Staining Characteristics Research Articles

Effects of Erchen Decoction on Oxidative Stress-Related Cytochrome P450 Metabolites of Arachidonic Acid in Dyslipidemic Mice with Phlegm-Dampness Retention Syndrome: A Randomized, Controlled Trial on the Correspondence between Prescription and Syndrome.

Phlegm-dampness retention (PDR) syndrome is one of the main syndromes of dyslipidemia. This study investigated the effects of Erchen decoction (ECD) on concentrations of two oxidative stress-related cytochrome P450 (CYP450) metabolites of arachidonic acid—14,15-dihydroxyeicosatrienoic acid (14,15-DHET) and 20-hydroxyeicosatetraenoic acid (20-HETE)—in mice with dyslipidemia and phlegm-dampness retention (PDR) syndrome (n = 5 C57BL/6J mice and n = 30 apolipoprotein E knockout mice). Murine models of the disease and syndrome were established using multifactor stimulation. Then, all mice were assigned to normal, model, low- (L-), medium- (M-), or high- (H-) dose ECD groups or to a control or an unmatched prescription-syndrome (unmatched P-S) group; five mice were included in each group. Dose formulations were administered by oral gavage for 30 days to animals in the corresponding groups. We detected and analyzed hematoxylin and eosin (HE) staining characteristics of the mouse aorta and serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), peroxynitrite (ONOO−), 14,15-DHET, and 20-HETE concentrations in each group. TC and LDL-C concentrations significantly decreased in the M-ECD versus control group (P < 0.05); however, the TC and LDL-C concentrations were not significantly different in the unmatched P-S versus model group (P > 0.05). After treatment in the P-S correspondence groups (L-ECD, M-ECD, and H-ECD groups), the concentration of ONOO− decreased to different degrees in each group. Among these groups, the concentration of ONOO− significantly decreased in the L-ECD, M-ECD, and H-ECD groups versus the model group (P < 0.05). However, the concentration of ONOO− was not significantly different in the unmatched P-S versus the model group (P > 0.05). From the perspective of aortic HE staining, the P-S group experienced an improved endothelium structure after treatment. 14,15-DHET concentrations significantly increased in the normal, M-ECD, and H-ECD groups versus the model group; in the H-ECD versus the L-ECD group; and in the H-ECD versus the control group (all P < 0.05) to various extents after different doses of the prescription. 20-HETE concentrations pronouncedly decreased in the M-ECD versus normal group; in the M- and H-ECD groups versus the model group; in the M-ECD versus the L-ECD group; in the M-ECD versus the control group; and in the M-ECD versus the unmatched P-S (P < 0.05). However, the concentrations of 14,15-DHET and 20-HETE in the model group were not significantly different from those of the unmatched P-S (P > 0.05). In this study, ECD reversed blood lipid indexes and ameliorated oxidative stress-related metabolites, elevating serum 14,15-DHET and lowering serum 20-HETE in mice with dyslipidemia and PDR syndrome via CYP450 pathways of arachidonic acid metabolism. The efficacy of ECD relies on correspondence between the prescription and the syndrome. These findings scientifically validate the treatment according to traditional Chinese medicine syndrome differentiation. ECD can strengthen the protective effect on the vascular endothelium by driving out pathogenic factors and strengthening healthy resistance. Its efficacy may be related to the adjustment of the polarization state of macrophages.

Staining susceptibility of dental composite resins with various nano-filler technologies

Background: Discoloration of dental composite restoration is a common problem that usually results in refurbishing or replacement or even removal of the restoration. Objective: The purpose of this study was to evaluate the color stability of polished composite resin material with different filler technologies. Methodology: Three composites were studied; Filtek™ Z350 XT (3M, ESPE, St. Paul, USA) [Group-I] that has nano-cluster filler particles, Brilliant EverGlow™ (Coltene/Whaledent® AG, Altstatten, Switzerland) [Group-II] which consists of submicron barium glass fillers and pre-polymerized fillers and Ceram.X® Sphere TEC™ (Dentsply, Konstanz, Germany) [Group-III] with advanced granulated filler technology composite. Twenty standardized composite discs were prepared with each composite resin, and it was polished with Sof-Lex disks (3M, ESPE, St. Paul, USA). The baseline color was then recorded using a spectrophotometer (X-Rite PANTONE® iPro-2, Michigan). Eight samples from each group were then subdivided and immersed into freshly prepared solutions of turmeric, coffee, and four samples into distilled water for a total period of 3 hours/day for 30 days. Following this, the color was again recorded. Statistical analysis: The change in color (ΔE) was calculated, and the data obtained were subjected to Kruskal Wallis and Mann Whitney test. Results: A significant difference was seen between the staining characteristics of Group-III and Group-I (p&amp;lt;0.05). The mean color change in the values was highest in Ceram.X that was clinically unacceptable (ΔE&amp;gt;3.3). Conclusion: Though all composites revealed color changes after their immersion into the staining solutions, the amount of stain varied based on their constituents and filler characteristics.

