Hemocytometer Research Articles

Аntiparasitic complex of Ivermectin for treatment of herd horses at gastrointestinal Nematodosis

The purpose of the research is evaluating the efficacy of the antiparasitic complex based on ivermectin obtained by mechanochemical technology against gastrointestinal nematodosis of herd horses.Materials and methods. The efficacy of the ivermectin-based antiparasitic complex was tested in December-February 2019 in the Southern Federal District of the Chechen Republic in a private farm on 22 stabled foals aged over one year. To diagnose gastrointestinal parascaridosis and strongylatosis, feces of foals were examined using the Fulleborn’s method. Helminth eggs in 1 g of feces were counted in a counting chamber of the All-Russian Scientific Research Institute of Helminthology. In order to diagnose oxyurosis, pieces of transparent adhesive tape with prints from perianal folds were transferred to a glass slide and examined for Oxyuris spp. eggs. The antiparasitic complex of ivermectin 2% was administered in a mixture with food at a dose of 0.1 mg/kg by the drug (0.2 mg/kg by the active substance). After a preliminary test of the ivermectin complex efficacy, the therapeutic dose of the drug was titrated on 46 adult horses (aged 2–4 years) from the same private entrepreneur. The horses spontaneously infected with Strongylata, Parascarids and Oxyurises were divided into 5 groups according to the analogue principle. For animals of groups 1, 2, and 3, the drug was administered at a doses of 0.1, 0.15 and 0.2 mg/kg by the AS respectively. The horses of groups 4 and 5 were given the substance of ivermectin at a doses of 0.1 and 0.2 mg/kg by the AS, mixed with combined feed individually. The efficacy of the supramolecular complex of ivermectin was recorded according to the data of coproovoscopic studies using the critical test method according to the Guidelines approved by the World Association for the Advancement of Veterinary Parasitology (1995). Results and discussion. The rate of infection of the foals with Strongylata, Parascarids and Oxyurises was 100, 40 and 55% respectively as resulted from the coproovoscopic studies. The dissections showed that the intensity of infection with Strongylata was equal to 477.4±23.87 individuals per animal, Parascarids 447.1±55.89, and Oxyurises 22.2±2.02. The 100 % efficacy was determined for the antiparasitic complex of ivermectin at a dose of 0.1 mg/kg by the drug (0.2 mg/kg by the AS) against the main gastrointestinal nematodes. The substance of ivermectin at a dose of 0.2 mg/kg showed 87.5% efficacy against Strongylata and Parascarids, and at a dose of 0.1 mg/kg, it showed a lack of efficacy (50.0 and 25.0% respectively). We recognized the dose of 0.15 mg/kg by the AS of the supramolecular complex of ivermectin with one-time individual use in a mixture with food as therapeutic.

Standardisation of lymphatic filariasis microfilaraemia prevalence estimates based on different diagnostic methods: a systematic review and meta-analysis

BackgroundLymphatic filariasis (LF) infection is generally diagnosed through parasitological identification of microfilariae (mf) in the blood. Although historically the most commonly used technique for counting mf is the thick blood smear based on 20 µl blood (TBS20), various other techniques and blood volumes have been applied. It is therefore a challenge to compare mf prevalence estimates from different LF-survey data. Our objective was to standardise microfilaraemia (mf) prevalence estimates to TBS20 as the reference diagnostic technique.MethodsWe first performed a systematic review to identify studies reporting on comparative mf prevalence data as measured by more than one diagnostic test, including TBS20, on the same study population. Associations between mf prevalences based on different diagnostic techniques were quantified in terms of odds ratios (OR, with TBS20 blood as reference), using a meta-regression model.ResultsWe identified 606 articles matching our search strategy and included 14 in our analyses. The OR of the mf prevalences as measured by the more sensitive counting chamber technique (≥ 50 µl blood) was 2.90 (95% confidence interval (CI): 1.60–5.28). For membrane filtration (1 ml blood) the OR was 2.39 (95% CI: 1.62–3.53), Knott’s technique it was 1.54 (95% CI: 0.72–3.29), and for TBS in ≥ 40 µl blood it was 1.37 (95% CI: 0.81–2.30).ConclusionsWe provided transformation factors to standardise mf prevalence estimates as detected by different diagnostic techniques to mf prevalence estimates as measured by TBS20. This will facilitate the use and comparison of more datasets in meta-analyses and geographic mapping initiatives across countries and over time.

