Histopathological Methods Research Articles

A Histopathological Image Classification Method Based on Model Fusion in the Weight Space

Automatic classification of histopathological images plays an important role in computer-aided diagnosis systems. The automatic classification model of histopathological images based on deep neural networks has received widespread attention. However, the performance of deep models is affected by many factors, such as training hyperparameters, model structure, dataset quality, and training cost. In order to reduce the impact of the above factors on model training and reduce the training and inference costs of the model, we propose a novel method based on model fusion in the weight space, which is inspired by stochastic weight averaging and model soup. We use the cyclical learning rate (CLR) strategy to fine-tune the ingredient models and propose a ranking strategy based on accuracy and diversity for candidate model selection. Compared to the single model, the weight fusion of ingredient models can obtain a model whose performance is closer to the expected value of the error basin, which may improve the generalization ability of the model. Compared to the ensemble model with n base models, the testing cost of the proposed model is theoretically 1/n of that of the ensemble model. Experimental results on two histopathological image datasets show the effectiveness of the proposed model in comparison to baseline ones, including ResNet, VGG, DenseNet, and their ensemble versions.

Effects of destruxin A on hemocytes of the domestic silkworm, Bombyx mori.

Destruxin A (DA) is a mycotoxin isolated from the entomopathogenic fungus Metarhizium anisopliae which has demonstrated inhibitory activity against various insect species. However, the mechanism of inhibition on target sites in insects remains unknown. In this research, the dose-response relationship between DA and morphological changes in body tissues and organs of domestic silkworm, Bombyx mori, were investigated by histopathological methods to identify the target sites that responded to DA. The results showed that responses of individual tissues and organs varied with DA dosage and treatment time. At low doses (i.e., 0.01μg/g), the hemocytes were the most sensitive to DA with morphological changes apparent at 6 h after treatment. However, the muscle cells, fat body, and Malpighian tubules were unaltered. At higher doses (i.e., > 0.1μg/g), morphological changes were observed in muscle cells, fat body, and Malpighian tubules at 24 h after treatment. The results indicated that DA can be an immunosuppressant by damaging host cells like hemocytes, and at higher doses may potentially impact other physiological processes, including muscle function, metabolism, and excretion. The information presented in the current study will facilitate development of mycopesticides and novel immunosuppressants.

PREDICTION OF COLLAGEN CONTENT THROUGH BIOMECHANICAL PARAMETERS IN MICE SKIN WOUND: A COMPARISON OF ANN AND ANFIS MODELS

Pathological analysis as well as biomechanical methods are powerful approaches for collagen assessment, which plays an important role in understanding the wound healing process and choosing a treatment method in clinical situations. Due to the limitations of preparing and evaluating pathological images, this study was designed to establish a machine learning technique to predict the wound collagen content through its biomechanical parameters. For this purpose, the artificial neural network (ANN) and adaptive neuro-fuzzy inference system (ANFIS) were compared. The wound was created with an incision on the back of 30 male BALB/c mice. On the 7th and 14th days, animals were sacrificed and 60 wound tissue samples were evaluated using histopathological and biomechanical methods to quantify the amount of collagen and wound tensile strength to feed the ANN and ANFIS developed models. Based on the results, both models have appropriate performance to predict the wound collagen content. However, the comparison of coefficient of determination ([Formula: see text]) and root mean square error (RMSE) for testing dataset revealed that ANN ([Formula: see text], [Formula: see text]) had more prediction capability than ANFIS ([Formula: see text], [Formula: see text]). As a decision support system, ANN model could assist in the evaluation of wound healing process with collagen values prediction.

The intervention effect of Aitongxiao prescription on primary liver cancer rats was evaluated based on high-throughput miRNA sequencing and bioinformatics analysis.