Clinical, Morphological and Histopathological Staining Characteristics of Mucormycosis in COVID 19 Patients

COVID 19, caused by severe acute respiratory syndrome – corona virus -2 (SARS – CoV 2), is a pandemic that affected millions of people worldwide. During the second wave of COVID -19 pandemic in India, an unprecedented surge in cases of mucormycosis was observed. This study was carried out to describe the clinical, morphological and histopathological staining characterstics of mucor mycosis in 16 covid patients in a tertiary care setting. Older patients with comorbidities like diabetes mellitus are commonly affected. The most common site of involvement was sinonasal tract (68.75%) followed by sinonasal tract and orbit (37.5%). Out of the 16 patients, 2 were partially vaccinated whereas others were not vaccinated. Histopathological examination of specimens revealed broad aseptate fungi in all cases. Tissue necrosis was present in all cases. Suppuration being the most common tissue reaction. 43.75% showed angioinvasion and 37.5% had bone invasion. All the cases were GMS positive with variable intestity. Treatment given in all cases were liposomal amphotericin B injection and surgical wound debridement. On follow up, 40 % of the patients died inspite of aggressive clinical management. Hence this fatal fungal infection should be kept in mind in covid 19 recovered patients especially those who are immunocompromised.

Polymicrobial oral conventionalization model in mice.

The aim of this study is to describe a new polymicrobial oral conventionalization protocol in mice. Oral biofilm samples were collected from wild C57 BL/6 mice (WG), which had not been previously submitted to any experimental procedure. The contents of these samples were used for inoculation in the oral cavity of specific pathogen free (SPF) animals. This inoculation was repeated 3 times. Qualitative cytological analyses were performed in the days 0, 16 and 80 of the experimental protocol, to check the presence or absence of microorganisms, their morphology and staining characteristics on the oral cavity of the animals. At the end of this study, was observed a combination of oral bacterial microbiota of SPF animals and wild animals in the conventionalized group (CONV). Samples collected from CONV mice on day 16, a period in which these animals had been previously inoculated 3 times with wild mouse microbiota, showed a greater amount of Gram-positive cocci, as seen in SPF animals. In addition, Gram-negative cocci were present, although in a much smaller proportion than previously seen in wild mice. On the 80th experimental day, CONV animals showed a predominance of Gram-positive cocci and bacilli. Filamentous bacteria were also seen in this group. The conventionalization of SPF animals using the technique with inoculum from the resident microbiota of wild mice proved to be an effective, low-cost and easily reproducible technique. The conventionalized animals showed the colonization of a microbiota similar to wild animals up to 80days of experiment.

Therapeutic efficacy of adipose-derived mesenchymal stem cells after chronic fluoxetine treatment on pars distalis in adult male albino rats

BackgroundFluoxetine hydrochloride is one of the most commonly used antidepressants in the selective serotonin reuptake inhibitor class. The aim of work: The study was conducted to detect the effect of chronic fluoxetine treatment on pars distalis and the possible therapeutic effect of adipose-derived mesenchymal stem cells (ADSCs). Material and methodThirty healthy male adult albino rats were classified into four groups. Control group (Group I) included fifteen rats. Fluoxetine treated (Group II) included five rats that received 24 mg/kg/day of fluoxetine dissolved in 1.0 ml of tap water once a day for 30 days. Fluoxetine group treated with ADSCs (Group III) included five rats that received fluoxetine as group (II) for 30 days, then supplied once by ADSCs at a dose of 1 × 106 cells/rat in the tail vein suspended in 0.5 ml of phosphate-buffered saline (PBS). Recovery group (Group IV) included five rats that received fluoxetine as the group (II) then received no treatment till the end of the experiment. Samples from pars distalis were processed for light, electron microscopic examination, morphometrical and statistical analyses. ResultsIn the fluoxetine-treated group, there was a disruption in cellular architecture, size, shape, and staining characteristics of pars distalis cells. The administration of ADSCs significantly improved the microscopic appearance of cells, while the recovery group showed some histological changes similar to the fluoxetine group. ConclusionFluoxetine induced various deleterious changes in the pars distalis of albino rats. These changes were almost corrected by the ADSCs treatment.

