Neubauer Counting Chamber Research Articles

Photoimmunotherapy of Prostate Cancer With Antibody and Fab Fragments Targeting the Prostate Specific Membrane Antigen.

The standard treatment for localized prostate cancer involves surgical removal of the prostate with curative intent. However, when tumor cells persist in the operation site, there is high risk of local recurrence and tumor spread, leading to stressful follow-up treatments, impaired quality of life, and reduced overall survival. This study examined photoimmunotherapy (PIT) as a new treatment option for prostate cancer cells. We generated conjugates consisting of either a humanized antibody or Fab fragments thereof targeting the prostate specific membrane antigen (PSMA), along with our silicon phthalocyanine photosensitizer dye WB692-CB1. PSMA-expressing prostate cancer cells were incubated with the antibody dye or Fab dye conjugates and cell binding was measured using flow cytometry. Cells were irradiated with varying doses of red light for dye activation, and cytotoxicity was determined by erythrosin B staining and subsequent analysis using a Neubauer counting chamber. Specific cytotoxicity was induced with the antibody dye conjugate in the prostate cancer cells in a light dose-dependent manner. Treatment of the cells with the Fab dye conjugate resulted in lower cytotoxicity, which could be attributed to a reduced binding affinity and a reduced dye uptake of the Fab fragment. Our new antibody dye and Fab dye conjugates offer potential for future intraoperative PIT in patients with localized prostate cancer, with the aim to ensure complete removal of tumor cells from the surgical area, to avoid local recurrence, and to improve clinical outcome.

Effect of Giving Jamblang (Syzygium cumini) Leaf Extract on Testosterone Serum Level and Concentration of Spermatozoa in Male Rats Intoxicated with Lead Acetate

Lead is a source of free radicals in the environment. Lead accumulation in the blood generates increasing reactive oxygen species (ROS) in the body and various health problems, including male infertility. One effort that can be made to prevent this problem is to use antioxidants from natural sources such as jamblang (Syzygium cumini) leaf extract. This study is experimental with a randomised posttest-only control group design method. The object of this study was 30 white rats (Rattus norvegicus strain Wistar) which were divided into five groups (K-, K+, P1, P2, and P3). Lead acetate was given as much as 40 mg/kgBW in groups K+, P1, P2, P3 orally for 30 days. Jamblang (Syzygium cumini) leaf extract was given 50 mg/kgBW in the P1 group, 150 mg/kgBW in the P2 group, and 300 mg/kgBW in the P3 group orally for 30 days. Testosterone levels were examined using blood serum taken through the retro-orbital vein and examined using the Enzyme-Linked Immunosorbent Assay (ELISA) method. Spermatozoa were taken from the vas deferens and counted using the Neubauer counting chamber. The average level of testosterone serum was at K-2.47 ± 0.59 nmol/ml, K+ 2.23 ± 0.45 nmol/ml, P1 2.35 ± 0.65 nmol/ml, P2 2.56 ± 0 .73 nmol/ml, P3 1.96 ± 0.59 nmol/ml, there was no significant difference statistically (p > 0.05). The average concentration of spermatozoa at K-32.06 ± 2.66 million/ml, K+ 19.45 ± 8.07 million/ml, P1 27.05 ± 1.89 million/ml, P2 28.65 ± 1.10 million/ml, P3 27.11 ± 5.64 million/ml, statistically there was a significant difference (p < 0.05). There was no effect of giving jamblang (Syzygium cumini) leaf extract on testosterone serum levels and there was an effect of giving jamblang (Syzygium cumini) leaf extract on the concentration of spermatozoa of male Rattus norvegicus rats intoxicated with lead acetate. Keywords: lead acetate, jamblang leaf extract, testosterone, spermatozoa

Development of reference intervals for serum biochemistry and haematology of juvenile green sea turtles (Chelonia mydas) in a Thai rehabilitation centre.