Liver cancer is a common malignant tumor known for its difficult treatment and poor prognosis. As a traditional Chinese medicine prescription, Aitongxiao prescription (ATXP) has been used in clinical treatment of primary liver cancer (PLC) for more than ten years, and its therapeutic effect is obvious and has been verified over time. However, the mechanism of ATXP in treating PLC has not been fully elucidated. This study aimed to detect the liver-protective effect of ATXP on a PLC rat model and explore its potential mechanism from the perspective of plasma extracellular vesicle miRNAs. Fifty SPF male SD rats were randomly selected, with six rats as the control group, and the remaining rats were injected with DEN to establish a primary liver cancer model. The model rats were randomly divided into the model group and the ATXP group. After 4 weeks of intervention, the liver-protective effect of ATXP was evaluated using plasma biochemical indicators and histopathological methods. Plasma extracellular vesicles were isolated and extracted, and identified by transmission electron microscopy, nanoparticle tracking analysis, and western blot. Significant differentially expressed miRNAs in extracellular vesicles were screened by Illumina sequencing to explore the therapeutic targets of ATXP and conduct functional analysis. The results showed that ATXP significantly reduced plasma liver function in PLC rats and alleviated liver pathological damage. In addition, plasma extracellular vesicles were isolated and identified. According to the results of GO and KEGG analysis, they were related to multiple biological processes and covered multiple signaling pathways (PI3K-Akt and MAPK signaling pathways, etc.). The interaction between miR-199a-3p and MAP3K4 was determined by bioinformatics methods and dual-luciferase reporter gene detection, confirming that MAP3K4 is the target gene of miR-199a-3p. In conclusion, ATXP protects the liver from DEN-induced PLC, which may be related to the regulation of plasma extracellular vesicle miR-199a-3p. This study further reveals the mechanism of ATXP in treating liver cancer and provides a theoretical basis for subsequent research.

Anthocyanins improve liver fibrosis in mice by regulating the autophagic flux level of hepatic stellate cells by mmu_circ_0000623.

Liver fibrosis is a key step in the progression of various chronic liver diseases to liver cirrhosis and even liver cancer, it is also an important link affecting prognosis. Therefore, this study aimed to investigate the therapeutic effect of anthocyanins on liver fibrosis and the molecular mechanism of mmu_circ_0000623 in anthocyanin therapy. In this study, CCL4 was used to build a mouse liver fibrosis model, and the treatment groups were treated with 100 and 200 mg/kg of anthocyanins daily by gavage. Liver fibrosis indicators, macrophage polarization markers, and liver injury markers were further detected by real-time quantitative PCR (qRT-PCR), western blotting (WB), and enzyme-linked immunosorbent assay. Morphological verification of liver injury in different treatment groups was performed by histopathological method. A mouse hepatic stellate cell (HSC) model and a mouse liver fibrosis model were constructed to verify the expression of circ_0000623, miR-351-5p, and TFEB. Transfected with mRFP-GFP-LC3 to detect the autophagic flux of HSCs. We found that 100 mg/kg or 200 mg/kg of anthocyanins could significantly reduce the degree of liver fibrosis in mice. In addition, anthocyanins can inhibit the proliferation, activation, and migration ability of HSCs. circ_0000623 was lowly expressed in mice with liver fibrosis, and anthocyanin treatment could promote its increased expression. Further testing found that anthocyanins could reverse the blocked autophagic flux induced by PDGF or CCL4. This effect is achieved by regulating the expression of TFEB by competitive adsorption of miR-351-5p. Anthocyanins could treat liver fibrosis by modulating circ_0000623/miR-351-5p/TFEB-mediated changes in HSC autophagic flux.

CORRELATION OF PLASMA D-DIMER LEVEL IN FEMALE BREAST CARCINOMA PATIENTS WITH CLINICAL STAGES AND HISTOPATHOLOGICAL PARAMETERS