Assessing malaria staining practices with Giemsa for malaria microscopy; a case of selected laboratory facilities in the Ashanti region of Ghana

BackgroundMicroscopy remains the gold standard for malaria diagnosis globally, especially where quality Giemsa stain is used. However, several Giemsa stains products have flooded the Ghanaian market which may not meet the required standard. In this study we report on the efficacy of Giemsa stains used for malaria detection in selected medical laboratories in Ashanti region. MethodologyA questionnaire was administered to obtain data of staining practices among participants. Thick and thin blood smears with standard parasite density of 300,000 parasites/ul ± 25,000 was given to facilities for staining. The stained slides were examined for their staining characteristics of the white blood cells, malaria parasites and parasite density by certified malaria microscopists. ResultsLess than 40% of the participant had parasite density within the expected range (225,000-375,000/ul). Forty percent (40%) also scored less than 7 out of 10 on the staining characteristics of the white blood cells. Additionally, 73.3% of participating laboratories used tap water for preparation of Giemsa working solution which affected parasite negatively. A statistically significant association was observed between immediate preparations of working solution and the morphological changes of neutrophils (p<0.0001). ConclusionThe study found that a less than half (40%) of Giemsa products used in Ghanaian medical laboratory facilities were adequately effective in detecting in malaria parasites optimally. This poses a great threat to malaria diagnosis in Ghana.

Histopathological Analysis of Central Nervous System Metastases: Six Years of Data From a Tertiary Center.

Introduction: The most common cause of neurological symptoms in patients with systemic malignant tumors is central nervous system (CNS) metastases, and CNS metastases are one of the important causes of morbidity and mortality in these patients. The most common metastatic tumors to the CNS are lung, breast, malignant melanoma, genitourinary, and gastrointestinal tumors. We aimed to analyze our data on patients with CNS metastases in our department, which belongs to a large archive in the field of neuropathology.Methods: The data of patients who had CNS metastases between January 2015 and August 2021 in our department were reviewed retrospectively. The patients were grouped in terms of demographic data, location, histopathological diagnosis, and primary origin characteristics, and their frequency and immunohistochemical staining characteristics were investigated.Results: There were 256 patients with CNS metastases in our study. The mean age was found to be 56.12. Of the patients, 30.5% were female and 69.5% were male. Astrocytic and oligodendral tumors (25.3%), followed by meningiomas (24.1%), and then CNS metastases (21.3%) were the most common CNS tumors. Among the CNS metastases, the most common primary sites were the lung (58%), breast (16%), tumors of unknown primary origin (TUP) (5%), colon (4%), and gynecologic tract (3.1%). Localization was found as cerebral (69.5%), cerebellar (28.1%), and spinal (2.3%).Conclusion: In CNS system metastases, an accurate histological diagnosis should be made by histomorphological evaluation supported by compatible immunohistochemical results in the presence of clinical history and radiological findings. Despite performing a larger immunohistochemical panel, it should be kept in mind that a primary site of origin cannot be found in a significant number of cases.

Immune tumoral microenvironment in gliomas: focus on CD3+ T cells, Vδ1+ T cells, and microglia/macrophages.

Gliomas are histologically defined as low-grade gliomas (LGG) and high-grade gliomas (HGG). The most common type of HGG is the glioblastoma (GBM). We aimed to determine the immunological characteristics of CD3 T-cells, Vδ1 T-cells, and microglia/macrophages infiltrating brain gliomas. We collected 24 formalin-fixed paraffin-embedded samples issued from 19 cases of GBM and 5 cases of LGG. An immunohistochemical analysis was performed using anti-CD3, anti-Vδ1, and anti-iba-1 antibodies. Labelling indexes (LI) of CD3 and Vδ1 were evaluated quantitatively, and other CD3, Vδ1, and iba-1 staining characteristics were evaluated qualitatively. The median age of patients was 49years in GBM and 52years in LGG. The sex ratio was 1.4 and GBM predominated in males (p = 0.05). In GBM, the medians of CD3-LI and Vδ1-LI were 30 and 3.5 respectively. CD3-LI inversely correlated with survival in GBM cases (r = - 0.543; p = 0.016). CD3 staining intensity correlated with CD3-LI (p < 0.0001) and with the survival in GBM cases (p = 0.003). Compared to LGG, the CD3-LI, the intensity of intra-tumoral Vδ1 staining, and the amount of iba-1 were higher in GBM (p = 0.042; p = 0.014; and p = 0.001 respectively). The iba-1 organization was more amoeboid in older patients and more branched in younger patients (p = 0.028) and tended to be more amoeboid in cases with high iba-1 amount (p = 0.09). Our results suggest that a high level of CD3-LI and a strong intra-tumoral infiltration of Vδ1 T-cells as well as a high involvement of TAM can be considered potential markers of poor prognosis of GBM. However, this requires further studies on more balanced GBM-LGG sample, including an expanded panel of biomarkers.