No reference intervals for serum biochemistry and haematology of sea turtles in Thailand exists to assist veterinarians who are responsible for sea turtle health management and treatment. This study determined serum biochemistry and basic haematology of healthy juvenile green sea turtles (n = 92) in captivity in Thailand following the American Society for Veterinary Clinical Pathology (ASVCP), Quality Assurance and Laboratory Standards Committee (QALS) guidelines for the determination of reference intervals in veterinary species. Biochemistry tests, including blood urea nitrogen, creatinine, uric acid, alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase were analysed using an IDEXX VetTest Chemistry Analyzer. Haematology parameters were measured manually using a microhaematocrit for packed cell volume (PCV), Neubauer counting chamber for red blood cell count and cyanmethemoglobin method for haemoglobin concentration. mean corpuscular volume and mean corpuscular haemoglobin concentration were calculated using the PCV, red blood cell count and haemoglobin. Turtles in this study were found to have higher mean values for PCV (28.70%), haemoglobin (92.13 g/L), mean corpuscular haemoglobin concentration (327.03 g/L), uric acid (247.15 μmol/L), alanine aminotransferase (16.53 IU/L), aspartate aminotransferase (209.44 IU/L), and alkaline phosphatase (245.08 IU/L) compared to sea turtles in Brazil. The reference intervals established using high numbers of healthy turtles in this study will assist veterinarians with diagnostic and treatment decisions when evaluating laboratory results for juvenile green sea turtles.

Modification of the Calculation of the Examination of the Platelet Count Using 1 Large Box of Leukocytes in the Improved Neubauer Counting Room

The direct examination of the platelet count using the improved Neubauer counting chamber is carried out by microscopic examination of 25 Medium Boxes of Erythrocytes, this method requires several field shifts so that more accuracy is needed when carrying out the examination. Modification of the calculation of the platelet count by using 1 large box of leukocytes in the improved Neubauer counting chamber requires only 1 field of view at the time of calculation so that it does not take a long time and is more thorough when carrying out microscopic platelet counts. The purpose of this study was to determine the number of platelets using the calculation of 25 squares while the erythrocytes and the number of platelets using a modification of the calculation of 1 large box of leukocytes. This type of research is a quasi-experimental which aims to determine the effect that arises due to certain treatments for examining the platelet count using 25 boxes while the erythrocytes and modification of the calculation using 1 large box of leukocytes. It was carried out at the Hematology Laboratory, Department of Medical Laboratory Technology, Poltekkes Kemenkes Makassar. This research was carried out to start in January-December 2022. Examination of the number of platelets in the counting chamber with 25 Medium Boxes of Erythrocytes and 1 Large Box of Leukocytes, from the 30 data obtained for each method after a different test using the Mann Whitney test obtained a significant p value of 0.234 > 0.05 so that it can be concluded that there is no difference in the number of platelets in 25 Medium Boxes of Erythrocytes and 1 Large Box of Leukocytes. This indicates that examination of the platelet count using 1 large box of leukocytes can be used as a manual examination method

Total Leukocyte Count in Rattus norvegicus after Duffy Binding-Like 2β-Plasmodium falciparum Erythrocyte Membrane Protein 1 Recombinant Protein Injection: The way to a Peptide-based Malaria Vaccine Development

ABSTRACT Introduction: Severe malaria caused by Plasmodium falciparum is mediated by the P. falciparum erythrocyte membrane protein 1 (PfEMP1). It has a DBL2β domain that specifically binds to the intercellular adhesion molecule-1 (ICAM-1) receptor that lies in endothelial cells of many vital organs and is involved in malaria pathogenesis. Antibody against the DBL2β-PfEMP1 protein correlates with a reduced risk of severe malaria, making it a potential malaria vaccine candidate. This study aimed to examine total leukocytes after serial DBL2β-PfEMP1 recombinant protein injection to determine its immunogenicity. Settings and Design: This was an experimental study using pre-post control groups design. Methods: Samples were male rats aged 2-3 months with a weight of 150-350 g. Rats were injected 3x with 100 µg, 150 µg, and 200 µg of the purified DBL2β-PfEMP1 recombinant protein in the three-weeks interval. Blood samples were collected on days 0, 8, 29, and 50, and total leukocytes were counted using the improved Neubauer counting chamber and observed under a microscope. The data were analyzed using the Friedman test, Kruskal Wallis test, and Mann-Whitney test. Results: The lowest leukocyte level was at the pre-injection, and the highest level was after the third injection. There was a significant increase in leukocytes (P < 0.05) in all treatment groups but no increase in the control group. A dose of 100 µg of DBL2β-PfEMP1 recombinant protein showed the best response in inducing the increase of total leukocytes. Conclusion: The DBL2β-PfEMP1 recombinant protein could induce the higher leukocyte level in each injection, especially after the third injection.

