Tissue Smears Research Articles

Metastatic cholangiocarcinoma in a cow presenting as suspect bovine tuberculosis in Ireland

AbstractCholangiocarcinoma is a malignant tumour of bile duct epithelial cells, an uncommon primary hepatic neoplasm in cattle. A 9‐year‐old Simmental cow presented with watery diarrhoea, crusty skin lesions, loss of appetite and hepatomegaly. Suspecting severe liver disease, the cow was euthanased due to poor prognosis. During gross postmortem examination, large numbers of multifocal to coalescing nodules often with soft necrotic centres ranging from miliary to 7 cm in size were present in the liver, lung, mesentery, peritoneum and lymph nodes. These features were suggestive of bovine tuberculosis. However, direct tissue smears stained by Ziehl Neelsen stain were negative for the presence of acid‐fast bacteria, and subsequent histopathological and immunohistochemical investigations confirmed the presence of metastasizing cholangiocarcinoma. This case reminds clinicians and pathologists to keep an open mind during the diagnostic process and serves to highlight the value of histopathology in diagnosing seemingly characteristic gross lesions.

Molecular detection of Brucella melitensis in sheep from district Budgam, Jammu and Kashmir

Farmers reported abortions and stillbirths in 600 sheep including 471 pregnant ewes. A complete investigation including physical examination of livestock, postmortem examination of stillbirths, serological examination of ewes and molecular study of abomasal contents, peritoneal fluid, liver and kidney tissue collected from still born lambs was done to ascertain the cause of mortality at Disease Investigation Laboratory, Nowshera, Srinagar. Also, on spot heat fixed tissue smears were prepared for Gram’s staining and accordingly observed for presence of organism. Extraction of bacterial genomic DNA from samples from still born lambs was performed and obtained DNA extracts were checked by agarose gel electrophoresis and stored at -20°C till further use. The PCR conducted on the abomasal, peritoneal contents and liver and kidney tissues revealed 731 bp band after the gel electrophoresis, confirming Brucella melitensis as cause of stillbirths in the flock. The investigation revealed that Brucella melitensis was cause of stillbirths and abortions in sheep flocks.

Developmental stages and molecular phylogeny of Hepatozoon fitzsimonsi (Dias 1953) (Adeleorina: Hepatozoidae) in tortoises Stigmochelys pardalis (Cryptodira: Testudinidae) and ticks of the genus Amblyomma (Acari: Ixodidae) from South Africa

BackgroundHepatozoon fitzsimonsi (Dias, 1953) is a frequently found haemogregarine of southern African tortoises. At the time of this species’ reassignment from the genus Haemogregarina to Hepatozoon, developmental stages such as sporocysts and sporozoites were observed in ticks associated with H. fitzsimonsi parasitised and non-parasitised tortoises. It was thus suggested that ticks may act as the potential vectors for this parasite. However, this earlier research was unable to confirm the identity of these sporogonic stages using molecular markers. In a separate study aimed at identifying tick species parasitising South African reptiles and molecularly screening these for the presence of Hepatozoon, that study identified H. fitzsimonsi in tortoise-associated ticks. Thus, the present study aimed to revisit the potential of ticks to act as vectors for H. fitzsimonsi in tortoises using a combined microscopy and molecular approach.MethodsSpecimens of Kinixys natalensis, Kinixys spekii, Kinixys zombensis and Stigmochelys pardalis were collected from Bonamanzi and Ndumo Game Reserve, South Africa. Upon capture, animals were examined for ticks, and these were collected along with blood and other tissues. Adult ticks were dissected and visceral impression slides were prepared along with thin blood and tissue smears on clean microscope slides. Smears and impression slides were stained with Giemsa, screened and micrographs of parasites were captured. Two primer sets were employed to target fragments of the 18S rRNA gene of parasites found in both tortoises and ticks and the resulting sequences were then compared with other known H. fitzsimonsi and haemogregarine sequences from the GenBank database.ResultsPeripheral blood gamont and liver merogonic stages were observed in S. pardalis, while the sporogonic stages were observed in the haemocoel of Amblyomma ticks. Gamont and sporocyst stages compared morphologically with previous descriptions of H. fitzsimonsi, identifying them as this species. Phylogenetic analysis revealed that the blood and tick sequences obtained in this study clustered in a monophyletic clade comprising known H. fitzsimonsi.ConclusionsThe present study provides further support for ticks acting as the vectors of H. fitzsimonsi by molecularly identifying and linking observed developmental stages in tortoises (S. pardalis) with those in the invertebrate host (Amblyomma spp.).Graphical

Cervical Cancer Detection with a Tissue Smear and a Microscopic Image inside the Deep Learning Model of Squeeze Net

Cervical Cancer Detection with a Tissue Smear and a Microscopic Image inside the Deep Learning Model of Squeeze Net

Impact of transport conditions and underlying disease on post-stocking survival of juvenile Lates calcarifer.

