Average Cell Count Research Articles

Quantifying Competitive Fitness in Yeast with High-Throughput Fluorescence Microscopy Imaging.

Competitive fitness is a fundamental concept in evolutionary biology that captures the ability of organisms to survive, reproduce, and compete for resources in their environment. Competitive fitness is typically assessed in the lab by growing two or more competitors together and measuring the frequency of each at multiple time points. Traditional microbial competitive fitness assays are labor intensive and involve plating on solid medium and counting colonies. Here, we describe a method to quantitatively measure competitive fitness using fluorescence microscopic imaging and machine-learning-enabled image analysis to directly count the number of cells from each competitor in the mixed population. This high-throughput, primarily automated, and efficient process gives accurate and reproducible results for competitive fitness. Here, we describe the entire process, from sample preparation through microscopy to quantification, and provide instructions and scripts for the image analysis, fitness calculations, and sample data visualizations. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Sample preparation Basic Protocol 2: Photographing fluorescing and non-fluorescing cells using an EVOS microscope Basic Protocol 3: Counting fluorescing and non-fluorescing cells with Orbit Image Analysis Basic Protocol 4: Getting the average cell counts per well and changing the file names Basic Protocol 5: Calculating competitive fitness using R.

Morphological characteristics of haemocytes and haemogram of the red king crab

Based on the materials of research work in July-August 2023 on board the R/V ‘Professor Boiko’ in the Barents Sea, the haemolymph parameters of the Kamchatka crab Paralithodes camtschaticus are presented. Haemolymph samples were taken from the heart of 9 freshly caught crabs, then preparations were made immediately in a Goryaev chamber and haemocytes were counted in all large squares of the Goryaev chamber. Agranulocytes, semigranulocytes, granulocytes and transparent cells were noted among the haemolymph cells, which is generally similar to the crustacean haemolymph. The size and morphological characteristics of the cells of the haemocytic series of the Kamchatka crab are similar to those of other invertebrate species such as Fenneropenaeus chinensis, Paranephrops planifrons, Hyas araneus, Homarus americanus, Panulirus interruptus, Loxorhynchus grandi, Astacus astacus, Astacus leptodactylus. The dominant type in haemolymph is agranulocytes, their proportion averaged 42.1±5.4% for the moult stages studied. The proportion of semi-granulocytes and granulocytes is 32.2±3.1 and 22.8±3.9%, respectively, and the proportion of clear cells is 2.9±0.6% of all cellular elements in haemolymph. The percentage of haemocytes in haemolymph at different stages of the larval cycle of male Kamchatka crab differed but not significantly at p > 0.05. The total number of haemocytes ranged widely from 1719 to 11250 units/μL, with an average cell count of 4.2±1.1 ×109 cells/L. Differences in hemolymph BSF at different stages of the larval cycle of male Kamchatka crab were also statistically unreliable (p > 0.05).

Plant Spacing Variations on Cell Number and Agar Content in Gracilaria verrucosa

The primary issue at the Gracilaria verrucosa seaweed cultivator is the low productivity associated with current cultivation methods. Analyzing cell number and agar content provides valuable insights into seaweed cultivation productivity, growth activity, nutrient absorption, and chemical composition. This research aims to examine how variations in planting distance affect the cell number and agar content of Gracilaria verrucosa. The research was conducted at the Maranak Experimental Pond Installation, Brackish Water Aquaculture Fisheries Research Institute, located in Marannu Village, Lau District, Maros Regency. Analysis of cell numbers and agar content was performed at the Fisheries Research Institute Laboratory of the Brackish Water Aquaculture Center from June to July 2024. Utilizing a completely randomized quantitative experimental design with four treatments and three repetitions, the research included: Treatment A (planting distance 20 cm), Treatment B (30 cm), Treatment C (40 cm), and Treatment D (50 cm). The results indicated that planting distance significantly impacts cell number, with the 50 cm distance yielding the highest average cell count of 213.00 ± 9.64. Similarly, planting distance affects agar content, with the 50 cm distance producing the highest agar content of 27.31%. Therefore, a planting distance of 50 cm is identified as the most effective, resulting in both the highest number of cells and agar content.