New Zealand Rabbit Spermatozoa Concentration after Giving of Beluntas Leaf Tannin Extract (Pluchea indica)

The purpose of this study was to determine the concentration of spermatozoa in New Zealand rabbits after giving of various types of beluntas leaf tannin extract. This type of research is experimental research. The research design is a one-group post-test design. A sample of 16 rabbits. The sampling technique used is purposive random sampling. The research was conducted at the Chemical Engineering Laboratory of the State Polytechnic of Malang, the Laboratory of Kesimma Medica Malang, the Integrated Laboratory, and the Biology Laboratory of the University of Muhammadiyah Malang. The study was carried out from August to September 2019. The study design used a completely randomized design (CRD) with 4 treatments, namely hydrolyzed, condensed, and pure tannin extract of beluntas leaves and the control group was given aquades orally with a dose of 3 ml/kgbb with 4 replications. repetition times. The procedure of this research 1) Preparation of extract of beluntas leaf tannins, 2) Phytochemical test of extract preparations, 3) Treatment of rabbits as experimental animals, 4) Surgery and collection of rabbit spermatozoa, 5) Observation of spermatozoa using a microscope, 6) Calculating the concentration of spermatozoa. The method of data collection was by calculating the concentration of spermatozoa in each treatment using a hemocytometer and a Neubauer counting chamber using a binocular microscope. Data analysis with one-way Anova test and Duncan 5%. The results showed that the dosage of 3ml/kgbb of various types of beluntas leaf tannin extract had an effect on the spermatozoa concentration of New Zealand rabbits (sig<0.05). The findings of this study are that the best treatment that can reduce the number of spermatozoa in New Zealand rabbits is the condensed tannin extract of Beluntas leaves.

Sublethal Effects of Arsenic on Oxygen Consumption, Hematological and Gill Histopathological Indices in Chanos chanos.

Background: The current study was performed aiming to evaluate possible changes in the effect on oxygen consumption, hematology and gill histopathological parameters in fish (Chanos chanos) upon exposure to sublethal concentration of the metalloid arsenic. Methods: Bioassay tests were conducted for determining the LC50 values of arsenic for 96 h. Oxygen consumption in control and arsenic-exposed fish was estimated using Winkler’s method. Red blood corpuscular (RBC) count was examined with a Neubauer counting chamber under a phase contrast microscope. Hemoglobin (Hb) was estimated following the acid hematin method. Histopathological studies were carried by processing and staining the gill tissues with hematoxylin and eosin in accordance with standard histological techniques. They were then subjected to examination under a scanning electron microscope. Results: Chanos chanos exposed to 1/10th of LC50 (24.61%) for a period of 30 days exhibited a maximum decline in the rate of respiration, followed by a decline in RBC and Hb above 45.59% and 51.60%, respectively. Significant toxic lesions encompassing fused gill lamellae, detached gill epithelium, hyperplasia and hypertrophy of respiratory epithelium became heavy handed on the 30th day. Conclusion: Information synthesized from our study serves to be useful in monitoring and managing (As) contamination in the aquatic environment.