Diseases caused by pathogens commonly occurring in the aquatic environment or those that are non-host specific are prevalent and threaten the rapid growth of tropical aquaculture. This study investigates causes of mortality in 12 batches of newly stocked juvenile Lates calcarifer from three different hatcheries. Cytology based on Diff-Quik™-stained tissue and blood smears provides rapid diagnosis of possible causes of mortality, while histopathology and haematology provide a better understanding of how prolonged transport and fish with existing chronic disease are more likely to experience elevated mortality post-stocking. Our findings showed that accumulation of ammonia during prolonged transport causes extensive damage to epithelial barriers in gastrointestinal tracts and depressed immunity due to marked hypoglycaemia, predisposing fish to acute Streptococcosis. Lates calcarifer with chronic bacterial enteritis developed severe hypoglycaemia, had low circulating total plasma protein, and suffered high mortality within 24 hours post-stocking. Hypoglycaemia and low circulating blood proteins disrupt osmoregulation and exacerbate dehydration, which is fatal in fish in sea water. Dying L. calcarifer tested PCR positive for scale drop disease virus (SDDV) at 28 days post-stocking showed a 10-fold elevation of white blood cell counts, severe vasculitis, and obstruction of blood supply to major organs. Destruction of important immune organs such as spleen is a hallmark of SDDV infection that explains high incidences of opportunistic Vibrio harveyi infections in 61% of fish with SDDV. Overall, this study reiterates the importance of stocking disease-free fish and reducing transport stress.

Leishmania major Infection in Synanthropic Rodents: Evidence for the Urbanization of Zoonotic Cutaneous Leishmaniasis (ZCL) in Southern Iran.

Cutaneous leishmaniasis is of particular importance in southern Iran. This study aimed to investigate the infection of rodents with Leishmania major in an urban area of Fars Province, located in southern Iran. Rodents were trapped and samples from the liver, spleen, and skin were collected. Impression smears were prepared from these tissues and any skin lesions and were examined microscopically. In addition, a portion of the samples were preserved for subsequent DNA extraction. A total of 41 rodents belonging to three species were caught from 10 trapping stations in gardens or houses within the area. The caught rodent species were Rattus rattus (n = 25, 60.97%), Mus musculus (n = 15, 36.58%), and Meriones persicus (n = 1, 2.5%). Leishmania amastigotes were seen in the spleen tissue smear of 6 (2.43%) of the rodents, including 4 of R. rattus and 2 of M. musculus. Skin lesions were observed on the muzzles of two R. rattus and one M. musculus. Samples taken from these lesions tested positive for Leishmania infection. Leishmania DNA was detected in 18 (43.9%) rodents, including 11 R. rattus, 6 M. musculus, and one M. persicus, based on DNA sequencing of the ITS2 gene and PCR of the kDNA. Phylogenetic reconstruction revealed that the parasite infecting the rodents was L. major. The detection of Leishmania infection in these rodents in urban areas raises concerns about the urbanization of cutaneous leishmaniasis caused by L. major. This urbanization poses unique challenges for control and prevention efforts.

Epigenetic variation in neonatal tissues in infants conceived using capacitation-in vitro maturation vs. in vitro fertilization