Prior Incubation as a Tool to Overcome Understaining of Leishman Stain: A Cross-sectional Study

Introduction: A well stained peripheral smear is an integral part in the diagnosis and management of many clinicohematologic conditions. Leishman stain is recommended universally by the International Committee for Standardisation in Haematology (ICSH). However, a commonly encountered problem in tropical countries, especially during the rainy season, is under staining by Leishman stain as a result of interference by the atmospheric moisture. Hence, incubation of slides and/or buffer prior to standard Leishman staining protocol could bring out best staining qualities. Aim: To assess the effects of prior incubation of slides and/or buffer on standard Leishman staining technique. Materials and Methods: The cross-sectional study was conducted over a period of 8 weeks in the Central Laboratory, Kempegowda Institute of Medical Sciences, Bangalore in the months of October 2020 to November 2020. Blood samples were received in the central laboratory for routine haematological investigations. Total of 100 samples of left over non haemolysed Dipotassium Ethylenediaminetetraacetic Acid (K2-EDTA) anticoagulated blood irrespective of age, gender and cell counts were included in the study. Slides were stained following prior incubation of slides and/or buffer at 37° C and compared with the standard staining technique. The staining characteristics of nucleus, cytoplasm and granules of White Blood Cells (WBCs), platelets and erythrocytes were observed. One-way Analysis of Variance (ANOVA) test and p-value of &lt;0.05 was considered statistically significant and higher f value (&gt;104.35152) rejected the null hypothesis. Results: Prior incubation of slides and/or buffer resulted in better stained smears which were found to be statistically significant (p-value&lt;0.05) as compared to standard Leishman staining technique. Conclusion: Minor modifications when applied to standard Leishman staining technique such as prior incubation of slides and/ or buffer at 37° Centigrade produced best staining characteristics to overcome the effects of humidity which normally interfere with staining of slides.

An expandable informatics framework for enhancing central cancer registries with digital pathology specimens, computational imaging tools, and advanced mining capabilities

BackgroundPopulation-based state cancer registries are an authoritative source for cancer statistics in the United States. They routinely collect a variety of data, including patient demographics, primary tumor site, stage at diagnosis, first course of treatment, and survival, on every cancer case that is reported across all U.S. states and territories. The goal of our project is to enrich NCI’s Surveillance, Epidemiology, and End Results (SEER) registry data with high-quality population-based biospecimen data in the form of digital pathology, machine-learning-based classifications, and quantitative histopathology imaging feature sets (referred to here as Pathomics features). Materials and methodsAs part of the project, the underlying informatics infrastructure was designed, tested, and implemented through close collaboration with several participating SEER registries to ensure consistency with registry processes, computational scalability, and ability to support creation of population cohorts that span multiple sites. Utilizing computational imaging algorithms and methods to both generate indices and search for matches makes it possible to reduce inter- and intra-observer inconsistencies and to improve the objectivity with which large image repositories are interrogated. ResultsOur team has created and continues to expand a well-curated repository of high-quality digitized pathology images corresponding to subjects whose data are routinely collected by the collaborating registries. Our team has systematically deployed and tested key, visual analytic methods to facilitate automated creation of population cohorts for epidemiological studies and tools to support visualization of feature clusters and evaluation of whole-slide images. As part of these efforts, we are developing and optimizing advanced search and matching algorithms to facilitate automated, content-based retrieval of digitized specimens based on their underlying image features and staining characteristics. ConclusionTo meet the challenges of this project, we established the analytic pipelines, methods, and workflows to support the expansion and management of a growing repository of high-quality digitized pathology and information-rich, population cohorts containing objective imaging and clinical attributes to facilitate studies that seek to discriminate among different subtypes of disease, stratify patient populations, and perform comparisons of tumor characteristics within and across patient cohorts. We have also successfully developed a suite of tools based on a deep-learning method to perform quantitative characterizations of tumor regions, assess infiltrating lymphocyte distributions, and generate objective nuclear feature measurements. As part of these efforts, our team has implemented reliable methods that enable investigators to systematically search through large repositories to automatically retrieve digitized pathology specimens and correlated clinical data based on their computational signatures.

Deep learning-based bacterial genus identification.