Pollen Production in Theobroma cacao L. Genotypes National Type and CCN-51 and Its Relationship with Climatic Factors on the Ecuadorian Coast

Cocoa is a primary source of economic income in the agricultural and agroindustrial sectors of Ecuador. However, cocoa crops are vulnerable to meteorological conditions that affect reproduction processes, including pollen production, which has encouraged the study of factors related to low crop yields and practical implications in breeding programs. The objective of this study was to assess the pollen production per flower in 11 genotypes of <em>Theobroma cacao</em> national type and CCN-51 clone. The number of pollen grains per flower was estimated by collecting 10 flower buds from each genotype for 12 consecutive months and directly counting pollen grains using an optical microscope and a Neubauer chamber. An average of 32,611 pollen grains per flower was estimated, with the largest amount of pollen (44,000) produced by the CCN-51 genotype. There was a moderate positive correlation between the environmental variables and pollen production (temperature, <em>r</em> = 0.33; <em>p</em> = 0.028; precipitation, <em>r</em> = 0.43; <em>p</em> = 0.0025). Since pollen production is an important indicator used for the selection of promising genotypes, the genotypes identified with the highest pollen production should be used in breeding programs.

Seasonal variations of zooplankton diversity in fresh water reservoir of West Bengal, India

Background/Objective: Present study was carried out to identify zooplankton density at Baburpukur pond along with physicochemical properties. In India. Planktonic richness reflects the biodiversity stock. The present study assumes greater importance for biodiversity conservation/ pollution indicator and aquaculture of fish and prawns. Methods/Statistical analysis: Statistical analysis in this experiment was performed by Student\'s t-test. In this test, 0.05 probability, degree of freedom, critical t-value, and calculated t-values were recorded. Here zooplankton number along with physico-chemical parameters was recorded. From these t-values, significant seasonal variations were found in the respective water body. The zooplankton density and physiochemical parameters were recorded during the period of Jun 2018 to February 2020. Diversity of zooplankton has been counted using Sedgwick-Rafter counting chamber. Findings: Four different species of zooplankton were studied such as Daphnia, Cyclops, Cypris and Brachionus. Zooplankton community structure generally changes with temperature, pH of water, free CO2 level, dissolved O2. This study also reveals that zooplanktons have their own peak periods of density which is influenced by the above environmental conditions. Application: At present, this water reservoir is suitable for fish culturing. So several management practices are necessary to conserve this zooplankton density for proper healthy situation of water body. This study is also helpful in understanding the zooplankton diversity of Baburpukur Pond with proper maintaining of aquaculture. Keywords: Zooplankton; biodiversity conservation; pollution indicator; aquaculture

Bone Marrow Aspirate Concentrate (BMAC) for the treatment of 27 patients with peripheral arterial disease grade III & IV. A pilot non-randomized study

Background & Aim Peripheral arterial diseases (PAD), as a member of arteriosclerosis, mostly occur in lower extremity arteries. The morbidity of PAD generally ranged from 3 to 10% and can reach above 15% and it upregulates with aging. Autologous bone marrow aspirate concentrated cells (BMAC) have been shown to be safe and effective for treatment of patients with PAD. Angiogenesis can also be induced by growth factors synthesized by them. Methods, Results & Conclusion A descriptive, observational and pilot study, non randomize, a total of 27 patients with critical non-revascularized limb isquemia grade III and IV (Fontaine) were enrolled and studied between 2015 and 2019. Risk factors like diabetes, hypertension, nicotine, dislipemia were compared. A total up to 120 ml of BM cells was aspirated from left illiac crest, into 0.1 ml heparin treated in 20 ml syringes and collected in Harvest Device Technology. Centrifugation was done under protocol Harvest Tech. 20 ml of the bottom centrifugation with total nucleated cells were obtained. The medium dose obtained in all patients was 897.000.000 of TNC's counted by Neubauer chamber with trypan blue (98% of viability). 40-point sites in the leg was injected by BMAC via intramuscular needle 22G from the last site where the superficial femoral artery is permeable. The procedure starts in the proximal part by echo guided images, in order to locate the area of artery that is still permeable, then injections of 1ml with 22G needle are made in an interval of 1 cm. After one year of follow up of 27 patients (100%), 14,8% (4) have amputation of the critical limb affected, 14,8% have strong relationship with cigarettes (74.1%) and have a Fontaine score 4. The 85,2 % of the patients have a strong improvement in their quality of life, ABI and conserve their leg. An angiography was done pre/post (3 months) intramuscular injection were arteries and neoangiogenic blood vessels could be shown by images. The following conclusions were drawn: 1) intramuscular implantation of BMAC in patients with severe PAD is an effective approach that is associated with short-term induction of neovascularization; 2) clinical improvement in the lower limb, which included increased ABI values, increased leg temperature, and other local signs that are indirect indicators of collateral vessel formation; 3) BMAC is a safe procedure, with no adverse events resulting from cell therapy as a point of care-same day procedure in patients with Fontaine Score III-IV.