ObjectiveTo investigate alterations of the global DNA methylation profile in placenta, cord blood and neonatal buccal smears in infants conceived using in vitro maturation (IVM) with a prematuration step (capacitation-IVM [CAPA-IVM] versus in vitro fertilization (IVF). DesignAnalysis of data from the offspring of participants in a randomized controlled trial. SettingPrivate clinic. PatientsForty-six women with polycystic ovary syndrome and/or high antral follicle count and their offspring (58 newborns). Intervention(s)Women with polycystic ovary syndrome and/or a high antral follicle count participating in the clinical trial were randomized to undergo CAPA-IVM or conventional IVF. Main Outcome Measure(s)At delivery, biological samples including cord blood, placental tissue, and neonatal buccal smear were collected. Genome-wide DNA methylation was determined by Illumina Infinium MethylationEPIC BeadChip. Variability in methylation was also considered and mean variances for the two treatment categories were compared. ResultsIn neonatal buccal smears, there were no significant differences between the CAPA-IVM and conventional IVF groups based on CpG probe after linear regression analysis using a significant cut-off of false discovery rate <0.05 and |Δβ|≥0.05. In cord blood, only one CpG site showed a significant gain of methylation in the CAPA-IVM group. In placenta, CAPA-IVM was significantly associated with changes in methylation at five CpG sites. Significantly more variable DNA methylation was found in five probes in placenta, 54 in cord blood and two in buccal smears after IVM of oocytes. In cord blood samples, 20 CpG sites had more variable methylation in the conventional IVF versus IVM group. Isolated CpG sites showing differences in methylation in cord blood were not associated with changes in gene expression of the overlapping genes. Conclusion(s)CAPA-IVM appeared to be associated with only a small amount of epigenetic variation in cord blood, placental tissue and neonate buccal smears.

Molecular identification and infection pathology of Mycobacterium spp. in captive wild animals in Pakistan.

Animal tuberculosis is an infectious, chronic, granulomatous, and debilitating disease affecting animals as well as humans. However, in recent decades, there have been many endemic geographic localities where animal tuberculosis has been identified in wildlife reservoirs, limiting the eradication program in cattle. This study aimed to identify animal tuberculosis in captive zoo animals in Pakistan. In total, 185 morbid zoo animals were brought for postmortem examination at a veterinary postmortem facility. During the macroscopic examination, these animals were thoroughly examined for the presence of suggestive gross lesions of animal tuberculosis (granulomas/tubercles), and the pattern and distribution of these lesions in different organs. The Ziehl-Neelsen (ZN) staining was performed on smears prepared from granulomatous lesions of lung tissue followed by molecular identification of M. bovis and M. tuberculosis DNA using polymerase chain reaction (PCR). The postmortem examination revealed that 8.1% (15/185) of animals had gross tuberculosis lesions on the lungs and lymph nodes. The ZN staining of tissue smears showed 5.40% positivity while M. bovis and M. tuberculosis DNA was identified in 3.78 % and 1.1% of investigated animals, respectively. The study showed that animal tuberculosis is prevalent among wildlife in Pakistan and it may pose serious public health concerns to the people visiting these zoos and wildlife parks.

Rapid detection of Mycobacterium bovis in bovine cytological smears and tissue sections by peptide nucleic acid fluorescence in-situ hybridization

BackgroundBovine tuberculosis is the leading cause of death in cattle and other species worldwide. Quick and precise identification of mycobacteria is critical to control the occurrence of tuberculosis in cattle. MethodsWe developed a fluorescent peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) approach to detect Mycobacterium bovis and Mycobacterium avium in cytological smears and tissue sections of bovines suspected of having tuberculosis. PNA-FISH was conducted on smears of lung and lymph node tissues. Standard bovine mycobacterial cultures were used to standardize the probes using 50 % formamide for M. bovis and 30 % formamide for M. avium. M. bovis probe (MTBCcy3), which was standardized at hybridization conditions of (55 °C and 40 % formamide) concentrations, was positive in all cytological smears. ResultsFour out of twenty five samples tested positive in tissue sections observed as a bright red fluorescence with a cy3 filter (MTBC probe). No results were observed with (MAVTAMRA) probe for M. avium which was standardized at hybridization conditions of (55 °C and 30 % formamide). No fluorescence was observed in the control tissue sections. Additionally, the results were juxtaposed with those of other commonly used detection methods such as immunohistochemistry and Polymerase Chain Reaction (PCR) by targeting the esxA gene. None of the samples tested positive for M. avium infection. ConclusionPNA-FISH can be used to obtain cytological impression smears and tissue sections. When compared to PCR it consumes less time in the diagnosis of bovine tuberculosis in post mortem cases.

Unusual Duodenal Ulcer: Metastatic Urothelial Carcinoma of the Renal Pelvis.

