Background: The purpose of tissue processing is to strengthen the tissue and place it in a suitable environment for slicing without causing damage. In routine histopathology, decalcification of calcified tissues is a principal step before tissue processing. Thus, the aim of this study is to evaluate the effect of several decalcifying agents on morphological and antigenic preservation in canine mandibular tissue. Methods: Four different decalcifying solutions, including 5% nitric acid (NA), 10% and 20% formic acid (FA), and 10% ethylene diamine tetraacetic acid (EDTA), were employed to remove mineral salts from specimens harvested from mandibular osseous tissues of four healthy Mongrel dogs (32 samples). The solutions were compared regarding the decalcification time, ease of tissue slicing, morphological organization after Alizarin red S staining, and osteocalcin antigenic preservation. Results: Considering the short decalcification time and ease of sectioning, 5% NA) yielded better results than 10% and 20% FA and 10% EDTA (pH 7.4). In terms of the preservation of morphology and antigenicity of the tissue samples, 10% EDTA was found to be the most optimal solution, followed by 10% and 20% FA and 5% NA. Conclusion: Our findings support EDTA as a highly preferred choice for the decalcification of canine osseous tissue when aiming for immunohistochemistry, despite its time-consuming nature. However, for general histological staining procedures, 20% FA and 5% NA are preferred.

Major biological processes rely on the spatial organization of cells in complex, highly orchestrated three-dimensional (3D) tissues. Until the recent decade, most of information on spatial neural representation primarily came from microscopic imaging of “ 2D” (5-50 μ m) tissue using traditional immunohistochemical techniques. However, serially sectioned and imaged tissue sections for tissue visualization can lead to unique non-linear deformations, which dramatically hinders scientists’ insight into the structural organization of intact organs. An emerging technique known as CLARITY renders large-scale biological tissues transparent for 3D phenotype mapping and thereby, greatly facilitates structure-function relationships analyses. Since then, numerous modifications and improvements have been reported to push the boundaries of knowledge on tissue clearing techniques in research on assembled biological systems. This review aims to outline our current knowledge on next-generation protocols of fast free-of-acrylamide clearing tissue (FACT) and passive CLARITY (PACT). The most important question is what method we should select for tissue clearing, FACT or PACT. This review also highlights how FACT differs from PACT on spanning multiple dimensions of the workflow. We systematically compared a number of factors including hydrogel formation, clearing solution, and clearing temperatures between free-acrylamide and acrylamide-based passive sodium dodecyl sulfate (SDS) tissue clearing and discussed negative effects of polyacrylamide on clearing, staining, and imaging in detail. Such information may help to gain a perspective for interrogating neural circuits spatial interactions between molecules and cells and provide guidance for developing novel tissue clearing strategies to probe deeply into intact organ.

Introduction: The current study aimed to identify and characterize primordial germ cells in the blood, genital ridge, and primary gonad in turkey. Besides, we described the histological characteristics of ovaries in the turkey embryo. Methods: The embryos from stages 14 to 31 per Hamburger and Hamilton, were studied by the means of blood smear and serial sections from the whole-mount embryo. The primordial germ cells were identified by histochemical and immunostaining techniques. Results: The present research results suggested that these cells could be identified by their remarkably large size, large nuclei, and granules in the cytoplasm. At stages 20-21 of Hamburger and Hamilton, the alkaline phosphatase reaction was negative in these cells. Furthermore, the primordial germ cells could not be labeled by stage-specific antigen-1 antibody in the primary gonad. We observed that these cells provided negative or poorly positive staining with Alcian blue solution in the migration phase. The presence of glycogen in the cytoplasm of the primordial germ cells was verified using periodic acid-Schiff and Best’, s Carmine methods. These cells were Best’, s Carmine positive,however, they contained reduced amounts of glycogen in the primary gonad. Conclusion: The study findings demonstrated that the periodic acid-Schiff is the best method for identifying the turkey primordial germ cells in the blood and migration phase. Moreover, we reported the existence of histological differences between the right and left ovaries in the turkey embryo.

Background and Aim: Diagnosis of dental caries with Diagnodent device has been found to be comparable to visual and radiographic diagnostic methods. The aim of this study was to determine the effect of Aging, Staining, and Bleaching on Diagnodent readings in composite resin restorations.Materials and Methods: This was an experimental in vitro study in which 108 disk-shaped specimens were fabricated from 6 different composite resins including: 3 M p60, Kerr point 4, Vivadent Heliomolar, Ultradent flow, Tetric flow and Kerr flow. The samples were divided into 3 groups. Group 1was aged in water, group 2 was stained with tea and group 3 was bleached with 16% Carbamide peroxide. Specimen Fluorescence was measured with Diagnodent (Kavo- U.S) before and after these treatments. Data were analyzed using mixed repeated Measure Model ANOVA.Results: This study was performed on 108 composite resin specimens. All samples aged in water showed higher reading values; only Tetric flow exhibited a lower mean value (P<0.001). All samples stained with tea showed higher readings(P<0.001) and all materials bleached with 16% Carbamide peroxide were reported to have lower values; only Kerr flow exhibited a higher mean value than basic situation.Conclusion: The results indicate that aging and staining can result in an increase in the Diagnodent readings of the composite resin restorations while bleaching lead to a decreased the readings. However, the reported differences in Diagnodent readings were not significant enough to result in a false positive diagnosis of caries.