The Potency of Anadara nodifera Shell as Natural Testosterone Booster for Male Canary (Seriunus canaria)

Anadara clam shell contains high Zinc. This microelement is recognised as acting natural aromatase blocker (NAB) that capable of boosting testosterone level. The aim of this study was to evaluates the testosterone level of Canaries following Anadara nodifera administration. Adult male Canaries (n = 27) were group into 3 with 9 repetition of each i.e T0 [control, was given by Carboxymethyl Cellulose Sodium (Na-CMC)], T1 (Zinc Sulphate 0.013 mg/30 g BW), and T2 (NAB 0.3 mg/30 g BW). The T2 treatment was NAB or shell powder of Anadara nodifera which was diluted 1:1 in Na-CMC. The birds were treated orally for 21 days. The complete blood cell count was performed in a haemo analyser except the erythrocyte was calculated manually in the Improved Neubauer counting chamber. Testosterone level was measured in ELISA. Subsequently gen CYP19 aromatase in syrinx, brain and testis were identified employing IHC and descriptively analysed. The result indicated significant elevated level of testosterone (p<0.05). The expression of CYP19 aromatase receptor was depressed in syrinx, brain and testis. However there was no significant changes on the blood profile (p>0.05). To conclude, Anadara nodifera at 0.3 mg/30 g BW was capable of acting as NAB to promote testosterone level. This was confirmed by the reduction of CYP19 aromatase gene receptors in syrinx, brain and testis. However NAB treatment was not significantly affecting routine blood profile.

Considerations when using microplates and Neubauer counting chamber in ecotoxicity tests on microplastics

The ubiquity of microplastics raises issues regarding contamination control measures and laboratory practices. The objective was to adapt the use of counting chambers and plastic microplates on the ecotoxicity evaluation of microplastics. Counting chambers, originally used to quantify cells, can also be used to count high concentrations of microplastics (<100 μm) used in laboratory assays. By decontaminating the chamber and mixing the test solution with Nile Red (1:1), fluorescent particles can be easily counted under optical microscopy. Microplate wells, due to their composition, can be contaminated or release microplastics to the test medium, which can interfere with the results of ecotoxicity assays or spectroscopy readings. A cleaning method based on ethanol was developed, which effectively removed particles by 91% without interfering with microalgae yield. Besides providing practical applications that can improve ecotoxicity assays, this work intends to raise awareness on the need to adapt laboratory practices when working with microplastics.

Fermentation of Sweet Sorghum Syrup Under Reduced Pressure for Bioethanol Production

Background:Production of bioethanol from sweet sorghum (Sorghum saccharatum) is a promising “green” energy source that can help to reduce energy dependence on petroleum products, to decrease greenhouse gas emissions, and fight environmental pollution. As an additional benefit, it can promote the exploitation of new uncultivated agricultural lands and favor establishing integrated agro-industrial energy independent enterprises. The alcoholic fermentation under reduced pressure may prevent the accumulation of high ethanol concentrations in the cultured broth and thus may create favorable conditions for the highest productivity of yeastSaccharomyces cerevisiae.Objective:Elaboration of optimal conditions for sweet sorghum syrup fermentation under reduced pressure.Aim:To determine the parameters of sweet sorghum syrup fermentation byS. cerevisiaeunder the conditions of constant and periodic reduced pressure for the highest bioethanol production efficiency.Methods:The sweet sorghum was grown in a temperate continental climate region of Northern Ukraine. The parameters of diluted stem syrup andS. cerevisiaefermentation under reduced and atmospheric pressure were established and controlled by chemical, biochemical and physicochemical methods. The yeast cells were dyed with methylene blue and counted using a microscope and a Neubauer counting chamber. The obtained data have been statistically analyzed.Results:It has been established that a periodic vacuum extraction with short-term heating of the medium to the boiling point is the most promising procedure for bioethanol production. Periodically reduced pressure fermentation of sweet sorghum diluted syrup resulted in 55% higher bioethanol productivity compared to atmospheric pressure fermentation. Such treatment enables to maintain the concentration of ethanol in the medium below 5.5% vol., which does not significantly inhibit the productivity of industrial yeast strains and allows adding a nutrient with the subsequent continuation of the cultivation process. The resulting distillate requires less energy for further dehydration.Conclusion:The sweet sorghum syrup does not contain substances that inhibit yeast cells although nitrogen and phosphorus supplements are required to support efficientS. cerevisiaegrowth. The optimal technology, elaborated in this research, consists of repeated cycles of fermentation under reduced pressure (to the level of vacuum) for boiling the cultured broth. This technology provides the highest bioethanol output, high efficiency, and productivity of the overall process.

