with 500 mg· animal-1· d-1 (n = 8) of the phytogenic supplement. Cows were individually fed the supplement from 30 days (± 5 days) pre-calving (d –30) to 60 days (d 60) post-calving. Total colostrum from the left-rear quarter was collected on d 0 (pre-suckling), and d 1, d 2, and d 3 post-calving. Colostrum weight and volume were recorded, and samples (50 mL) were collected and analyzed for percentages of fat, protein, milk urea nitrogen (MUN), other solids, and lactose. Cow BW was obtained on a biweekly basis from d -30 to d 60. Calf BW was measured on d 0, d 30, d 42, and d 60. Data were analyzed as repeated measures using the MIXED procedure of SAS with treatment, day, and their interaction as fixed effects and animal as a random effect. No treatment by day interaction (P = 0.42) nor main effects of treatment and day (P ≥ 0.25) were observed for colostrum volume. Fat concentration tended to be greater (P = 0.07) for PHYT250 and PHYT500 compared with CON. A treatment × day interaction (P = 0.05) was observed for MUN, with PHYT250 being greater than PHYT500 but similar to CON on d1. There was no difference between treatments for all other days for MUN. No treatment by day interactions (P ≥ 0.15) or main effects of treatment (P ≥ 0.16) were observed for other components. However, there were main effects of day (P ≤ 0.02) with protein being greater on d 0 compared with all other days, and other solids and lactose being greater on day 3 compared with all other days. Main effects of day were observed for cow and calf BW (P ≤ 0.01). Cows were heavier on d 28 and d 42, while calves were heavier on d 56 compared with all other days. Even though we did not observe differences in performance, our results suggest that feeding a phytogenic supplement 30 d pre-calving up to 60 d post-calving may impact fat and MUN in colostrum. To complement these data, we are currently evaluating concentrations of immunoglobulins (IgA, IgG, and IgM) in colostrum and calf blood. Further, milk collections will be performed in specific time-points up to weaning, to evaluate any possible effects of our treatments in milk yield and composition.

Charged Fibrous Dressing to Promote Diabetic Chronic Wound Healing.

Diabetic chronic wounds cause a significant amount of pain to patients because of their low cure rates and high recurrence rates. Traditional approaches to treating diabetic chronic wounds often involve the delivery of drugs or cytokines that regulate the microenvironment and eliminate bacterial infection in the wound area, but they are passive in controlling cell behaviors and may lead to drug resistance. Emerging drug-free wound treatments are important for convenient, effective, and safe treatment strategies. However, the current approaches cannot fully promote tissue regeneration or prevent bacterial infections. Here, the efficacy of a negatively charged fiber dressing in promoting diabetic chronic wound healing is investigated. The negatively charged fiber dressing can generate reactive oxygen species to inhibit bacterial reproduction with the assistance of ultrasound during the inflammatory phase. Furthermore, the dressing provides an electrostatic field that regulates cellular behavior during the inflammatory and proliferative phases. In particular, the dressing can promote fibroblast migration and induce macrophage polarization and neovascularization without any additional drugs. It is demonstrated that this strategy enables the healing of diabetic chronic wounds in a mouse model, achieving effective wound closure over a 12-day treatment cycle and providing a drug-free therapeutic strategy for diabetic chronic wound care.

Surgical Treatment of Postpartureint Omphalitis in Calves

The study was conducted in Al-Muthanna Province-Iraq from January to June 2022, aimed at diagnosing and treating cases of omphalitis in newborn calves. The study included 15 calves infected with umbilical inflammation for 3-30 days after parturition. The animals were divided into three groups according to the severity of the infection and the clinical signs. The study showed that omphalitis in calves occurs in the first days after parturition due to infection of the navel with microorganisms that lead to inflammation and the emergence of clinical signs that include swelling and abscess collection, in addition to high temperature of the animal and lack of appetite. Some cases necessitated surgical intervention to open the swelling and drain the abscess by placing bacteria-killing solutions inside the wound to ensure that the place is sterilized and that the abscess does not collect again. The cases were also treated with antibiotics, antipyretics, and vitamins with repeated treatment for several days until the animal fully recovered. Keywords:- surgical treatment, omphalitis, calves.

Distribution of Opportunistic Pathogens in People Living with HIV at a University Hospital in Istanbul over a One-Year Treatment Period and Its Association with CD4 T Cell Counts.

Among sexually transmitted diseases, HIV causes very serious clinical manifestations that can lead to death. As a result, millions of people have to live with this problem that threatens their health. The virus attacks the immune system of the host, especially CD4+ T lymphocytes, causing the suppression of the immune system. CD4, CD8 counts, and HIV RNA viral loads are monitored in HIV-infected patients with antiretroviral treatment, and CD4 counts play an important role in determining the effectiveness of the treatment. Despite the advances in treatment in the present day, opportunistic infections are the main cause of morbidity and mortality in these patients, and the evaluation of immunological parameters is valuable for the prognosis of the disease in this process. In the present study, the purpose was to investigate the opportunistic infections faced by naive HIV-positive patients who applied to our laboratory and were diagnosed between 2019 and 2022 during their one-year treatment period, and the correlation of the immunological parameters was also evaluated retrospectively using the hospital automation system and laboratory data. A total of 107 opportunistic causative microorganisms were identified in 87 of the 230 HIV-positive patients over one year. T. pallidum was detected in 43 (18.6%) of these patients, Cytomegalovirus (CMV) in 32 (13.9%), Epstein-Barr virus (EBV) in 9 (3.9%), Hepatitis B virus (HBV) in 10 (4.3%), C. albicans in 7 (3%), M. tuberculosis in 3 (1.3%), Hepatitis C virus (HCV) in 2 (0.8%), and C. glabrata in 1 (0.4%) patient. Although mono-agent co-infections were determined in 69 of 87 people living with HIV, two-agent co-infections were detected in 16 HIV patients, and three-agent co-infections were identified in two HIV patients. Considering the correlation between the CD4/CD8 ratio and infection positivity, a moderate negative correlation was determined with HIV RNA viral load and CMV infection. The CD4/CD8 ratio had a low negative correlation with EBV and C. albicans infections. It was also found that the follow-up of HIV RNA load in the diagnosis of T. pallidum, CMV, EBV, and C. albicans may be meaningful. Opportunistic infections mainly affect immunosuppressed patients and can be prevented with effective treatment. Although it is already known that HIV patients may face different infections during their treatment, it was concluded that more attention should be paid to T. pallidum, CMV, EBV, and C. albicans agents. These infections should be routinely monitored with HIV viral load and the CD4/CD8 ratio.