Predictors of CD4 cell count progression over time among adolescents and young adults transitioning to adult-oriented HIV care in Southern Ethiopia from 2017 to 2021: a retrospective cohort study

BackgroundHuman immunodeficiency virus (HIV), a viral infection impacting CD4 cells, requires an understanding of factors influencing CD4 cell counts to reduce acquired immunodeficiency syndrome(AIDS)-related morbidity and mortality. Despite its importance, no studies have examined predictors of CD4 cell count progression post-transition to adult-oriented HIV care in Ethiopia. This study aimed to identify predictors affecting CD4 cell count trends among adolescents and young adults transitioning to adult-oriented HIV care. Therefore, this study aimed to identify the predictors that affect the trends of CD4 cell count over time among adolescents and young adults who transitioned to adult-oriented HIV care.MethodsA retrospective cohort study was conducted among 206 adolescents and young adults who transitioned to adult-oriented HIV care and had at least two CD4 cell counts in eight selected health facilities in Southern Ethiopia. These facilities were chosen due to their high volume of people living with HIV. A multivariable linear mixed effect model was fitted to identify the predictors of CD4 cell count progression through time. Multi-collinearity between variables was checked using the variance inflation factor. Model comparisons were done using Akaike and Bayesian information criteria. Regression coefficients with 95% confidence intervals (CI) and P-value ≤ 0.05 were used to measure the strength of association and significance level.ResultsThe average CD4 cell count was raised by 6.7 cells/mm3 every six months among the study participants. Transitioning at the age of 18 or older (β = 204.71; 95% CI = 81.86347, 327.5574; P < 0.001), being rural residents (β= -119.776; 95% CI = -215.362, -24.189; P < 0.014), having World Health Organization (WHO) stage III & IV HIV (β= -182.161; 95% CI = -318.475, -45.847; P < 0.009), were the predictors for CD4 cell count progression over time.ConclusionBased on the identified predictors of CD4 cell count progression over time among HIV-positive adolescents and young adults transitioning to adult-oriented HIV care in southern Ethiopia, such as observation time, age at transition, residence, and WHO staging, the study highlights the need for interventions focused on tailored support. Specifically, these interventions should target individuals transitioning with less ART time, those younger than 18, those residing in rural areas, and those with advanced WHO staging, to improve their CD4 cell counts over time.

Utility and performance of cell blocks in cerebrospinal fluid cytology: Experience at two teaching hospitals.

Cytology cell blocks (CBs) are not routinely made for cerebrospinal fluid (CSF) specimens. The goal of this study was to identify when CSF CB preparation improves diagnostic performance. Under institutional review board approval, a retrospective review of CSF cytology cases was conducted at a tertiary university-based hospital and an affiliated county hospital. Patient history, CSF volume, final diagnosis, use of stains, and whether the CB was contributory was determined from the cytopathology report. CSF nucleated cell count data was obtained from the medical record. A total of 69 CSF specimens with CBs from January 2006 to March 2023 were identified from 61 patients. The median CSF volume was 8mL (interquartile range, 4-13mL; range, 1-800mL), with immunohistochemical stains performed on 29 (42%) cases. Per cytology report, CB was contributory in 23 cases (33%), not contributory in 34 cases (49%), and not discussed in 12 cases (17%). The median volume was 8mL for cases in which CB was contributory, not contributory, or not discussed. There was no difference in average nucleated cell counts between cases in which CB was contributory versus not contributory (73.9 vs. 40.0, p=.175). CBs for CSF samples were contributory in a subset (33%) of cases. The authors were unable to identify any specific pre-analytic factors, including specimen volume and average nucleated cell counts, for cases in which CB was contributory. Further evaluation is needed to identify if there are scenarios in which CSF CBs should be routinely prepared.

Quantification of the cellular content of platelet-rich plasma harvested after injection of filgrastim versus pegfilgrastim biosimilars: a prospective, single-center, crossover study