Production of Liquid Bio-Fertilizer from Old Coconut Water and Molasses using Consortium Microbes

Increasing issue of returning to nature, the presence of organic fertilizer is one of the things required in agriculture sector. The presence of the liquid organic fertilizer, it is expected to have a positive impact on plants and soil. Moreover, in the manufacturing process, it was utilizing wastes. The liquid fertilizer was made by mixing waste from micro-businesses that produce a porridge (old coconut water) and side product of the sugar factory (molasses). Seven litter solutions from the liquid were mixed using a bioreactor equipped with impeller for 8 incubation days. The isolated microbes were inserted with initial number of 1012 using counting chamber method. As a phosphate source, the phosphate rock was implemented into reactor. Every 4 days, the samples were taken and analysed for knowing the concentration of NPK and C components. Three consortium microbe variables were carried out for making the fertilizer where the source of microorganisms from enrichment cultures + old coconut water + molasses (EOM); soil + old coconut water + molasses (SOM); and only old coconut water + molasses (OM). The highest (%) of N, P, K and C components was detected at 0.09; 0.04; 10.5 and 1.74, respectively achieved by the EOM after 8 day of incubation period. The bio-fertilizer was applied into the paddy test plants with a hydroponic system.

Effect of lead on proliferation, oxidative stress and genotoxic damage of 3T3-L1 fibroblasts.

Lead (Pb) is an environmental and industrial contaminant that still represents a public health problem. In this paper, we investigated the effect of Pb on proliferation, lipid peroxidation and the number of micronucleated cells in exponentially growing 3T3-L1 fibroblasts, a cell line previously used to evaluate different environmental contaminants. We found that Pb (10μM or higher) was able to inhibit proliferation of exponentially growing cells after 24-h treatment, which was evaluated by the MTT assay and cell counting in Neubauer chamber, but cell survival was not affected according to the trypan blue exclusion assay. On the other hand, Pb was able to increase lipid peroxidation and the number of micronucleated cells, which are indicative of oxidative stress and genotoxic damage respectively. We also found that removal of Pb after 24-h treatment allowed cells to recover proliferation. Our results indicate that Pb was able to induce oxidative stress and genotoxicity in this cell line under standardized conditions, which supports the involvement of Pb in similar effects observed in human exposed to this heavy metal. In addition, Pb inhibits proliferation of exponentially growing fibroblasts but cells resume proliferation after removal of this metal, which suggests that it is important to move away Pb-exposed individuals from the source of contamination.

Yeast Propagation Control: Low Frequency Electrochemical Impedance Spectroscopy as an Alternative for Cell Counting Chambers in Brewery Applications

Electrochemical impedance spectroscopy is a powerful tool in life science for cell and pathogen detection, as well as for cell counting. The measurement principles and techniques using impedance spectroscopy are highly diverse. Differences can be found in used frequency range (β or α regime), analyzed quantities, like charge transfer resistance, dielectric permittivity of double layer capacitance and in off- or online usage. In recent contributions, applications of low-frequency impedance spectroscopy in the α regime were tested for determination of cell counts and metabolic burden in Escherichia coli and Saccharomyces cerevisiae. The established easy to use methods showed reasonable potential in the lab scale, especially for S. cerevisiae. However, until now, measurements for cell counts in food science are generally based on Thoma cell counting chambers. These microscopic cell counting methods decelerate an easy and quick prediction of yeast viability, as they are labor intensive and result in a time delayed response signal. In this contribution we tested our developed method using low frequency impedance spectroscopy locally at an industrial brewery propagation site and compared results to classic cell counting procedures.