Coagulation Parameters among Women with Obstetric Complications in Specialist Hospital Sokoto, Nigeria

Background and Aim: Obstetric complications are part of the leading causes of maternal mortality worldwide. This study was carried out to investigate the effect of complications on the Prothrombin time (PT), activated partial thromboplastin time (APTT), and platelet count (PLC). Women with obstetric complications were recruited as subjects). Pregnant women without complications were included as controls.&#x0D; Study Design: This is a case-control study.&#x0D; Place and Duration of Study: The study took place in Obstetrics and Gynaecology Department of Specialist Hospital Sokoto and the duration was six months.&#x0D; Methodology: Pregnant women aged 18–41 years (mean age 29.5 years) were recruited for the study. The determination of PT and APTT was done by manual methods using commercially prepared Agappe reagent kits, whereas PLC was done by manual methods using a haemocytometer (Improved Neubauer counting chamber). Data were analyzed using SPSS version 23.&#x0D; Results: The results of PT and APTT were significantly higher among women with obstetric complications (14.26±0.23 seconds and 31.32±0.70 second) compared to normal pregnant women (13.00 ± 0.13 seconds and 29.66±0.33) (P&lt;0.05). The PLC was significantly lower among women with obstetric complications compared to women without obstetric complications (P&lt; 0.05).Obstetric complications in subject show a significant association when compared with PT and APTT (P&lt; 0.05). The age group of subjects shows a significant association when compared with APTT (P&lt; 0.05).&#x0D; Conclusion: Findings from this study have shown that obstetric complications causes decrease in platelet count and prolongation in PT and APTT. There is need to assess the PT, APTT and Platelet count routinely for pregnant women to improve the ante-natal care in Specialist Hospital Sokoto.

Absolute eosinophil count as a diagnostic and prognostic marker compared to C- reactive protein and Procalcitonin in patients with sepsis

ObjectiveThis study was undertaken to find out the usefulness of Absolute eosinophil count as a biomarker for sepsis, especially in Indian settings where cost constraints are the rule. MethodsA prospective observational study was carried out over a period of one year in KIMS Al Shifa Super Speciality Hospital which is a tertiary care referral hospital in Malabar region of Kerala. Patients admitted in the ICU with a diagnosis of Systemic Inflammatory Response Syndrome (SIRS) were included except those with hematological cancer, HIV infection, previous eosinophilia due to any cause, trauma patients or those on long term systemic steroids. Absolute eosinophil count estimation at time of admission, and on fifth day, were done manually using Neubauer counting chamber and confirmed using 5 part hematology analyzers ADVIA 2120 (Peroxidase method). The statistical analysis was carried out using the software, Statistical Software for Social Sciences 20.0. ResultsThe mean value of AEC in different categories of sepsis group was found to be significantly low in septic shock and refractory septic shock patients compared to other categories (p = 0.04), whereas CRP levels were significantly higher in all sepsis groups (p < 0.001). When the diagnostic performance of AEC, CRP and PCT were evaluated, Procalcitonin showed maximum sensitivity (89.5%) on day 1 with a cut off value of 0.5 micogm/L in diagnosing sepsis from SIRS patients. AEC shows maximum specificity (95.65%). ConclusionIt is concluded that Absolute eosinophil count below 50 cells/mm3 is a very specific but poorly sensitive indicator to differentiate sepsis from SIRS.A low value of eosinophil in SIRS patients should alert the physician regarding the possibility of severe sepsis and should direct treatment plan accordingly.

Improvement of cell counting method for Neubauer counting chamber.

BackgroundWe compared the cell counting accuracy of the conventional method and the improved method by using Neubauer counting chamber.MethodsIn the improved method, all the border cells were counted and then divided by two; while, in the conventional method, only border cells on the two boundaries (top and left) were counted.ResultsAbout 55.814% of the samples showed more accurate results by improved counting method, about 38.372% had more accurate results by conventional counting method, and about 5.814% were counted with similar counting error by both methods. The improved method had significantly smaller counting error than conventional method (P < .05). The distribution ratio of the border cells was an independent factor for counting accuracy (P < .05).ConclusionTogether, the improved counting method can reduce the counting error of the Neubauer counting chamber to some extent, assess the distributing uniformity of border cells, and help to eliminate the samples with large differences in distribution.