Metastatic urothelial carcinoma of the renal pelvis (MUCP), a type of metastatic upper tract urothelial carcinoma (MUTUC), is a rare malignancy, and some patients with MUCP present with distant metastasis at the time of diagnosis. MUCP in the gastrointestinal tract is even rarer. Herein, we report a 78-year-old man with MUCP that presented as a duodenal ulcer. He complained of anorexia, dizziness, and melena for one month. Endoscopic examination at a local clinic revealed a duodenal hemorrhagic and ulcerative lesion, and the patient was referred. He noted dark-colored stools with increasing frequency, but he denied hematochezia, coffee ground emesis, weight changes, or abdominal pain. Gastroduodenoscopic examination at our hospital demonstrated an ulcerofungating lesion of the second portion of the duodenum. Colonoscopic findings showed no abnormality. Computed tomography showed a 6.7 cm sized mass abutting the inferior vena cava, second portion of the duodenum, lower pole of the right kidney, and right iliopsoas. The mass showed heterogeneous enhancement and internal hemorrhagic necrosis and infiltrated the perinephric soft tissues, the second portion of the duodenum, the right psoas muscle, the right renal vein, and the right adrenal gland. Duodenal biopsy showed moderately differentiated squamous cell carcinoma. Immunohistochemistry (IHC) showed diffuse and strong positivity for CK5/6. Tissue from the liver biopsy showed similar histopathologic features and showed GATA3 positivity on IHC. The imprint cytology smears of the liver tissue showed "cercariform" cell features. We confirmed the diagnosis as MUCP. This case illustrated a rare cause of a secondary duodenal tumor, MUCP.

Touch print smear of testicular tissue with thionine stain for intraoperative diagnosis in azoospermic men.

To compare the diagnostic value of testicular tissue touch print smear (TPS) conducted on azoospermic patients with results obtained from histopathology and in vitro fertility (IVF) lab findings. Microdissection testicular sperm extraction was performed on a group of 148 azoospermic patients and testicular samples obtained intraoperatively. Using TPS, the samples were smeared onto a sterile slide, followed with staining using thionine. The testis tissue bulk samples were also transferred to the IVF lab, and determinations of sperm presence or absence obtained from IVF lab tests were compared with the TPS sample results. Needle testis biopsy was separately performed on a group of 360 azoospermic patients, and results of pathohistology review on the biopsies were further compared with determinations of spermatogenesis stage obtained from TPS for those patients. When compared with IVF lab results, TPS was found to have 100% (126/126) positive predictive value and 95.5% (25/26) negative predictive value for predicting sperm presence or absence, respectively. Furthermore, TPS was further found to have a 93.6% correlation (337 of 360 biopsies) with results of histological diagnoses performed by needle biopsy. Results from histology and TPS for the detection of sperm presence were concordant in 96.1% (346/360) of biopsies. Diagnosis of SCO by TPS shows the highest correlation with histopathology (98.6%), followed by complete spermatogenesis (97.5%), early maturation arrest (78.9%), and late maturation arrest (27.3%). The results support the continued use of TPS in testicular tissue analysis for more rapid assessment of spermatogenesis and for detection of spermatozoa in azoospermic subjects.

PCR for detection of Leishmania donovani from microscopically negative tissue smears of suspected patients in Gondar, Ethiopia.

As untreated visceral leishmaniasis (VL) is fatal, reliable diagnostics are pivotal for accurate treatment allocation. The current diagnostic algorithm for VL in Ethiopia, which is based on the rK39 rapid diagnostic test and microscopy of tissue smears, lacks sensitivity. This probably leads to missed cases and patients not receiving treatment. We conducted a retrospective study on stored microscopically negative spleen and bone marrow smears from suspected VL patients collected at the Leishmaniasis Research and Treatment Center (LRTC) in Gondar, northern Ethiopia between June 2019 and November 2020. Sociodemographic, clinical and treatment data were collected and samples were tested by real-time PCR targeting kinetoplast DNA. Among the 191 eligible samples (135 spleen and 56 bone marrow) with a microscopically negative and valid PCR result, 119 (62.3%) were positive by PCR, although Ct values for some were high (median 33.0). Approximately three quarters of these undiagnosed primary VL (77.3%) and relapse (69.6%) patients did not receive antileishmanial treatment. Of the 56 microscopically negative bone marrow samples, 46 (82.1%) were PCR positive, which is considerably higher compared to the microscopically negative spleen samples, for which 73 out of 135 (54.1%) were PCR positive. The odds of being PCR positive were significantly higher for bone marrow aspirates and higher when white blood cell values were lower and splenomegaly (in cm) was more pronounced. This study demonstrates that a lot of suspected VL patients remain undiagnosed and untreated. This indicates the urgent need for better diagnostics for VL in the East-African region. The outcomes of PCR positive should be closely monitored and treatment should be provided if the patient deteriorates. In resource limited settings, implementation of PCR on bone marrow aspirate smears of patients with low WBC values and splenomegaly could lead to considerable improvements in patient management.