Effect of refrigeration at -1°C on spermatozoa quality of domestic cats

ABSTRACT: The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P<0.022) compared to refrigeration at -1°C, using Friedman test. To quantify the decrease in sperm quality, parameter reductions were calculated among time points (F-24h/F-48h/24h-48h). In EPD samples, a greater reduction in sperm quality was detected after 24 hours of refrigeration at 4°C, both in motility and sperm kinetics and in the movement and velocity indices, compared to refrigeration at -1°C. Based on the results, it can be concluded that cooling of feline spermatozoa at -1°C for up to 48 hours was efficient in maintaining spermatic quality collected by EEJ and EPD, and it could be an alternative to spermatozoa cryopreservation in domestic felines.

Achados hematológicos em Iguana iguana (Reptilia, Squamata, Iguanidae) com hemoparasitose em Santarém, Pará, Brasil

A biologia dos répteis pode influenciar diretamente os valores hematológicos, pois alguns parâmetros podem variar de acordo com o sexo, sazonalidade, temperatura, dieta e estado reprodutivo. Na região oeste do estado do Pará, não há informação sobre a presença de hemoparasitas em Iguana iguana e suas possíveis alterações hematológicas. Devido a essa necessidade, objetivou-se identificar a presença de hemoparasitos e as alterações hematológicas provocadas por esses em I. iguana no município de Santarém/PA, Brasil. Foram utilizados 28 espécimes, 13 machos e 15 fêmeas, da cidade de Santarém/PA, Brasil. A pesquisa de hemoparasitas foi realizada em distensões sanguíneas coradas com corante hematológico e analisadas ao microscópio de luz em aumento de 1.000x. Os valores hematológicos foram obtidos por contagem em câmara de Neubauer utilizando o reagente de azul de toluidina 0,01%. Foi observada a infecção por hemoparasitas em 18 animais, 10 machos e oito fêmeas. Não houve diferenças estatísticas significantes entre os sexos e nos valores de leucócitos, trombócitos e hemácias. Existe infecção por hemoparasitas em I. iguana na região. A relação entre este hemoparasita e hospedeiro aparenta ter pouca patogenicidade em provocar alterações importantes nos valores de leucócitos, hemácias e trombócitos.

Anticancer properties of the fatty acid synthase inhibitor TVB-3166 on oral squamous cell carcinoma cell lines

ObjectiveFatty acid synthase (FASN) is overexpressed in several human cancers, including oral squamous cell carcinoma (OSCC). TVB-3166 is a recently described FASN inhibitor with antitumor effects and potential clinical relevance. The objective of this study was to evaluate the effects of TVB-3166 on OSCC cell lines. Materials and methodsThe OSCC cell line SCC-9 modified to express ZsGreen (ZsG) (SCC-9 ZsG) and its in vivo selected metastatic derivative LN-1A were used to evaluate anticancer properties of TVB-3166. Cell viability was determined using MTT assays and proliferation determined by cell counting in a Neubauer chamber. Cell death and cell cycle progression were analyzed by Annexin V-PE/7-ADD-PerCP labeling and PI staining, respectively. Cell migration was assayed by scratch assays and cell adhesion using myogel. Production of FASN, p-AKT, CPT1-α, and epithelial-mesenchymal transition (EMT) markers were examined by Western blotting. ResultsTVB-3166 significantly reduced cell viability and proliferation, promoted cell cycle arrest and apoptosis, and increased adhesion to myogel in both OSCC cell lines. Finally, the drug reduced SCC-9 ZsG migration. ConclusionOur results demonstrated that TVB-3166 has anticancer effects on both SCC-9 ZsG and its metastatic version LN-1A, which are worthy of investigation in preclinical models for OSCC.