IntroductionHematopoietic stem cells (HSCs) appear in peripheral blood (PB) following mobilization due to external events or pharmaceutical agents like filgrastim and pegfilgrastim. Concentration of these into platelet-rich plasma (PRP) can be used in orthopedics. ObjectivesThis prospective, single-center, controlled laboratory study aimed to compare safety profiles of filgrastim and pegfilgrastim administration, quantifying the cellular content of PRP derived from PB post-administration. We hypothesized comparable rates and severity of adverse events (AEs) among participants, along with similar cellular content in the PRP products. MethodsTen healthy male participants underwent a crossover design with 2 interventions separated by a washout period. PB was collected before and after each intervention, producing PRP for cellular comparison in vitro. AEs were monitored for severity and frequency throughout study. ResultsSignificant AEs for filgrastim included fatigue (36%), myalgia (36%), and back pain (9.1%), compared to pegfilgrastim, which included myalgia (31%), fatigue (23%), and back pain (23%). There were no significant differences between AE occurrences (P = .49) nor relationship to study drug (P > .99). Regarding total nucleated cell count (TNC), statistical significance was found in TNC count (P = .010) and TNC concentration (P = .004). All PRP products expressed high levels (>88%) of hematopoietic progenitor cell/HSC-specific CD45+, CD34+, CD45dim+, and hematopoietic progenitor cell/HSC+ markers. ConclusionsThis study demonstrated that filgrastim and pegfilgrastim administration appear safe. Pegfilgrastim is equivalent to filgrastim for the mobilization of white blood cells and cells expressing hematopoietic cell-surface markers into the PB and consequently, into the derived PRP product. Clinical relevanceThis study presents pegfilgrastim as a single-injection alternative to filgrastim for the mobilization of HSCs into the PB and subsequent PB-derived PRP, which is more convenient and less invasive for patients.

Effect of Anterior Uveitis on Corneal Endothelial Cells

Objective: Investigating the impact of anterior uveitis on changes occurring in corneal endothelial cells. Methods: In this prospective cross-sectional study (cases and controls), the research sample included 43 patients (86 eyes) from the ophthalmic clinic attendees at Tishreen University Hospital in Latakia during the period 2023-2024. Results: The average corneal endothelial cell count was lower in the anterior uveitis group, both granulomatous (2074.16±173.70 cells/mm²) and nongranulomatous (2188.38±165.7 cells/mm²) types, compared to the control group (2696.1±136.60 cells/mm²) with a (p-value &lt;0.01) in all age groups studied. The average percentage of hexagonal cells was lower in the both granulomatous (42.41±2.6%) and non- granulomatous (47.0±2.2%) types, compared to the control group (61.4±2.0%) with a (p-value &lt;0.01) in all age groups studied. The average cell volume change coefficient was higher in the anterior uveitis group, both granulomatous (40.16±2.1) and non-granulomatous (34.70±1.08) types, compared to the control group (29.8±1.36) with a (p-value &lt;0.01) in all age groups studied. Host and disease-related characteristics had no effect on the measured results) p-value=0.184(. ECC was negatively correlated to the duration of uveitis (a=-0.513; p-value&lt;0.001), maximum intraocular pressure during the course of disease (a=-0.472; p-value = 0.0001), and the number of attacks (a=-0.515; p-value&lt;0.001). Conclusion: Corneal endothelial cells are affectd in patients with anterior uveitis, and this negative impact increases with the duration of the disease, the number of attacks, and the presence of elevated intraocular pressure. This negative impact is more pronounced in granulomatous uveitis.

Abstract B008: High expression of the exhaustion markers PD1 and PD-L1 in non-muscle invasive bladder cancer is associated with poor outcome following Bacillus Calmette-Guérin immunotherapy

Abstract Introduction: The recommended treatment for high-risk non-muscle invasive bladder cancer (NMIBC) is intravesical instillations of Bacillus Calmette-Guérin (BCG). However, 40 % experience recurrence within 5 years despite completing BCG treatment. T cell exhaustion has been associated with poor outcome following treatment with BCG. Here we investigated if T cell exhaustion, characterized by protein expression of PD1 and PD-L1 measured in paired samples obtained before and after BCG treatment could further explain BCG response and help predict outcome in patients with NMIBC. Methods: We included 111 patients with NMIBC, from which we had 183 TURB-T samples obtained before and after BCG treatment. Using immunohistochemistry (IHC) staining of sections from tissue microarrays (TMAs) with triplicate core biopsies, we investigated the protein expression of the exhaustion markers PD1 (clone EP239) and PD-L1 (clone 22c3). Data was analyzed using digital pathology software (Visiopharm®), where tissue areas were defined as either carcinoma or stroma based on PanCK staining. Cells were scored as positive when detection of a nucleus was combined with a positive PD1 or PD-L1 stain, which was based on visually defined thresholds to differentiate between positive and unspecific staining. In total, 167 samples from 105 patients were considered suitable for further analysis (cell count &amp;gt; 200 in carcinoma and stromal areas) with 2-3 cores per sample, resulting in a total of 441 tissue cores. The average cell count was 5732 per core (defined by positive nuclei stain). Stratification of samples into high or low expression was based on a median split of the positive cell fraction. Results: When investigating the number of PD1 and PD-L1 positive cells in the tumors (carcinoma and stroma combined), we observed that PD-1 expression significantly increased with tumor stage in pre- (p=0.002) and post-BCG (p=0.006) tumor samples. PD1 expression was also increased in high-grade tumors compared to low-grade tumors in pre-BCG samples (p = 0.001). Furthermore, we observed a significant association between tumors of higher stage and high PD-L1 expression in the pre-BCG samples (p = 0.007). Patients with low expression of PD1 and PD-L1 in the pre-BCG tumor samples had a superior high-grade recurrence-free survival (HG-RFS) compared to patients with high PD1 (p=0.002) and PD-L1 (p=0.021) expression. In addition, low expression of PD-L1 in pre-BCG tumors was correlated with a better progression-free survival compared to the patients with high PD-L1 expression (p = 0.013). Conclusion: Protein expression of PD1 and PD-L1 in pre-BCG tumor samples were correlated to higher stage and grade as well as worse HGFRS, indicating that T cell exhaustion plays an important role in resistance to BCG treatment. Citation Format: Tine G. Andreasen, Trine Strandgaard, Jørgen B. Jensen, Lars Dyrskjøt. High expression of the exhaustion markers PD1 and PD-L1 in non-muscle invasive bladder cancer is associated with poor outcome following Bacillus Calmette-Guérin immunotherapy [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr B008.