Background: Breast cancer is one of the leading disease among females in the world. There is a correlation between cancer and hypercoagulation. In a cancer state, there is an increase in the level of cross-linked brin degradation product (d-dimer) which indicates systemic activation of brinolysis and hemostasis. So, there is a relation between an increase in d-dimer value and breast carcinoma.Objective:This study was done in an attempt to evaluate the role of D-dimer in patients with carcinoma breast, in predicting lymph node metastasis in carcinoma patients and to look for the relationship of these markers with histopathologic parameters Method: A prospective observational descriptive study was done in Dr. S N medical college attached hospitals (department of surgery) from Jan 2022 to Jan 2023. Forty-three breast carcinoma patients were included in the study after core needle biopsy conrmation. Plasma d-dimer levels compared with (tumor size, stage, grade, lymphovascular invasion, and lymph node involvement). Data collected and analysed. Results: Forty three patients were studied with age from 21 to 76yrs(mean age-51).left side tumor was more common 52%. D-dimer level was increasing with enlarged tumor size, high stage, high grade, lymphovascular invasion, and lymph node involvement. All are statstacally signicant.(p value: <0.001) Conclusions: Plasma D-dimer level is important preoperative marker with which we can predict the advance stage breast cancer. It is also an important marker of clinical stage progression, lymphovascular invasion, and metastasis . Pre-operative plasma D-dimer level is a safe, cost-effective, and convenient method for the prediction of advanced-stage breast cancer

Evaluation of mono and combined nitrofurantoin therapy for toxoplasmosis in vivo using murine model

ABSTRACT Toxoplasmosis is a frequent disease with an estimated prevalence of more than one billion human cases worldwide and over one million new infections each year. It is classified as a neglected tropical disease by the CDC since 2019. The disease may pass unnoticed in healthy individuals but could be fatal in the immunocompromised. Moreover, no effective treatment is available against the chronic form of the disease. Available anti-Toxoplasma drugs are associated with many side effects. Therefore, search for new more reliable, more efficient, and less toxic therapeutic agents is a continuous endeavor. This study assesses the potential use of nitrofurantoin, a compound with well-established antimicrobial properties, as a potential anti-Toxoplasma drug in vivo. It compares its efficacy to the commonly used anti-Toxoplasma agent spiramycin by molecular and histopathological methods in acute and chronic infection. The results demonstrate a significant ability to eliminate the parasite (P < 0.001) whether used as mono- or combined therapy with spiramycin in the acute and chronic stages. When compared to the anti-Toxoplasma drug spiramycin, nitrofurantoin achieved similar efficacy in the acute and chronic infection (P = 0.65 and P = 0.096, respectively). However, better results were obtained when using a combination of both drugs (P < 0.001). Additionally, nitrofurantoin showed good inhibitory effects on the inflammatory process in the liver, kidney, and uterus of the experimentally infected animals. In conclusion, nitrofurantoin can be considered as a potential anti-Toxoplasma agent. Nevertheless, further studies are recommended before consideration for clinical trials.

Two flavonoids, baicalein and naringin, are effective as anti-inflammatory and anti-oxidant agents in a rat model of polymicrobial sepsis

Introduction In this study, our aim was to investigate the possible protective and therapeutic effects of these two flavonoids, baicalein, and naringin, in 50 and 100 mg/kg doses applied both before and after cecal ligation and puncture (CLP) procedures in a polymicrobial sepsis rat model, and evaluate the possible contribution of oxidative and inflammatory markers by immunological, biochemical, molecular, and histopathological methods. Methods Sixty-six Wistar albino rats were divided into 11 groups. The pro-inflammatory (TNF-alpha, IL-1-beta, and IL-6) and anti-inflammatory (TGF-beta and IL-10) cytokine levels were measured by ELISA technique. CD3, CD68, and nuclear factor kappa B positivity rates were detected by immunohistochemical methods. Oxidative stress parameters (MDA, SOD, and GSH) were measured by tissue biochemistry. Results Sepsis caused a significant increase in all pro-inflammatory cytokine levels and MDA activity. Also, it led to an increase in the positivities of CD3, CD68, and NF-κB markers. However, especially pre-CLP doses of baicalein and naringin inhibited the inflammation process by suppressing pro-inflammatory and increasing anti-inflammatory cytokine levels, as well as regulating the oxidative stress process by normalizing the oxidant/anti-oxidant enzyme levels. Conclusion Both pre- and post-application of baicalein and naringin are quite effective to prevent sepsis-caused cellular processes. This protective and therapeutic effects by baicalein and naringin in animals with sepsis seems to be originated from the high antioxidant capacity and inhibition of pro-inflammatory cytokine production. Thus, those natural agents may prove to be valuable protective agent against septic shock.