Parasitological, Molecular, and Histopathological Investigation of the Potential Activity of Propolis and Wheat Germ Oil against Acute Toxoplasmosis in Mice

Toxoplasmosis is one of the most common parasitic zoonoses that affects all vertebrates. The drugs most commonly used against toxoplasmosis have many side effects, making the development of new antiparasitic drugs a big challenge. The present study evaluated the therapeutic effectiveness of novel herbal treatments, including propolis and wheat germ oil (WGO), against acute toxoplasmosis. A total of 50 albino mice were divided into five groups: group 1 (G1) (non-infected and non-treated); group 2 (G2) (infected without treatment); group 3 (G3) (treated with propolis); group 4 (G4) (treated with WGO); group 5 (G5) (treated with a combination of propolis and WGO). The effects of the herbal substances on different organs, mainly liver, spleen, and lungs, were investigated using parasitological, molecular, and histopathological examinations. The results of parasitological examination demonstrated statistically significant (p < 0.05) differences in the parasitic load between treated groups (G3, G4, and G5) compared to the control positive group (G2). These differences were represented by a significant reduction in the parasite load in stained tissue smears from the liver obtained from the animals treated with propolis (G3) compared to the parasite load in the positive control group. Similarly, animals (G4) treated with WGO exhibited a significant reduction in the parasite load versus the positive control group, while the lowest parasite load was found in G5, treated with propolis and WGO. Quantification of the parasite burden through molecular methods (PCR) revealed similar findings represented by reduction in the parasite burden in all treated groups with WGO and propolis as compared to the control group. Importantly, these previous parasitological and molecular findings were accompanied by a marked improvement in the histopathological picture of the liver, spleen, and lungs. In conclusion, propolis and WGO showed a good combination of therapeutic efficacy against acute toxoplasmosis.

Granuloma inguinale diagnosed on the bedside.

Donovanosis is a chronic bacterial infection caused by gram-negative bacterium Klebsiella granulomatosis and is characterized by painless beefy red granulomatous ulcer which bleeds on touch and is mostly confined to the endemic regions of the world. It is rarest among the sexually transmitted genital ulcer diseases. We hereby report a case of donovanosis in a non-endemic area who initially presented with a solitary genital ulcer. Initial tests did not reveal any aetiology. However after four days of admission, since no aetiology was coming forth, a repeat tissue smear from the lesion revealed Donovan bodies which helped in the bedside diagnosis and management of the case.

Fourier Imager Network (FIN): A deep neural network for hologram reconstruction with superior external generalization

Deep learning-based image reconstruction methods have achieved remarkable success in phase recovery and holographic imaging. However, the generalization of their image reconstruction performance to new types of samples never seen by the network remains a challenge. Here we introduce a deep learning framework, termed Fourier Imager Network (FIN), that can perform end-to-end phase recovery and image reconstruction from raw holograms of new types of samples, exhibiting unprecedented success in external generalization. FIN architecture is based on spatial Fourier transform modules that process the spatial frequencies of its inputs using learnable filters and a global receptive field. Compared with existing convolutional deep neural networks used for hologram reconstruction, FIN exhibits superior generalization to new types of samples, while also being much faster in its image inference speed, completing the hologram reconstruction task in ~0.04 s per 1 mm2 of the sample area. We experimentally validated the performance of FIN by training it using human lung tissue samples and blindly testing it on human prostate, salivary gland tissue and Pap smear samples, proving its superior external generalization and image reconstruction speed. Beyond holographic microscopy and quantitative phase imaging, FIN and the underlying neural network architecture might open up various new opportunities to design broadly generalizable deep learning models in computational imaging and machine vision fields.