A new intraocular lens with artificial iris for treating a case of iris extrusion secondary to traumatic opening of a radial keratotomy

Traumatic globe rupture can lead to aniridia with subsequent glare, light sensitivity and psychological discomfort. The authors report the results of a sulcus implantation of a new intraocular lens with artificial iris (Reper) in an eye affected by traumatic opening of a radial keratotomy incision with total iris extrusion and subluxated cataract. A case of 56-year-old female affected by aniridia and subluxated cataract in left eye secondary to a traumatic opening of one of 12 radial keratotomy underwent a Reper implantation with three-point transcleral fixation. The following parameters were collected at 1, 3 and 6 postoperative months: best-corrected visual acuity (BCVA), intraocular pressure (IOP), central corneal thickness (CCT), simulated keratometry (Sim K), endothelial cell count (ECC) and anterior chamber depth (ACD). Pupil spread function study was performed to evaluate the high-order aberrations and the quality of vision. BCVA improved from light perception to 20/20 (Snellen fraction) at 6 postoperative months. No IOP postoperative peaks were detected. CCT was 520 μm at 6 postoperative months. Corneal topography showed regularization with symmetrical flattening of the central part and a residual peripheral curvature. No significant reduction of ECC was detected. ACD was stable, superior to 3 mm at each time point of follow-up. Pupil spread function study highlighted that the quality of vision was the same in both eyes at the end of follow-up. Reper is a promising functional and cosmetic solution for the treatment of aphakia and aniridia.

Chemical Characterization and Enzymatic Control of Stickies in Kraft Paper Production.

Paper recycling has increased in recent years. A principal consequence of this process is the problem of addressing some polymeric components known as stickies. A deep characterization of stickies sampled over one year in a recycled paper industry in México was performed. Based on their chemical structure, an enzymatic assay was performed using lipases. Compounds found in stickies by Fourier-transform infrared spectrometry were poly (butyl-acrylate), dioctyl phthalate, poly (vinyl-acetate), and poly (vinyl-acrylate). Pulp with 4% (w/w) consistency and pH = 6.2 was sampled directly from the mill once macrostickies were removed. Stickies were quantified by counting the tacky macrostructures in the liquid fraction of the pulp using a Neubauer chamber before the paper was made, and they were analyzed with rhodamine dye and a UV lamp. Of the two commercial enzymes evaluated, the best treatment condition used Lipase 30 G (Specialty Enzymes & Biotechnologies Co®, Chino, CA, USA) at a concentration of 0.44 g/L, which decreased 35.59% of stickies. SebOil DG (Specialty Enzymes & Biotechnologies®) showed a stickies reduction of 21.5% when used at a concentration of 0.33 g/L. Stickies in kraft paper processes were actively controlled by the action of lipases, and future research should focus on how this enzyme recognizes its substrate and should apply synthetic biology to improve lipase specificity.

Eucalyptol reduces airway hyperresponsiveness in rats following cigarette smoke-exposed

BackgroundCigarette smoke is the major cause of airway inflammatory disease, including airway hyperresponsiveness. Eucalyptol (EUC), also named 1.8-cineole, is a monoterpenoid found in essential oil of medicinal plants, showing several biological effects. Hypothesis/purposeBased in the eucalyptol protective activity in respiratory diseases as asthma, our hypothesis is that eucalyptol is able to reduce the airway hyperresponsiveness and the respiratory mechanic parameters in rats exposed to cigarette smoke. Study designWistar rats were divided into control and cigarettes smoke (CS) groups. CS group was daily subjected to cigarette smoke and treated by inhalation for 15 min/day with EUC (1 mg/mL) or vehicle during 30 days. After treatment, bronchoalveolar lavage (BAL) was collected to analyze the inflammatory profile, and tracheal rings were isolated for evaluation of the airway smooth muscle hyperresponsiveness. Lung function was analyzed in vivo. MethodsThe inflammatory profile was evaluated by optical microscopy performing total (Neubauer chamber) and differential leukocyte count (smear slides stained in H&E). The hyperresponsiveness was evaluated in tracheal rings contracted with potassium chloride (KCl) carbamoylcholine (CCh), or Barium chloride (BaCl2) in presence or absence of nifedipine. The lung function (Newtonian resistance-RN) was evaluated by bronco stimulation with methacholine (MCh). ResultsBAL from CS group increased the influx of leukocyte, mainly neutrophils and macrophages compared to control group. EUC reduced by 71% this influx. The tracheal contractions induced by KCl, CCh or BaCl2 were reduced by EUC in 59%, 42% and 26%, respectively. The last one was not different of nifedipine activity. Newtonian resistance (RN) was also reduced in 37% by EUC compared to CS group. ConclusionEUC reduces the hyperresponsiveness and the airway inflammatory profile, recovering the lung function.