This study aimed to develop a computerized deep learning (DL) technique to identify bacterial genera more precisely in minimum time than the usual, traditional, and commonly used techniques like cultural, staining, and morphological characteristics. A convolutional neural network as a part of machine learning (ML) for bacterial genera identification methods was developed using python programming language and the Keras API with TensorFlow ML or DL framework to discriminate bacterial genera, e.g., Streptococcus, Staphylococcus, Escherichia, Salmonella, and Corynebacterium. A total of 200 digital microscopic cell images comprising 40 of each of the genera mentioned above were used in this study. The developed technique could identify and distinguish microscopic images of Streptococcus, Staphylococcus, Escherichia, Salmonella, and Corynebacterium with the highest accuracy of 92.20% for Staphylococcus and the lowest of 77.40% for Salmonella. Among the five epochs, the accuracy rate of bacterial genera identification of Staphylococcus was graded 1, and Streptococcus, Escherichia, Corynebacterium, and Salmonella as 2, 3, 4, and 5, respectively. The experimental results suggest using the DL method to predict bacterial genera included in this study. However, further improvement with more bacterial genera, especially of similar morphology, is necessary to make the technique widely used for bacterial genera identification.

Impact of Gamma Irradiation on Physicochemical, Technological, Antioxidant and Microbiology Properties of Whole Sorghum Flours

This study aimed to evaluate the effect of gamma irradiation (GI) on the physicochemical, technological, antioxidant and microbiological characteristics of different whole sorghum flours (WSF), as well as to characterize the profile of chemical constituents by paper spray mass spectrometry (PS-MS). The doses applied interfered in the staining characteristics of the evaluated flours, providing the darkening of the same. For the other physicochemical, technological, and antioxidant parameters, no significant influence of the process was observed. The fingerprint obtained in both ionization modes had not been influenced by irradiation, being flavonoids, phenylpropanoids, amino acids, benzoic acid derivatives, carboxylic acids, and sugars tentatively identified. In microbiological terms, there was a reduction of molds, yeasts, and Bacillus cereus in irradiated WSF compared to control (non-irradiated). Therefore, the application of low doses of gamma irradiation represents an advantageous alternative for the conservation of WSF and maintenance of compounds bioactive identified by the PS-MS technique.

Grape Juice as Alternative Agent for Xylene in Hematoxylin and Eosin Staining Procedure

Abstract: Aim: To compare the grape juice as substitute for xylene agent in hematoxylin and eosin staining procedure. Objective: The objective is to find the use of grape juice as deparaffinizing agent as a substitute to xylene and comparing the staining characteristics in grape juice and Xylene as deparaffinising agent in hematoxylin and eosin stain. Materials and Methods: The study made in 45 paraffin embedded blocks of breast fibroadenoma tissues. From each block, two sections of 4-5 microns thickness were prepared. Two different groups like A and B were seperated. Tissue sections in Group A were stained with H &amp; E method where xylene was used as deparaffinizing agent and Group B were stained with H &amp; E with grape juice, were it used as deparaffinising agent alternative to Xylene. Staining characteristics were compared with xylene and scoring was given. The scoring was given to the tissue depending upon the staining of the cells, 3–5 score was regarded as satisfactory for diagnosis and less than that is insufficient for diagnosis. Results: Impression obtained with staining characteristics in the cell eg : nuclear, cytoplasm, uniformity, clarity of the cell and crispiness of staining for diagnosis was greater with grape juice than xylene. Conclusion: The natural method can be used as deparaffinizing agent such as grape juice an alternative to xylene.

Comparison of Efficacy of the Pap and H &amp; E in Identifying the Various Changes in Liver Samples of High Fat Diet Induced Rat - In vitro Study

Introduction: High fat induced (HFI) liver is associated with a high calorie intake and many other diet-induced complications, such as metabolic syndrome and cardiovascular disease. HFI is the most common chronic liver disease that develops due to high fat intake. A wide range of liver injuries is associated with HFI, ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), advanced fibrosis and cirrhosis. The signs and symptoms of metabolic syndrome may be induced in rats by feeding them a diet rich in carbohydrates and fat. The aim of the study is Comparison of efficacy of the PAP (Papanicolaou) stain and H&amp;E (Hematoxylin and eosin) stain in identifying the various changes in liver samples of high fat diet induced rat.&#x0D; Materials and Methods: Liver samples of Normal control group animal and High fat diet induced animal were sectioned, weighed fixed 10% neutral buffer formalin and mounted in DPX compound. Four micrometer sections were stained with hematoxylin-eosin and PAP stains for the histological examination of micro-vesicular and macro-vesicular steatosis in the liver tissue. The grading of steatosis was given from 0-3 based on the lipid accumulation (steatosis): grade 0 - absence; grade 1 - mild Steatosis (&lt;30% hepatocyte); grade 2- moderate steatosis (30%-70% hepatocyte); grade3- severe steatosis (&gt;70%hepatocytes). Efficacy of grade of PAP stain and H&amp;E stain were accessed and statistically evaluated by considering seven parameters and overall staining characters where compared.&#x0D; Results: Our present study evaluated high fat induced hepatic steatosis of H&amp;E and PAP stain and also evaluated efficacy of both the stain. In our study, Pie chart showing the percentage distribution of Number of micro-vesicles about 70% represents greater than 30, 10% represents 30-70, 20% represents less than 70 (Fig. 1). In our study 20% represents the greater than 30, 30% represents the 30-70, 50% represents Less than 70 (Fig. 2). 30% represents mild steatosis, 20% represents moderate steatosis, 50% represents Severe Steatosis (Fig. 3). &#x0D; Conclusion: We demonstrated the comparative efficiency of the PAP and H&amp;E high fat diet induced rat liver by in vivo and conclude that both H&amp;E and PAP stain has equal efficacy and clarity in observing the various minute parameters.