Corneal Endothelial Cells and Central Corneal Thickness in Pseudoexfoliation Patients

Objective: Investigating the impact of pseudoexfoliation syndrome on changes occurring in corneal endothelial cells and central corneal thickness, while assessing the influence of the degree of pseudoexfoliation on endothelial cell changes. Methods: In this analytic case-control study (Cross Sectional), the research sample included 60 patients (120 eyes) from the ophthalmic clinic attendees at Tishreen University Hospital in Latakia during the period 2023-2024. Results: The average corneal endothelial cell count was lower in the pseudoexfoliation group (2218.91±311.5 cells/mm²) compared to the control group (2559.82±290.7 cells/mm²) with a (p-value of 0.0001). The average percentage of hexagonal cells was lower in the pseudoexfoliation group (48.55±14.9 %) compared to the control group (62.32±9.7%) with a (p-value of 0.0001). No statistically significant differences were observed between the research groups in terms of average cell volume change coefficient with a (p-value of 0.9). The average cell area was higher in the pseudoexfoliation group (487.92±71.3 µm²) compared to the control group (419.94±72.2 µm²) with a (p-value of 0.003). The average central corneal thickness was lower in the pseudoexfoliation group (515.93±41.9 µm) compared to the control group (544.22±26.6 µm) with a (p-value of 0.04). We observed increased changes in corneal endothelial cells and central corneal thickness with an increase in the degree of pseudoexfoliation. Conclusion: Corneal endothelial cells and central corneal thickness are affected in patients with pseudoexfoliation syndrome, and these changes increase with severity of pseudoexfoliation.

Breast Implant Illness May Be Rooted in Mast Cell Activation: A Case-Controlled Retrospective Analysis.

To investigate the possible association between breast implant illness (BII) and mast cell activation syndrome (MCAS), which often manifests increased mast cells (MCs) in assorted tissues and may explain BII symptoms. Mechanisms by which implants cause BII symptoms remain unclear, but BII and MCAS symptom profiles heavily overlap, warranting investigation of potential linkage. We retrospectively analyzed 20 implant patients who underwent explantation and total capsulectomy; 15 self-reported preoperatively they had BII (subject group); 5 felt they did not [control group 1 (CG1)]. Five prophylactic mastectomy patients constituted control group 2 (CG2). Subjects and CG1 patients completed BII symptom questionnaires preoperatively and multiple points postoperatively. With CD117 staining, average and maximum mast cell counts (MCCs) in resected tissues were determined. Mean BII symptom score 2 weeks postexplantation was reduced by 77% (P < 0.0001), and 85% by 9 months. Analysis suggested BII in CG1 patients, too, who improved similarly. Among CG2 patients, healthy breast tissue showed mean and maximum MCCs of 5.0/hpf and 6.9/hpf. Mean and maximum MCCs in capsules in BII patients were 11.7/hpf and 16.3/hpf, and 7.6/hpf and 13.3/hpf in CG1 patients. All intergroup comparisons were significantly different (P < 0.0001). MCCs in peri-implant capsules in BII patients are increased; some implanted patients appear to have unrecognized BII. Given that neoantigenic/xenobiotic exposures commonly trigger dysfunctional MCs in MCAS to heighten aberrant mediator expression driving inflammatory and other issues, further investigation of whether BII represents an implant-driven escalation of preexisting MCAS and whether an MCAS diagnosis flags risk for BII seems warranted.