Introduction: Semen analysis including sperm count, progressive and non-progressive motility, and normal morphology of spermatozoa of ejaculates from infertile men are important. In general, assisted reproductive techniques (ART) are chosen according to the semen parameters. Therefore, it is crucial to evaluate semen parameters with application of one quick, standard, and inexpensive methodology. In some ART laboratories in Iran, the aforementioned analysis is done using traditional methodology such as wet preparation (microscopic slide technique) which can mislead the infertile couple. As a result, the infertile couple may not fully benefit from the treatment outcome. The objective of this prospective study was to compare the two techniques of Wet preparation and Makler - staining for analysis of semen parameters. Materials & Methods: The three parameters of count, morphology and motility of spermatozoa along with round cells of ejaculates from 50 men attending the infertility treatment program were evaluated. Following macroscopic evaluation, each seminal sample was evaluated with two different techniques of Wet and Makler - staining in a blind setting. For wet slide, semen parameters as well as round cells were studied. However, for Makler technique, Makler chamber was used for evaluation of percentage of motility and sperm count. In addition, Geimsa (WHO) staining was applied for evaluation of sperm morphology and round cells. The results showed that the volume, PH and viscosity of semen were within normal range of WHO. Results: The difference in the mean sperm count & percentage of motility was statistically insignificant. However, the sperm morpology was significantly different in the aformentioned groups (P=0.001). Also, the mean number of round cells in both groups was different (P=0.002). A total of one and nine seminal specimens were clear from round cells in Wet and Makler groups, respectively. The results showed that most variation was related to sperm morphology. For example, 5 seminal samples were abnormal in the wet method, while 32 ejaculates were abnormal in the Makler group. Conclusion: Sperm morphology' and round cells should be evaluated with more attention. Sperm morphology plays an important role in ART methodology and therefore should be counted and detected in full. It should be noted that Wet preparation technique may be used for evaluation of concentration and motility if Makler chamber is unavailable at ART laboratories.    

A difference labeling of a graph G is an injective function f: V (G) → N ∪ {0} together with the weight function f∗ on E(G) given by f∗(uv) = |f(u)-f(v)| for every edge uv in G. The collection of subgraphs induced by the edges of the same weight is a decomposition of G and is called the common weight decomposition of G induced by f. Let ϒf denote the collection of all the paths taken from each member of the common weight decomposition induced by f. A difference labeling f of G is said to be a graphoidal difference labeling if ϒf is an acyclic graphoidal decomposition of G. This paper initiates a study on this concepts.

PCR involves a repeating cycle of replication to amplify small segments of deoxyribonucleic acid (DNA). A novel application of this technique is microbial identification in infectious keratitis, one of the leading causes of blindness in the world. PCR is more sensitive than biological stains and culture, which are considered the current gold standards for diagnosing infectious keratitis. The diagnosis and treatment of infectious keratitis cost the United States millions of dollars in health expenditure. PCR may help offset that cost by allowing for individualized disease management and screening for multiple antibiotic-resistant genes. While beneficial, PCR demonstrates lower specificity rates compared to culture and stain, indicating its shortcomings; this can be overcome by performing PCR after narrowing the pool of potential microorganisms. This article examines the clinical utility of PCR in cases of infectious keratitis by evaluating its reliability, validity, associated costs, and indications. . .

Background: Reticulocytes are immature red blood cells with RNA, spending the final stages of their maturation in the peripheral blood. The number of reticulocytes in the peripheral blood is the salient evidence of the effectiveness of bone marrow to produce red blood cells. Currently, reticulocyte count is done manually or automatically in clinical laboratories. Difficulties and limitations of these approaches, including imprecision, poor reproducibility and laboriousness are clear. This study used modification in manually staining method to achieve better counting. Materials and Methods: The reticulocyte counts of 30 samples were obtained from patients referred to Afzalipour hospital in Kerman, Iran. The patients aged between 0-3 years old (median age= 1. 4 years). Samples were investigated for reticulocyte count by three different methods, including conventional, modified, and flow cytometry methods. Modified staining method is used in the optimized method which helps operator to countg reticulocyte better than conventional method. Results: Analysis of the results showed a reasonable agreement. Sphericity assumption was checked by Kolmogorov-Smirnov and Mouchley analyses. Repeated measurement analysis was performed using Greenhouse-Geisser correction (F=16. 6, df=1. 105) according to pairwise comparisons with Bonferroni adjustment. Mean differences for flow cytometry, optimized and common methods were 3. 88, 3. 59, and 2. 46 were significant at the 0. 05 level, respectively. Conclusion: The proposed optimized method was simple method, highly reliable, and comparable to flow cytometry. Due to the ease of implementation, this method did not need specialized training or the use of complex devices so it is affordable.