Gain of Chromosome 6p Correlates with Severe Anaplasia, Cellular Hyperchromasia, and Extraocular Spread of Retinoblastoma.

PurposeGain of chromosome 6p has been associated with poor ocular survival in retinoblastoma and histopathologic grading of anaplasia with increased risk of metastatic spread and death. This study examined the correlation between these factors and other chromosomal abnormalities as well as results of whole genome sequencing, digital morphometry, and progression-free survival.DesignRetrospective cohort study from 2 United States tertiary referral centers.ParticipantsForty-two children who had undergone enucleation for retinoblastoma from January 2000 through December 2017.MethodsStatus of chromosomes 6p, 1q, 9q, and 16q was evaluated with fluorescence in situ hybridization, the degree of anaplasia and presence of histologic high-risk features were assessed by ocular pathologists, digital morphometry was performed on scanned tumor slides, and whole genome sequencing was performed on a subset of tumors. Progression-free survival was defined as absence of distant or local metastases or tumor growth beyond the cut end of the optic nerve.Main Outcome MeasuresCorrelation between each of chromosomal abnormalities, anaplasia, morphometry and sequencing results, and survival.ResultsForty-one of 42 included patients underwent primary enucleation and 1 was treated first with intra-arterial chemotherapy. Seven tumors showed mild anaplasia, 19 showed moderate anaplasia, and 16 showed severe anaplasia. All tumors had gain of 1q, 18 tumors had gain of 6p, 6 tumors had gain of 9q, and 36 tumors had loss of 16q. Tumors with severe anaplasia were significantly more likely to harbor 6p gains than tumors with nonsevere anaplasia (P < 0.001). Further, the hematoxylin staining intensity was significantly greater and that of eosin staining significantly lower in tumors with severe anaplasia (P < 0.05). Neither severe anaplasia (P = 0.10) nor gain of 6p (P = 0.21) correlated with histologic high-risk features, and severe anaplasia did not correlate to RB1, CREBBP, NSD1, or BCOR mutations in a subset of 14 tumors (P > 0.5). Patients with gain of 6p showed significantly shorter progression-free survival (P = 0.03, Wilcoxon test).ConclusionsGain of chromosome 6p emerges as a strong prognostic biomarker in retinoblastoma because it correlates with severe anaplasia, quantifiable changes in tumor cell staining characteristics, and extraocular spread.

Detection of extended-spectrum β-lactamase (ESBL) producing Escherichia coli in chickens

Objective: Indiscriminate use of antibiotics in poultry farms increases the chance of antibiotic-resistant and ESBL producing bacteria in Bangladesh. Therefore, the study was undertaken to detect the extended-spectrum β-lactamase (ESBL) producing Escherichia coli (E. coli) in chickens. &#x0D; Materials and methods: A total of 60 cloacal swab samples (20 from commercial layer, 20 from commercial broiler, and 20 from commercial Sonali chickens) were collected from the Rajshahi district of Bangladesh. The E. coli was isolated from these samples and identified based on cultural, staining, and biochemical characteristics. The disk diffusion method was used to assay the antibiotic-resistant/sensitivity patterns of the isolated E. coli. Phenotypic detection of ESBL producing E. coli was also done. &#x0D; Results: The prevalence of E. coli in chickens was 61.67% in the Rajshahi district of Bangladesh, where the prevalence was 60%, 60%, and 65% in a commercial layer, commercial broiler, and commercial Sonali chickens, respectively. The antibiotic sensitivity assay of E. coli isolated from commercial layer chickens showed 100%, 80%, 50%, 40%, and 40% resistant to amoxicillin, tetracycline, cefotaxime, ciprofloxacin, and ceftazidime, respectively. E. coli isolated from commercial broiler chickens showed 100%, 100%, 60%, 50%, and 40% resistant to amoxicillin, tetracycline, cefotaxime, ceftazidime, and ciprofloxacin, respectively. E. coli isolated from commercial Sonali chickens showed 90%, 70%, 50%, 50%, and 40% resistant to amoxicillin, tetracycline, cefotaxime, ciprofloxacin, and ceftazidime, respectively. In phenotypic detection, the overall prevalence of ESBL producing E. coli was 43.33%, where 40%, 50%, and 40% in the commercial layer, commercial broiler, and commercial Sonali chickens, respectively in the Rajshahi district of Bangladesh. &#x0D; Conclusion: These results indicated that chickens are a potential reservoir for ESBL producing E. coli and their antibiotic resistances are obviously significant. These findings will help us to make a proper guideline for the treatment, prevention, and control of E. coli prevalent in chickens in Bangladesh

Hematologic and Biochemical Reference Intervals and Urinary Test Results for Wild-caught Adult Southern Giant Pouched Rats (Cricetomys ansorgei).