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.

Microfabrication of low-cost customisable counting chambers for standardised estimation of sperm concentration.

Evaluation of sperm concentration is essential for research and procedures involving AI, cryopreservation and sperm quality assessment. Microfabrication technologies have shown tremendous potential for rapid prototyping and fabrication of devices to assist reproduction and fertility research, but such utility has not yet been made available for most reproduction laboratories. The aim of this study was to evaluate the feasibility of using microfabrication techniques to produce counting chambers for estimation of sperm concentration. Zebrafish (Danio rerio) spermatozoa were used as a model for evaluation of functionality of the chambers. These microfabricated enumeration grid chambers (MEGC) were composed of a polydimethylsiloxane (PDMS) coverslip with grid patterns (100 μm×100 μm) and a PDMS base platform to create a known volume with a 10-μm height to restrict the cells to a single layer. The results of cell counts estimated by two of three prototype MEGC devices tested were not significantly different from the control device, a commercially available Makler chamber. The material cost for a MEGC was less than US$0.10 compared with product costs of approximately US$100 for a standard haemocytometer and US$700 for a Makler counting chamber. This study demonstrates the feasibility of microfabrication in creating low-cost counting chambers to enhance standardisation and strengthen interdisciplinary collaborations.

Protective role of neutrophils in periodontal inflammation.

The quantification of neutrophils in blood is done to primarily screen the patients with acute infections. On the other hand, the neutrophils have also shown the antimicrobial activity by allowing the host cells to perform the primary function of preventing the wounds from being infected. The aim of this study was to quantify the blood neutrophil count in patients with chronic gingivitis and chronic periodontitis and to compare with the healthy controls in order to assess the protective role of neutrophils in periodontal inflammation. A total of 30 subjects in the age group of 30-55 years were selected. Based on the clinical examination, the subjects were divided into three groups Group I (healthy), Group II (chronic gingivitis) and Group III (chronic periodontitis). The clinical parameters such as modified gingival index (MGI) and clinical attachment level (CAL) were recorded. Two millilitres of venous blood was collected from each participant in all the groups and the differential neutrophil counts were obtained using the Neubauer's chamber under a light microscope. One way ANOVA test was used to calculate the mean and standard deviation for MGI, CAL and blood neutrophil count among various groups. The mean neutrophil counts in the various study groups were 63.300, 64.100 and 60.800 in Group I, Group II and Group III, respectively. Though there was a decrease in the mean neutrophil count from Group II to Group III, there was no statistically significant difference between the groups. In our study, there was no significant increase in neutrophil count in chronic periodontitis as compared to healthy controls, thereby explaining the protective role of neutrophils in periodontal inflammation.

Seasonal changes in testicular ultrasonogram pixel-intensity and their association with semen characteristics in rams

Objective: To establish reference values for pixel intensity of testicular ultrasonogram of rams in breeding and non-breeding seasons, and to investigate if the seasonal changes in testicular echogenicity and heterogeneity are associated with semen characteristics. Methods: Five Awassi rams aged 3-5 years old and weighing 45-60 kg were subjected to ultrasonographic scanning of the testes twice monthly for one year (from January 2018 to December 2018), together with semen collection and evaluation of ejaculate volume, sperm motility, sperm concentration, sperm morphology and viability. The ejaculate volume was determined in a graduated collection tube (scale of 0.1 mL). Mass and individual sperm motility was expressed in percentage of motile spermatozoa under optical microscope equipped with a warm stage. Sperm concentration was determined by using a Neubauer chamber. Semen smears stained with eosin-nigrosin were used to determine the percentage of live spermatozoa and sperm cell morphology by using a light microscope. Results: The mean testicular pixel intensity was the lowest in winter and increased gradually from breeding season to non- breeding season, reaching its maximum value in summer (P Conclusions: Pixel intensity values of testicular ultrasonogram in rams undergo marked seasonal changes that are associated with fluctuations in photoperiod and ambient temperature. The resulting values of testicular echogenicity (pixel intensity) throughout the year provide useful reference values for predicting the testicular function in Awassi rams.