Hematological analysis of Ctenopharyngodon idella, Megalobrama amblycephala and Pelteobagrus fulvidraco: Morphology, ultrastructure, cytochemistry and quantification of peripheral blood cells

The grass carp (Ctenopharyngodon idella), blunt snout bream (Megalobrama amblycephala) and yellow catfish (Pelteobagrus fulvidraco) are economically important fishes in China. Fish hematological features, especially the type and number of peripheral blood cells, are crucial for the evaluation of fish health and the diagnosis of fish diseases. Since the automatic blood cell count equipment for human is not suitable for fishes, the manual method is critical in the quantification of fish blood cells. To make sense of the comparison and interpretation of the blood cell count studies in different articles, the standardization of blood cell classification is necessary. In this study, erythrocytes (red blood cell, RBC), thrombocytes (TC) and leucocytes (i.e. white blood cells, WBC, including lymphocytes, neutrophils and monocytes) were well distinguished in blood smears with Giemsa staining and confirmed by transmission electron microscopy. RBC, TC and WBC were directly counted with an improved Neubauer counting chamber in a modified diluting solution. The differential leucocyte count (DLC) was carried out in blood smears. In view of the labeling characteristics of peroxidase (PO) positivity in neutrophils and non-specific esterase (α-ANAE) positivity in monocytes, PO positive cell percentage and α-ANAE positive cell percentage were also determined in cytochemistry staining smears. No difference was found for the percentages of neutrophils and monocytes between Giemsa staining and cytochemistry staining. The standardized classification, normal count ranges and sizes of the peripheral blood cells by the present systemic studies will provide useful references for monitoring the health status of grass carp, blunt snout bream and yellow catfish.

Analysis of Morphology and Erythrocyte Count in Rat after Electrical Exposure

Electrical burns cause high mortality and morbidity and various complications, although the incidence is only 4% of all burns. Joule heating phenomenon occurs in electric burn patients which cause changes in erythrocyte morphology resulting hypercoagulability that contribute to the formation of thrombus. Changes in erythrocyte morphology may cause tissue oxygenation disorders and decrease in erythrocyte count leading to hypoxia. Until now there have been no reports of changes in erythrocyte morphology and reduction in the number of erythrocytes in burn patients due to electricity. This study aims to observe and analyze morphological and erythrocyte count in rats after electrical exposure of 140 V for 17 seconds. This is a true experimental laboratories study with post-test only control group design. The sample used were 24 rats consisting of one control group (K) and five treatment groups (P). Treatment groups were distinguished by days of blood sampling that is days 0 (P1), 3 (P2), 7 (P3), 10 (P4), and 14 (P5) post electrical exposure of 140 V for 17 seconds. Assessment in erythrocyte morphology was performed using giemsa staining that observed with 1000x magnification while erythrocyte count was performed using improved neubauer counting chamber. One Way Anova showed that there was no significant change in rats erythrocyte count after electrical exposure of 140 V for 17 seconds (p = 0.110). Descriptive analysis showed that there is morphological changes in erythrocyte morphology after electrical exposure of 140 V for 17 seconds, in terms of shape there are more fragmentosit, microsite, tear drops, stomatocytes, and cell target, in terms of size there are more microsite, and in terms of color there are more hypochromic erythrocytes.&#x0D; Keywords: Burn Injury, Erythrocyte Morphology, Erythrocyte Count

Evaluation of Toxicity Effect of Graded Doses of Moringa oleifera Leaf Extract on Blood Indices Using 20 Adult Wistar Rats