Development and validation of a diagnostic model to differentiate spinal tuberculosis from pyogenic spondylitis by combining multiple machine learning algorithms.

This study focused on the development and validation of a diagnostic model to differentiate between spinal tuberculosis (STB) and pyogenic spondylitis (PS). We analyzed a total of 387 confirmed cases, out of which 241 were diagnosed with STB and 146 were diagnosed with PS. These cases were randomly divided into a training group (n = 271) and a validation group (n = 116). Within the training group, four machine learning (ML) algorithms (least absolute shrinkage and selection operator [LASSO], logistic regression analysis, random forest, and support vector machine recursive feature elimination [SVM-RFE]) were employed to identify distinctive variables. These specific variables were then utilized to construct a diagnostic model. The model's performance was subsequently assessed using the receiver operating characteristic (ROC) curves and the calibration curves. Finally, internal validation of the model was undertaken in the validation group. Our findings indicate that PS patients had an average platelet-to-neutrophil ratio (PNR) of 277.86, which was significantly higher than the STB patients' average of 69.88. The average age of PS patients was 54.71 years, older than the 48 years recorded for STB patients. Notably, the neutrophil-to-lymphocyte ratio (NLR) was higher in PS patients at 6.15, compared to the 3.46 NLR in STB patients. Additionally, the platelet volume distribution width (PDW) in PS patients was 0.2, compared to 0.15 in STB patients. Conversely, the mean platelet volume (MPV) was lower in PS patients at an average of 4.41, whereas STB patients averaged 8.31. Hemoglobin (HGB) levels were lower in PS patients at an average of 113.31 compared to STB patients' average of 121.64. Furthermore, the average red blood cell (RBC) count was 4.26 in PS patients, which was less than the 4.58 average observed in STB patients. After evaluation, seven key factors were identified using the four ML algorithms, forming the basis of our diagnostic model. The training and validation groups yielded area under the curve (AUC) values of 0.841 and 0.83, respectively. The calibration curves demonstrated a high alignment between the nomogram-predicted values and the actual measurements. The decision curve indicated optimal model performance with a threshold set between 2% and 88%. In conclusion, our model offers healthcare practitioners a reliable tool to efficiently and precisely differentiate between STB and PS, thereby facilitating swift and accurate diagnoses.

In vitro effects of uncarboxylated osteocalcin on buffalo Leydig cell steroidogenesis.

Uncarboxylated osteocalcin (UcOCN), a bone derived circulating protein, has been demonstrated to influence steroidogenesis in testicular Leydig cells of murine and human species. However, the role of UcOCN in testosterone biosynthesis remains unexplored in domestic animals. The present study aimed to investigate the impact of UcOCN on the expressions of steroidogenic genes (HSD3β1, HSD3β6, CYP17A1, CYP11A1), testosterone production and GPRC6A receptor localization in buffalo Leydig cells. Leydig cells from the testes of adult Murrah buffalo were isolated, with an average cell count and viability after digestion and Percoll enrichment of 1.43 × 106 cells/g of testes and 78.5%, respectively. Immunophenotyping of Percoll-enriched cell suspension by flow cytometry showed populations of Leydig cells ranging between 69 and 73.9%. Immunostaining confirmed the presence of GPRC6A receptors and CYP11A1 positive Leydig cells. When these cells were cultured and incubated with varying levels of UcOCN (6, 12, 24, and 48 ng/ml) and LH, there was a significant (P < 0.01) increase in testosterone production and up-regulation (P < 0.05) of CYP11A1, CYP17A1, HSD3β1 and HSD3β6 gene expression. In summary, the present study underscored the effects of UcOCN on testosterone biosynthesis, expression of crucial steroidogenic genes and interaction with GPRC6A receptors in buffalo Leydig cells, emphasizing its potential implications in andrology.