Subcutaneous Benign Tumour; A Case of an Ox and Its Management Outcome; A Case Report

Tumor or neoplasm is an abnormal mass or swelling of tissue which results when cells divide more than they should beyond brain’s control. This case report describes, a successful management of subcutaneous benign tumour on an adult local breed ox having a body weight of 295 kg that was presented to VTH of AAU-CVMA, Bishoftu, on February 24/2022. It has a primary complaint and a history of firm and cold swelling at the caudal part of mandible on the lateral side of neck region that has spent over four years. Aspiration of the swelling with a sterile 16 gauge needle with syringe didn’t detect or withdrawn any of the contents (pus, fluid, blood and other). Cytological examination of tissue smears resulted immature and irregular shaped cells which have large darker nucleus. Based on history, clinical signs and cytological staining result, the case was diagnosed as subcutaneous benign tumor which is differentially diagnosed from abscess, oedema, cascious lymph adenitis and haematoma. It was managed by surgical resection and intramuscular injection of penicillin G procaine with wound spray topically for three successive days. Complete recovery of the ox was achieved after three weeks. Surgical resection accompanied with topical application and parenteral administration of antibiotics can give successful outcome on the management of localized subcutaneous benign tumours after proper identification and effective analgesics.

Comparison among Four Deep Learning Image Classification Algorithms in AI-based Diatom Test.

To select four algorithms with relatively balanced complexity and accuracy among deep learning image classification algorithms for automatic diatom recognition, and to explore the most suitable classification algorithm for diatom recognition to provide data reference for automatic diatom testing research in forensic medicine. The "diatom" and "background" small sample size data set (20 000 images) of digestive fluid smear of corpse lung tissue in water were built to train, validate and test four convolutional neural network (CNN) models, including VGG16, ResNet50, InceptionV3 and Inception-ResNet-V2. The receiver operating characteristic curve (ROC) of subjects and confusion matrixes were drawn, recall rate, precision rate, specificity, accuracy rate and F1 score were calculated, and the performance of each model was systematically evaluated. The InceptionV3 model achieved much better results than the other three models with a balanced recall rate of 89.80%, a precision rate of 92.58%. The VGG16 and Inception-ResNet-V2 had similar diatom recognition performance. Although the performance of diatom recall and precision detection could not be balanced, the recognition ability was acceptable. ResNet50 had the lowest diatom recognition performance, with a recall rate of 55.35%. In terms of feature extraction, the four models all extracted the features of diatom and background and mainly focused on diatom region as the main identification basis. Including the Inception-dependent model, which has stronger directivity and targeting in feature extraction of diatom. The InceptionV3 achieved the best performance on diatom identification and feature extraction compared to the other three models. The InceptionV3 is more suitable for daily forensic diatom examination.

Evaluation of antimicrobial action, pulp tissue dissolution, and smear layer removal of an experimental irrigant combining Salvadora persica and sodium hypochlorite with and without ultrasonic activation: An in vitro study

Aim: To evaluate the pulp tissue dissolution, antimicrobial action, and smear layer removal using an experimental irrigant combining Salvadora persica and sodium hypochlorite (NaOCl) with and without ultrasonic activation in comparison to standard irrigation protocol of NaOCl and ethylenediamine tetraacetic acid (EDTA) with and without ultrasonic activation.Materials and Methods: Pulp tissue dissolution - Pulp tissue weighing 0.9 g was dissolved in each experimental Group G1: 5 ml of 5% NaOCl, G2: G1 + ultrasonic activation, G3: 5 ml 5% NaOCl + 40% S. persica, G4: G3 + ultrasonic activation and weighed after 1, 5, and 10 min. The difference in weight of the tissue sample was assessed. Smear layer removal and calcium/phosphate (Ca/PO4) ratio - G1: 5 ml of 5% NaOCl, saline, 1 ml of 17% EDTA, G2: G1 + ultrasonic activation, saline, 17% EDTA, G3: 5 ml 5% NaOCl + 40% S. persica G4: G3 + ultrasonic activation. Each tooth was then split longitudinally and subjected to scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM-EDX) analysis. Antibacterial Activity- assessed against Enterococcus Faecalis.Statistical Analysis: SEM, pulp tissue dissolution and antimicrobial efficacy – Kruskal–Wallis and post hoc Mann–Whitney EDX – ANOVA and Tukey's honestly significant difference post hoc analysis.Results: Pulp dissolution by G3 and G4 was better. However, there was no statistically significant difference between the groups (P > 0.05). Experimental combination performed better in terms of smear layer removal, showed higher amount Ca/PO4 in EDX analysis, but the difference was not statistically significant between the groups (P > 0.05). G2 showed maximum antimicrobial efficacy compared to all groups but not statistically significant (P > 0.05).Conclusions: The combined formulation of 5% NaOCl + 40% S. Persica was as effective as conventional protocol and ultrasonic activation improved its efficacy.