Epidemiological studies have indicated that Moringa oleifera leaves are a good source of nutrition and exhibit antioxidant activities. Moringa oleifera is being used for many medicinal and nutritional purposes, since it is widely consumed by all for nutritional and varying medicinal purposes; however there is limited scientific data available regarding excessive consumption of this plant. This study is carried out to evaluate the effect of Moringa oleifera leaf extract on some blood indices. 20 adult wistar rats were used for the study. They were divided into five groups (A, B, C, D & E) of four animals each. Group A served as the control and was orally administered distilled water; the experimental groups B, C, D, & E were orally administered different graded doses of Moringa oleifera leaf extract as follows 2000mg/kg, 3000mg/kg, 5000mg/kg and 7000mg/kg respectively for twenty one days. Twenty four hours after the last administration, blood samples for haematological analysis were collected through cardiac puncture into sterilized glass tubes containing ethylene diamine tetra acetic acid (EDTA) as anticoagulant. Packed cell volume estimation was done using microhaematocrit method. Sample dilution was done using Turks fluid and cell count using improved neubauer counting chamber. Platelet count was estimated with ammonium oxalate and cell count using counting chamber. The result of this study suggests that excessive consumption of aqueous Moringa oleifera leaf extract may have little effect on blood parameters with slight changes in white blood cells.

269 EFFECT OF DIFFERENT COLLECTION FREQUENCY INTERVALS ON THE QUALITY OF SEMEN OF FRENCH BULLDOGS

Dogs produce fewer sperm than other species. Furthermore, for French Bulldogs, anatomical peculiarities, low libido, and increased sensitivity to stress could cause further reductions in sperm count. The objective was to compare effects of semen collection at 24- versus 48-h intervals on semen quality of French Bulldogs. Five purebred French Bulldogs, 19 to 48 months old, were subjected to 5 semen collections (24 h apart). After a 30-day rest, collections were repeated, but the interval between collections was 48 h. Semen was collected (all 3 fractions) by digital manipulation without female stimuli. Volume was measured in a 20-mL syringe. Sperm concentration was determined with a Neubauer counting chamber. Motility and vigor were evaluated with a coverslipped drop of semen on a slide (preheated to 37°C). Motility was expressed as a percentage of motile sperm, and vigor was classified on a scale of 1 to 5. Morphology was evaluated by the panoptic method; 100 cells were counted and results expressed as the percentage of normal or defective cells. Effects of collection interval were analysed using PROC MIXED of SAS (animal as subject and collection as a repeated measure), with collections 2 to 5 compared with collection 1 (using the DIFF option). For collection every 24 h, the third, fourth, and fifth sperm collections were lower than the first collection for volume (10.4 ± 1.1, 8.3 ± 1.1, 5.6 ± 1.1, 3.5 ± 1.1, 2.4 ± 1.1 mL), concentration (437 ± 24, 448 ± 24, 370 ± 24, 322 ± 27, 258 ± 31 ×106 sperm mL–1), vigor (4.6 ± 0.2, 4.2 ± 0.2, 3.6 ± 0.2, 3.7 ± 0.2, 3 ± 0.2), and morphologically normal sperm (82 ± 2.2, 83 ± 2.2, 72 ± 2.2, 68 ± 2.5, 66 ± 2.9%). However, when the interval was increased to 48 h, only the fourth and fifth collections were lower (P &lt; 0.05) than the first for volume (11.8 ± 1.1, 10.2 ± 1.1, 8.8 ± 1.1, 6.6 ± 1.1, 2.5 ± 1.1 mL), concentration (447 ± 24, 410 ± 24, 407 ± 24, 322 ± 24, 241 ± 31 ×106 sperm mL–1), vigor (5 ± 0.2, 4.8 ± 0.2, 4.4 ± 0.2, 4.2 ± 0.2, 4 ± 0.2), and sperm with normal morphology (92 ± 2.2, 90 ± 2.2, 87 ± 2.2, 80 ± 2.2, 81 ± 2.9%). Motility decreased (P &lt; 0.05) following the fourth collection at 24-h intervals and decreased (P &lt; 0.05) after the third collection at 48-h intervals. With a 24-h interval, 4 dogs had &lt;60% motility lower at the fifth collection, whereas only 2 dogs had motility &lt;60% on the fifth collection at 48-h intervals. In conclusion, semen collected at 48-h intervals was of better quality than semen collected at 24-h intervals.