A single intranasal dose of essential oil spray confers modulation of the nasopharyngeal microbiota and short-term inhibition of Mannheimia in feedlot cattle: a pilot study

Five essential oils (EOs) were previously characterized in vitro and identified as candidate EOs for the development of an intranasal EO spray to mitigate bovine respiratory disease (BRD) pathogens. In the present study, these EOs were evaluated for their potential to (i) reduce BRD pathogens, (ii) modulate nasopharyngeal microbiota, and (iii) influence animal performance, feeding behavior and immune response when a single dose administered intranasally to feedlot cattle. Forty beef steer calves (7–8 months old, Initial body weight = 284 ± 5 kg [SE]) received either an intranasal EO spray (ajowan, thyme, fennel, cinnamon leaf, and citronella) or PBS (Control; n = 20/group) on day 0. Deep nasopharyngeal swabs were collected on days (d) -1, 1, 2, 7, 14, 28, and 42 and processed for 16S rRNA gene sequencing, qPCR, and culturing. Significant effects of EO on community structure (d1), microbial richness and diversity, relative abundance of some dominant phyla (d1, d2, and d14), and the overall interaction network structure of the nasopharyngeal microbiota were detected. The relative abundance of Mannheimia was lower in the EO calves (4.34%) than in Control calves (10.4%) on d2, and M. haemolytica prevalence on d7 as compared to control calves. Feed intake, average daily gain, feeding behavior, and blood cell counts were not affected by EO treatment. Overall, a single intranasal dose of EO spray resulted in moderate modulation of nasopharyngeal microbiota and short-term inhibition of Mannheimia while not influencing animal performance, feeding behavior or immune response. Our study, for the first time, shows the potential use of intranasal EO to mitigate BRD in feedlot cattle.

NUMERICAL INVESTIGATION ON PULSE DETONATION ENGINE PERFORMANCE UNDER DIFFERENT EQUIVALENCE RATIO

Detonation-based engines have a higher thermal efficiency than deflagration-based engines and are therefore widely studied. In this study, the effect of initiation conditions on the detonation wave propagation and the performance of a pulse detonation engine, one type of detonation-based engine, was numerically investigated. Hydrogen-oxygen mixtures with equivalence ratios of φ=0.8, 1.0, and 1.2 were defined as initiation conditions with constant pressure and temperature. The numerical simulations were conducted using the transient explicit density-based solver in the ANSYS Fluent commercial software. The adaptability of the adaptive mesh refinement method in detonation-based engines was explored in the numerical studies, reducing the average total cell count by a factor of 2.617, and obtaining consistent results according to validation studies. The adaptive mesh refinement method was also used in numerical simulations where different equivalence ratios were defined. It was determined that an increase in the equivalence ratio resulted in an increase in the detonation wave velocity. Also, an increase in thrust distribution at the nozzle exit was observed before the blowdown stage, and the calculated thrust values for φ=0.8, φ=1.0, and φ=1.2 were 248.28 N, 264.5 N, and 270.83 N, respectively.

Outcome of Optical Keratoplasty for Corneal Scar due to Infective Keratitis.

Corneal opacity is an important cause of blindness in developing countries. This study analyzes optical keratoplasty performed for corneal opacity due to infective keratitis. This is a retrospective study of all consecutive cases of optical keratoplasty performed between 2011 and 2014 (four-year period) for healed infective keratitis. Cases with less than two months' followup were excluded during outcome evaluation. Comparison was made between keratoplasty for Microbial and Viral (herpetic) Scar. Ninety-three eyes of 93 patients were enrolled. Fifty-nine (63.4%) were male. Average age of patients was 38.9±19.5 years. Average donor endothelial cell count was 2713±434.5 cells/mm2. Fifty-four (58%) corneal scars were due to microbial keratitis and others were herpetic. Eighty-five (91.4%) had undergone penetrating keratoplasty. Eighty-eight (94.6%) cases were included for outcome analysis. Average follow-up duration was 37±27.5 months. Fifty-two (59%) had clear graft at their last visit. Twenty-three (26.1%) grafts had endothelial failure and 13 (14.7%) grafts failed due to late onset keratitis. Twenty-five (28.4%) had vision of ≥6/18. Rejection occurred in 24(27.2%) and glaucoma in 11(12.5%). Post-operatively viral keratitis in the graft occurred significantly more inViral Scar Group (38.6%, n=15) than in Microbial Scar Group (5.5%, n=3). But there was no significant difference in graft clarity, rejection, vision and secondary glaucoma between the two Groups. Outcome of keratoplasty for post-infectious scars was found fairly satisfactory. Although occurrence of viral keratitis was higher in case of keratoplasty done for Viral Scars, the final result was similar to that of microbial scar.