Determination of microsporidia infection in larvae and adult anopheles mosquitoes (Diptera: Culicidae) Northwest Ethiopia

Microsporidia are a diverse group of obligate intracellular, spore-forming parasites that infect invertebrates including mosquitoes. A few studies showed that microsporidia infection of mosquitoes is associated with larval mortality or reduced adult fecundity and lifespan. This phenomenon might be used to protect against vector-borne disease transmission. This study aimed to determine microsporidia infection in field-collected Anopheles larvae and reared adults. Anopheles larvae were collected from the field and analyzed from February to April 2022. After determining the species identity of the 4th stage larvae, they were dissected to extract their midgut while the early-stage larvae were transported to an entomological laboratory for rearing the adult in the insectary room. Similarly, after determining the species identity of adult female Anopheles mosquitoes, they were dissected to extract their midgut. Then, two thin smears for each dissected larvae and/ adults were made. Fixed preparations were stained with Giemsa and modified Ziehl-Neelsen staining techniques to detect the microsporidia. Descriptive statistics and an independent t-test were used to analyze the data. Anopheles gambiae complex was identified as the most predominant species from field-collected larvae and insectary reared adults. From the total 258 larvae and 258 adults An.gambiae complexes tissue smears examined, microsporidia were detected in 2.7% [7/258, (95%, CI: 0.8-5)] and 1.2% [3/258, (95%, CI: 0.01-2.7)] of larvae and adults respectively. The mean density of microsporidia in larvae and adults was 64.9 (± 23.4 SD) and 36 (± 8.5SD) respectively. The difference in microsporidia density between infected larvae and adults was statistically significant (F=1.77, P= 0.02). Conclusion: Significant level of microsporidia infection was detected using light microscopy. Further microsporidia identification in the genus and species level is needed.

Intraoperative Assessment of IDH Mutation Status and Tumor Invasioni in Glioma

Abstract Introduction/Objective Maximizing surgical resection in gliomas, while avoiding compromising non-infiltrated tissue, is associated with survival benefit. Current methodologies are suboptimal in providing rapid, intraoperative molecular characterization of tissue. We address this unmet need by using desorption electrospray ionization mass spectrometry (DESI-MS) for the intraoperative molecular assessment of gliomas. Methods/Case Report This prospective study uses intraoperative DESI-MS analysis of fresh tissue to evaluate IDH mutations via 2-hydroxyglutarate intensity and TCP via measurement of N-acetylaspartic acid (NAA) intensity and characteristic lipid profiles in less than three minutes. Blinded review of the tissue smears by a neuropathologist is used to validate IDH mutation status and TCP estimates. Results (if a Case Study enter NA) Presently, 529 biopsies from 85 enrolled patients have been collected and analyzed at two institutions. TCP assessment based on NAA intensity in 203 biopsies at the first institution yielded sensitivity, specificity, and accuracy values of 91, 76, and 83%, whereas TCP estimates via characteristic lipid profiles yielded 76, 85, and 81%, respectively. Assessment of IDH mutation status of 71 core biopsies yielded sensitivity, specificity, and accuracy values of 89, 100, and 94%. Ongoing validation of the methodology is being performed at a second institution, where we have collected 282 biopsies from 36 patients. IDH mutation assessment of the first 15 patients indicate 100% sensitivity, specificity, and accuracy. Conclusion This study represents the first and largest study using DESI-MS for the intraoperative evaluation of IDH status and TCP measurement in gliomas. Prospectively, we propose to modify our DESI-MS system to allow estimation of IDH mutation status and TCP in surgical cavities without the need for a biopsy by placing a surgical material along the margin and transferring material from the blot to a microscope slide prior to DESI-MS analysis. We envision molecular analysis by DESI-MS as a complementary technique to histopathology capable of providing additional clinical information in near real-time.