Influence of HMG-CoA Reductase Inhibitors on Leptin-Induced Endothelial Cell Proliferation, Migration, and Capillary-Like Tube Formation

Objective: This study investigated the impact of the hepatic hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) on the leptin-induced human umbilical vein endothelial (HUVE) cell proliferation, migration, and capillary-like tube formation. Methods: The HUVE cells were isolated and cultured, and stimulated with leptin, statins (cerivastatin, fluvastatin, simvastatin), mevalonate, farnesyl pyrophosphate, geranylgeranyl pyrophosphate, or methyl-β-cyclodextrin. The endothelial cell proliferation was assessed using the Neubauer counting chamber. The migration of HUVE cells was examined with the planar migration assay. In vitro capillary sprouting was quantified by measuring the sprout length, number, and cumulative sprout length. Results: The HMG-CoA reductase inhibitors significantly reduced leptin-induced proliferation and migration, which was reversed by mevalonate. Further, the inhibitory effect of the statins on leptin-induced migration was shown to be modulated by the prenylation of farnesyl pyrophosphate and geranylgeranyl pyrophosphate. Although stimulation with a leptin showed no significant effect, a marked increase in capillary-like tube formation was observed with a joint stimulation with HMG-CoA reductase inhibitors. Although statins caused inhibition of proliferation and migration, the same dose of the agents amplified the selective growth of capillary-like tube formation. Membranous cholesterol depletion by methyl-β-cyclodextrin showed a weaker effect compared with statins. Conclusion: Through modulation of prenylation, leptin-induced pro-atherosclerotic events including proliferation and migration were inhibited by HMG-CoA reductase inhibitors.

Relações entre a câmara de Neubauer a espectrofotometria utilizadas para a determinação da concentração espermática de catetos (Pecari tajacu)

Em ejaculados provenientes de 28 catetos, verificou-se a existência de relações entre a concentração espermática determinada por meio da câmara de Neubauer e a tramitância observada por espectrofotometria, utilizando comprimentos de onda variando de 530 a 590nm. Os ejaculados apresentaram uma concentração média de 283,9±30,8x106 espermatozoides mL-1, com variação de 30 a 640x106 espermatozoides mL-1. Os valores para tramitância variaram entre 36,9 a 96,3, nos diferentes comprimentos de onda. Não foram detectadas relações significativas entre os dois métodos (P&gt;0,05). Dessa forma, não se recomenda a espectrofotometria para os procedimentos de rotina na determinação da concentração espermática em catetos.

Influence of pre-implant bone augmentation on diving fitness: An in vitro study

Purpose: Autogenous bone grafts are commonly used for pre-implant bone augmentation or defect coverage in the maxillofacial region. Particulate bone or bone blocks without a vascular pedicle are normally used for this purpose. The objective of this study was to investigate the effects of hyperbaric conditions on the cell activity of augmented bone under pressure changes such as those to which combat divers and mine clearance divers are exposed. Materials and Methods: Systematic studies were performed using an animal model (isogeneic Lewis rats). We collected bone chips (n = 4) from rat femurs with a Safescraper; and obtained isogeneic bone blocks (n = 4) with a bone rongeur. Bone samples were exposed to three simulated 45-min dives to 5.0 bar using oxygen or compressed air. Bone chips and blocks that were not exposed to pressure served as controls. After 9 and 15 days of in vitro cultivation, osteoblast proliferation rates were assessed in a Neubauer counting chamber and the results were statistically analyzed. Results: Irrespective of the atmospheric conditions, bone blocks showed significantly higher proliferation rates (P = 0.05) than bone chips. When exposed to compressed air during simulated dives, both groups showed considerably lower proliferation rates than the control groups. When exposed to oxygen, both groups showed significantly higher proliferation rates than the control groups. Conclusion: Depending on the breathing gas used, changes in atmospheric conditions during simulated dives can lead to a decrease in osteoblast activity. Patients who underwent bone augmentation should therefore be advised not to dive using compressed air until osseointegration is complete.