In vitro EVALUATION OF ATOVAQUONE ON THE REPLICATION OF TOXOPLASMA GONDII TACHYZOITES STRAINS RH AND ME49

Introduction: Since 1990 the standard treatment for toxoplasmosis has remained the same. The therapy is based on a synergistic combination of pyrimethamine and sulfadiazine, drugs that promote synthesis blockade and decrease of tachyzoite folic acid levels. This therapy acts only on the tachyzoite forms of the parasite, not affecting bradyzoites present in tissue cysts that persist in the individual during the chronic phase and not preventing possible disease reagudization even after treatment. Given the high seroprevalence of toxoplasmosis in the general population and the serious complications that this infection may bring to the patient, a safe and effective treatment against all morphological forms is necessary. The repositioning of drugs main objective is to use substances already marketed to treat other diseases. Thus, Atovaquone, which is an antimalarial of the naphthoquinone group recently accepted by the FDA and isn’t yet part of the routine treatment protocols employed for toxoplasmosis, should be studied, as recent studies show its activity against tachyzoites and also against bradyzoites, representing a huge advantage over the drugs currently used. It also does not interfere with folic acid metabolism, proving to be a promising drug in the treatment of pregnant women. Objective: The present study aims to evaluate the effects of atovaquone on T. gondii strains RH and ME49 (type I and type II, respectively) in vitro. Methodology: In quintuplicate, murine RAW 264.7 macrophages were used in six-well culture plates with 3mL of supplemented RPMI medium, where 200.000 cells were seeded and incubated at 37°C, 5% de CO2 for 24 hours. After this period, the culture was infected with 1×106 tachyzoites/well of each strain analyzed and simultaneously treated with 100 nM Atovaquone. The parasites were analyzed by optical microscopy and quantified in a Neubauer chamber at 24h, 48h, 72h and 7 days. Results: For the RH strain, the control group showed steady growth, starting at 60.000 forms/mL within 24 hours, reaching 2.550.000/mL within 7 days. When treated with Atovaquone, within 24 hours the number detected was 30.000/mL, with decline in growth resulting in only 15.000/mL observed after the first week of cultivation. Similar results were observed with the Me49 strain, which at the beginning of the experiment had 150.000/mL tachyzoites in the untreated group and 90.000/mL in the treated group and, at the end of 7 days, 1.950.000/mL tachyzoites in the untreated group and 30.000/mL in the treated group. Conclusion: Atovaquone has shown great activity in controlling Toxoplasma gondii parasite replication in vitro. Financial Support: FAPEG (Fundacao de Amparo a Pesquisa do Estado de Goias)

Validation of a new method for estimating low platelet counts: G&S method

ABSTRACT Introduction: Automated hematology analyzers are able to produce low platelet counts with great precision and accuracy. However, these analyzers may produce erroneous counts due to the presence of interferences. Therefore, additional methodologies are required to confirm platelet counts, such as blood smear and a direct count, using the Neubauer chamber. Objective: To validate the reliability estimate produced by the G&S method. Material and method: One hundred and fifty platelet counts were analyzed in the hematology and emergency laboratories of the Hospital Nacional Docente Madre-Nino San Bartolome, in Lima, Peru, by four methodologies: one optical platelet count (PLTO), one indirect blood smear count (Indirect), and two direct counts (Direct and G&S). Descriptive and inferential statistics were used to compare the groups. Results: A significant difference was observed in the distribution of the four methodologies and, after applying the post-hoc analysis, a similarity between the PLTO and G&S methods was found. Also, the Direct, G&S and Indirect methods showed a positive correlation with the PLTO method. The Bland-Altman test demonstrated that the G&S method presented a high agreement with the PLTO method. Conclusion: The G&S method is inexpensive, easy to perform, and has demonstrated statistical reliability concerning the automated methodology, and is useful for confirming low platelet counts after a suspected error by the automated equipment or when this device is not available for use.