Overview of Monocyte to Hdl Cholesterol Ratio (MHR) in Patients With Coronary Heart Disease (CHD)

Background: Coronary Heart Disease (CHD) is a disease caused by plaque that accumulates in the coronary arteries, causing narrowing or blockage of blood flow that supplies oxygen (O2) to the heart muscle. Monocytes accumulate in myocardial tissue experiencing hypoxia and play a role in the formation of atherosclerotic plaque. On the other hand, High-Density Lipoprotein (HDL) is known to reduce the accumulation of macrophages and prevent oxidized cholesterol from entering the artery walls. In the process of healing CHD, the number of monocytes decreases and HDL-cholesterol increases. This study aims to show the potential of the monocyte to HDL Cholesterol ratio as a marker for the prognosis of CHD patients.&#x0D; Method: This study used a case-control research design with a research sample of 50 CHD patients and 50 MCU patients at Bhayangkara Hospital, Jambi City.&#x0D; Result: From the results of the examination, the average monocyte cell count was 461.5 cells/μL, HDL-Cholesterol= 48.1 mg/dL and MHR value= 9.6. Meanwhile, in MCU patients, monocytes were 398 cells/μL, HDL-Cholesterol= 50.4 mg/dL and MHR value= 8.1.&#x0D; Conclusion: The MHR value of CHD patients was significantly higher than that of MCU patients. MHR appears to have good potential in the prognosis of CHD patients.

Standardized Quantification of Mast Cells in the Gastrointestinal Tract in Adults

Current data on the normal quantity of mast cells throughout the adult gastrointestinal tract are limited in several domains. These include microanatomic localization of mast cells, standardization of staining and counting methods, and reporting of microscope field of view. To address this lack of reliable reference ranges to facilitate the study of and diagnosis of emerging mast cell-mediated diseases. We examined biopsies obtained from the esophagus, stomach, duodenum, and colon from an unselected cohort. Mean and peak mast cell density were determined on slides stained for tryptase and CD117, and were expressed per high power field (hpf) and surface area (mm2), thus deriving reference ranges (average ± 2 SDs). For the most common hpf surface area (0.238 mm2), upper limits of the derived reference ranges for average/peak mast cells were 0.15/3.67 (esophagus, tryptase), 0.70/5.98 (esophagus, CD117), 22.56/35.30 (stomach, tryptase), 31.32/53.10 (stomach, CD117), 30.28/49.77 (duodenal crypts, tryptase), 41.96/65.26 (duodenal crypts, CD117), 4.98/11.56 (duodenal villi, tryptase), 8.38/14.17 (duodenal villi, CD117), 26.58/41.08 (colon, tryptase), and 35.57/57.92 (colon, CD117). Interobserver variability was moderate to good. There was significant correlation between average and peak mast cell counts. These data help standardize mast cell reference ranges throughout the gastrointestinal tract in adults, which can be used to determine whether abnormal levels of mast cells are present in patients with suspected mast cell-mediated disease. Our data show that the commonly used cutoff of 20 mast cells per hpf irrespective of the gastrointestinal tract segment is an underestimate of an appropriate cutoff in stomach, duodenum (crypt area), and colon.

ANALISIS KETAHANAN IKAN LELE DUMBO (CLARIAS GARIEPENUS) YANG DIINFEKSI BAKTERI AEROMONAS HYDROPHILA DENGAN KONSENTRASI BERBEDA

The purpose of this study was to determine the susceptibility of catfish to Aeromonas hydrophila bacteria, and to know the blood picture of catfish infected with Aeromonas hydrophila bacteria. The method used in this study used an experimental method with a Completely Randomized Design (CRD) or Competely Randomized Design (CRD) (Gomez and Gomez, 1984) with 4 (four) treatments and one control each with 3 replications. The results showed that catfish had a strong susceptibility or resistance to Aeromonas hydrophila bacteria. The Lethal Dose value of 50 was 106 cfu/ml, meaning that Aeromonas hydrophila bacteria was able to kill 50% of the catfish population when it was 5.4 x106 cfu/ml. The lowest average red blood cell count was in treatment A, which was 23.89x106 cells/mm3 and the highest in treatment D, which was around 37.5x106 cells/mm3, while the highest and lowest mean total leukocyte counts were 43.07-82.72x103 cells/mm3, still within the range of values. normal. The average hemoglobin concentration was lower than the normal value range in treatment A, which was 6.52 gr% and in treatment D, it was around 12.42 gr%