Quality Assurance and Cost Reduction in Histopathology Laboratories Using Tissue Microarrays

In the context of cost increases of both labor and consumables, cheaper and faster histopathology methods are needed. We implemented in our research laboratory the use of tissue microarrays (TMA) for the parallel processing and analysis of tissue samples. In this study, we used seven pre-processed, paraffinated biomimetic sectionable support matrices serving as "recipient" paraffin blocks to embed a total of 196 tissue cores from formalin-fixed paraffin-embedded tissue samples (serving as "donor" paraffin blocks) from seven different rabbit organs. These tissue samples were obtained using four different processing protocols: two 6 h protocols with xylene as the transition solvent, and two using butanol instead (one 10 h in duration and the other 72 h long). While the samples from protocols 1 and 2 (with xylene) quite regularly generated peeling of some of the cores from the slides (most likely because of substandard paraffin infiltration), butanol processing performed flawlessly for both processing protocols. Our proposed technique of using TMAs in the research laboratory brings with it a significant reduction in time and consumable costs (up to 77 and 64%, respectively), but also new challenges for all the upstream processes.

Abstract 2397: An advanced approach technique for tracking point between immune check point inhibitor and cancer cell mechanism in immune response motility by infiltration a breast cancer cell

Abstract In Breast Cancer, Tumor-infiltrating lymphocyte (TIL) motility has been reported that it is reflecting tumor microenvironment response by adoptive cell therapy (ACT). ACT is well-known to be best applied for suppressing cancer cell growth with immune response and it also was well-known the immunotherapy. However, ACT response tracking by general analysis method included immunohistochemistry and flow cytometry was provided limited information for cancer therapy in recent years. Therefore, by assessing relationship immunotherapy marker, PD-L1, known as immune check point inhibitor and TIL using intravital microscopy in breast cancer model, we also imaged whether TIL response was related to immunotherapy following dependent on time point in a various breast cancer cell implanted mouse model such as 4T1 (mouse breast cancer cell), MDA-MB-231 (human triple negative breast cancer cell) and MCF7 (human breast cancer cell). These images result in PD-L1 expression was decreased in cancer cell implanted until 10days, whereas TIL expression increased in a various breast cancer cell when it is measuring displacement length, track mean velocity and confinement ratio called tumor microenvironment. These results indicated that we can imaged immune response related to TIL response in breast cancer cell line by intravital microscopy instead of general microscopy. It means that intravital microscope was an advanced approach technique to understand cancer mechanism accompanied by the immune response and established the method of high-resolution intravital cellular visualization compared to a general histopathological method for evaluating immune response in cancer tissue Citation Format: Hyunseok Kim, Soyeon Ahn, Hyungjin Kwon, Hsuan-Ju Chen, Inwon Park, Pilhan Kim, Akyildiz Kubra. An advanced approach technique for tracking point between immune check point inhibitor and cancer cell mechanism in immune response motility by infiltration a breast cancer cell [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2397.

Lin28 promoting the protective effect of PMSCs on hepatic ischaemia-reperfusion injury by regulating glucose metabolism.