Southern giant pouched rats (Cricetomys ansorgei) are muroid rodents native to subSaharan Africa. They are increasingly used as service animals because of their keen sense of smell and are primarily known for clearing minefields in Africa. The objectives of this study were to determine hematologic and biochemical reference intervals from clinically healthy wild-caught captive adult rats, to describe the cytochemical staining reactions of peripheral blood leukocytes, and to document urinalysis findings. Blood samples were collected from the coccygeal artery of 60 isoflurane-anesthetized rats (36 males and 24 females) and analyzed with automated hematologic and biochemical analyzers; manual differential cell counts were performed on modified Wright-stained blood smears. Urine was collected by cystocentesis, and dipsticks were analyzed on a urine analyzer, with visual examination of unstained sediments. Samples from a male rat with chronic renal disease were excluded from analysis. Reference intervals were determined according to guidelines established by the American Society of Veterinary Clinical Pathology. Lymphocytes were the dominant leukocyte in peripheral blood and granular lymphocytes were identified in most animals. Male rats had significantly higher RBC, absolute reticulocyte counts, and MCV than did female rats. Minor sex-associated differences in urea nitrogen concentration and GGT activity were noted. Leukocytes showed unique cytochemical staining characteristics. Small amounts of protein and bilirubin were found in the urine of rats of both sexes and of female rats, respectively, particularly in concentrated urine. These results will provide benchmarks for determining health status and identifying disease in this species of rat.

Programmed death ligand-1 (PD-L1) immunohistochemical assessment using the QR1 clone in muscle-invasive urothelial carcinomas: a comparison with reference clones 22C3 and SP263.

Programmed death ligand-1 (PD-L1) immunohistochemical (IHC) status is used to predict which patients with metastatic urothelial carcinoma (UC) will respond to immunotherapy. We aimed to compare QR1(Quartett), 22C3 (Dako), and SP263 (Ventana) detection of PD-L1 expression in muscle-invasive UCs and determine the best scoring algorithm for assessment of PD-L1 expression when using the QR1 clone. Our study included 69 UCs. For SP263 and 22C3, PD-L1-positive tumor cell (TC) and/or immune cell (IC) percentages (TC%/IC%) and the Combined Positive Score (CPS) were assessed, respectively (positivity cut-offs of ≥ 25% and ≥ 10). For QR1, both interpretation systems were evaluated. The concordances between assays were calculated. PD-L1 IHC staining characteristics were comparable between QR1, 22C3, and SP263 in both conventional and variant histology UCs. We demonstrated strong or very strong correlations between clones; the strongest correlation for TCs was between QR1 and SP263 (r = 0.92; p = 0.001) and for ICs was between QR1 and 22C3 (r = 0.85; p = 0.001). Our comparative analysis of the scoring algorithms revealed very good concordances among the three assays (range 0.791-0.878); the highest concordance was between QR1 and SP263 when CPS was used as the scoring algorithm for QR1 (0.878; p < 0.001). Our study is the first to demonstrate that the QR1 clone can be used to evaluate PD-L1 status in UCs, with a very good agreement rate with the reference clones. QR1 appeared to be more similar to the SP263 clone. With regard to the scoring algorithm, when evaluating PD-L1 expression using QR1 clone, CPS performed better compared with the TC%/IC% score.

S2623 Chronic Lymphoid Leukemia With Extensive Liver Infiltration in the Absence of Leukemic Transformation