Dexamethasone's effect on endometrial inflammatory response in jennies after artificial insemination with frozen donkey semen

Artificial insemination (AI) with frozen donkey semen leads to low fertility rates in jennies. This might be due to an exacerbation of the acute inflammatory endometrial response to frozen semen, as seen in mares with persistent post breeding endometritis. Higher pregnancy rates were obtained in these mares when treated with dexamethasone at breeding time. The aim of this study was to evaluate the effect of dexamethasone on endometrial inflammatory responses in jennies after AI with frozen donkey semen. This study was performed from August to December 2022 in Argentina. Six healthy jennies of proven fertility were included in a crossover experimental design. All animals passed through an estrous cycle with no treatment before crossover between treatments. Jennies were subjected to 1) no AI and no treatment in estrus (control, C); 2) AI with no treatment (D-); and 3) AI with administration of dexamethasone, 50 mg IV at AI time (D+). Post ovulation. deep AI was performed with frozen donkey semen (700 × 106 progressively motile sperm (PMS). Endometrial samples (cytology and biopsies) were taken at estrus and 24 hours post AI. Cytology smears were stained with Diff-Quick™; 200 cells were counted under an optical microscope (Nikon Eclipse™ NI-U) and the percentage (%) of polymorphonuclear neutrophils (PMN) and eosinophils was calculated.Endometrial biopsies were taken from the bifurcation of the uterine body, fixed in 10% buffered formalin prior to hematoxylin-eosin staining. Five fields were selected per sample at 400x and average cell counts were calculated. The effect of treatments on % PMN and % eosinophils in cytology and biopsies were evaluated by the Kruskal Wallis test followed by the Dunn test for multiple comparisons. Values are expressed as median and interquartile ranges. The cytology smears of all animals showed an inflammatory response to semen deposition. There were differences (P = 0.003) between % PMN in C [0.625 (0.50 - 0.94)] and the other two groups [D-: 35.00 (22.50 - 46.00); D+: 39.50 (28.63 - 40.25)]. No eosinophils were observed. The inflammatory pattern in biopsies showed a diffuse infiltration of PMN and eosinophils in the luminal epithelium, stratum compactum and stratum spongiosum. There were differences (P = 0.003) between % PMN in C [0.30 (0.05 - 0.40) and the other two groups [D-: 41.90 (28.60 - 43.50); D+: 28.70 (10.35 - 49.00)]. Eosinophils were different (P = 0.005) for the three groups [C: 0.40 (0.20 - 1.20); D-: 21.50 (11.35 - 60.60); D+: 4.60 (1.30 - 7.75)]. In conclusion, a single dose of dexamethasone at AI in jennies did not decrease endometrial PMN but decreased eosinophils 24 hours after insemination.

Evaluation of Mast Cell Density in Mucoepidermoid Carcinoma and Pleomorphic Adenoma.

Mast cells are round to elliptical cells that originate from bone marrow stem cells and enter the peripheral blood. By releasing inflammatory mediators, these cells are involved in type I hypersensitivity, wound healing, defense against pathogens, increased blood vessel formation, and destruction of the extracellular matrix. There are contradictory results regarding the role of mast cells in tumor lesions. Considering the contradictory results and few studies on the density of mast cells in salivary tumors, the present study investigated and compared the density of mast cells in two common salivary gland tumors. In the cross-sectional study after reviewing the records of patients referred to the Pathology Department of the School of Dentistry and Shahid Sadoughi Hospital in Yazd, 15 blocks of each of the mucoepidermoid carcinoma and pleomorphic adenoma tumors were taken. After Giemsa staining of the samples, the average of stained cells in 10 random fields under 400× magnification was counted. The results were analyzed using statistical tests of t-test, ANOVA, Chi-square, and Mann-Whitney in SPSS ver. 22. The average mast cell counts in pleomorphic adenoma (4.2) was higher than muco-epidermoid carcinoma (1.7) but there was no significant relationship (p= 0.305). In mucoepidermoid carcinoma, the numbers of mast cells increased with increasing tumor grade (low: 0/467 moderate: 1/567 high: 2/983) and there was a significant relationship (p= 0.009). According to the results of the present study, it seems that the mast cells accumulation may be secondarily associated with inflammatory responses due to cell accumulation and tissue destruction by tumor cells.