Introduction: Chronic lymphoid leukemia (CLL) is the most common hematologic cancer diagnosed in the elderly. Its association with liver dysfunction secondary to leukemic infiltration ranges from mild elevations in liver enzymes (LE) to acute liver failure. Severe hepatic infiltration is typically associated with acute leukemic transformation. Here we report an atypical manifestation of CLL with rising LE and white blood cell (WBC) count mimicking leukemic transformation. Case Description/Methods: 62-year-old male with history of CLL not on therapy presented to clinic for abnormal LE and rising leukocytosis. Patient reported fatigued, jaundice, right upper quadrant pain and 25lb weight loss over five months attributed to dietary modifications. Patient reported taking glimepiride, atorvastatin and herbal supplements (GNC50+ multivitamin, fertility and prostate multivitamin) that were stopped five months prior to encounter due to concerns of drug-induced liver injury (DILI). Labs were significant for absolute lymphocytosis >53,000/uL, AST 156U/L, ALT 336U/L, total bilirubin 4.4 mg/dl, direct bilirubin 3.57mg/dl and gamma-glutamyl transferase of 94U/L from previously normal LE. Additional work up was negative for viral and autoimmune etiologies. NASH Fibrosure was consistent with F4 Fibrosis. Computer tomography (CT) of abdomen did not demonstrate fatty changes but did show significant abdominal lymphadenopathy. Patient was hospitalized for rapidly rising WBC and LE. Abdominal ultrasound doppler showed patent liver vasculature. Transjugular liver biopsy was negative for acute leukemic transformation, but did identify extensive, >60% sinusoidal infiltration by CLL with hepatic venous pressure gradient (HVPG) of 11 mmHG attributed to periportal and peripancreatic adenopathy observed on CT imaging (Imaging A-D). Patient was initiated on Venetoclax and Obinutuzumab as an outpatient. LE peaked two months after chemotherapy initiation. Liver synthetic function remained normal (Table 1). Discussion: After ruling out DILI with failed improvement upon discontinuation of suspected agents and liver biopsy, extensive CLL extranodal liver involvement was attributed to have caused liver dysfunction and subsequent portal hypertension. Severe pattern of involvement is rare without Richter Transformation, which can be seen in 8% of CLL patients. Due to severe pattern of liver infiltration, worsening LE, portal hypertension, and hepatosplenomegaly it was ultimately decided to start patient on chemotherapy.Figure 1.: Image A:CT abdomen and pelvis with IV contrast depicting an enlarged 33.4x42.9mm periportal lymph node Image B: CT abdomen and pelvis with IV contrast depicting 11.7x18.3mm pancreatic lymph node Image C: : H&E (400x) section demonstrates an atypical lymphoid follicle surrounding bile ductules, with adjacent liver parenchyma. Characteristic immunohistochemical staining of the neoplastic cells shows positivity for CD20 (dim), CD5, CD23, and is negative for CD3 Image D: ZAP 70 (400x) staining (brown) of neoplastic cells has been associated with a poor prognosis.Table 1.: Patient laboratory information from date of diagnosis to most recent outpatient follow up. Footnote: *Represents laboratory data for time of hospital admission for elevated liver enzyme work up **Represents the laboratory data prior to initiation of chemotherapy ***Represents peak liver enzyme abnormality ****Represents most recent outpatient follow up laboratory data.

A Histopathology-based Assessment of Biological Behavior in Oral Hyalinizing Extraosseous Lesions by Differential Stains

The assessment of hyalinization to determine aggressive behavior in oral pathological lesions is a scarcely researched field that requires further exploration. The current study aims to predict the biological behavior of oral hyalinizing extraosseous lesions (OHEOL) by employing four differential stains with clinicopathologic correlation. The study was performed on retrospectively diagnosed formalin-fixed paraffin-embedded cases of salivary gland tumors (SGTs) (n = 13), benign soft tissue (BST) lesions (n = 24), and oral submucous fibrosis (OSMF) (n = 53). The hematoxylin and eosin-stained sections were analyzed for the severity of hyalinization (SOH). Differential stains periodic acid Schiff (PAS), Alcian blue, safranin O, and picrosirius red with polarizing microscopy were used to assess the components of hyalinized tissue. The SOH was correlated with differential staining characteristics and clinicopathologic features to analyze possible correlation with aggressive potential in BST, advancement of disease in OSMF, and recurrence in SGT. Intensity of picrosirius red stain significantly correlated with SOH of SGTs (p = 0.044). The intensity of PAS stain (p = 0.040), picrosirius red polarizing greenish-yellow color (p = 0.002), and pattern of distribution of picrosirius red (p = 0.023) significantly correlated with recurrence of SGTs. The intensity of differential stains increased with the SOH in BST lesions indicating their correlation with SOH. The intensity (p = 0.008) and pattern (p = 0.010) of Alcian blue staining and intensity of safranin O stain (p = 0.003) significantly correlated with SOH in OSMF. Picrosirius red polarizing color reddish and yellowish red (p = 0.002) significantly correlated with SOH distinguishing early and advanced OSMF. Picrosirius red and PAS stains are reliable indicators of SOH and recurrence potential in SGT. Alcian blue, safranin O, and picrosirius red polarizing colors enable detection of SOH and accurately distinguish early from advanced OSMF. SOH can be considered as a histological predictor of aggressive biologic behavior in OHEOL. These findings will result in appropriate management protocols.