Human placental mesenchymal stem cells (PMSCs) can prevent liver ischaemia-reperfusion injury (LIRI). However, their therapeutic effects are limited. Therefore, additional research is required to elucidate the mechanisms of PMSC-mediated LIRI prevention and enhance the related therapeutic effects. This study aimed to examine the role of the Lin28 protein in the regulation of glucose metabolism in PMSCs. Further, it explored whether Lin28 could enhance the protective effects of PMSCs against LIRI and investigated the underlying mechanisms. Western blotting was performed to examine Lin28 expression in PMSCs under hypoxic conditions. A Lin28 overexpression construct was introduced into PMSCs, and the effect on glucose metabolism was examined using a glucose metabolism kit. Further, the expression of some proteins involved in glucose metabolism and the PI3K-AKT pathway and the levels of microRNA Let-7a-g were examined using western blots and real-time quantitative PCR, respectively. To examine the relationship between Lin28 and the PI3K-Akt pathway, the effects of AKT inhibitor treatment on the changes induced by Lin28 overexpression were examined. Subsequently, AML12 cells were co-cultured with PMSCs to elucidate the mechanisms via which PMSCs prevent hypoxic injury in liver cells in vitro. Finally, C57BL/6J mice were used to establish a partial warm ischaemia-reperfusion model. The mice received intravenous injections containing PMSCs (control and Lin28-overexpressing PMSCs). Finally, their serum transaminase levels and degree of liver injury were assessed using biochemical and histopathological methods, respectively. Lin28 was upregulated under hypoxic conditions in PMSCs. Lin28 exerted protective effects against hypoxia-induced cell proliferation. Moreover, it increased the glycolytic capacity of PMSCs, allowing PMSCs to produce more energy under hypoxic conditions. Lin28 also activated the PI3K-Akt signalling pathway under hypoxic conditions, and its effects were attenuated by AKT inhibition. Lin28 overexpression could protect cells against LIRI-induced liver damage, inflammation and apoptosis and could also attenuate hypoxia-induced hepatocyte injury. Lin28 enhances glucose metabolism under hypoxic conditions in PMSCs, thereby exerting protective effects against LIRI by activating the PI3K-Akt signalling pathway. Our study is the first to report the potential of genetically modified PMSCs for LIRI treatment.

Utility of clinicoradiological, microbiological, histopathological, and molecular methods in the diagnosis of spinal tuberculosis.

The diagnosis of STB is mainly based on clinicoradiological observations substantiated by bacterial culture, staining, Gene Xpert, and histopathology. The purpose of the study was to correlate these methods to evaluate the effectiveness in the diagnosis of STB. A total of 178 clinicoradiologically suspected cases of STB were included in the study. The specimens for diagnostic workup were collected either during surgery or by CT-guided biopsy. All these specimens were tested for tuberculosis through ZN staining, solid culture, histopathology, and PCR. The sensitivity, specificity, PPV, and NPV of each test were calculated using histopathology as a gold standard. Out of the 178 cases, a total of 15 cases were excluded from this study. Among the remaining 163 cases, TB was diagnosed in 143 [87.73%] on histopathology, 130 [79.75%] on Gene Xpert, 40 [24.53%] on culture, and 23 [14.11%] on ZN stain. The sensitivity, specificity, PPV, and NPV of Gene Xpert were 86.71, 70, 95.38, and 42.42%, respectively. The sensitivity, specificity, PPV, and NPV of AFB culture were 27.97, 100, 100, and 16.26%, respectively. The sensitivity, specificity, PPV, and NPV of AFB stain were 16.08, 100, 100, and 14.29%, respectively. Gene Xpert showed a moderate agreement [Ƙc = 0.4432] with histopathology. No single diagnostic modality can ascertain the diagnosis, and it is desirable to have a combination of diagnostic batteries for better results. A combination of Gene Xpert and histopathology aids in early and reliable diagnosis of STB.

456 Analysis of Clinical, Histologic, and Molecular Characteristics of Proliferative Verrucous Leukoplakia

OBJECTIVES/GOALS: This study aims to develop objectively scored histological characteristics of early oral leukoplakia, which may be correlated with molecular pathways predictive of progression into proliferative verrucous leukoplakia (PVL). The secondary aim is to develop a biomarker profile to be used in diagnosis, staging, and management of PVL. METHODS/STUDY POPULATION: Clinical and pathology records of 120 patients with oral leukoplakia and/or PVL were reviewed. Eight patients were selected—all had serial biopsies over time leading to PVL suspicion. Specimens were deidentified and subjected to blinded examination by a board certified oral pathologist, then scored relative to the extent of each of the commonly accepted histologic characteristics of PVL: hyperkeratosis, acanthosis, blunt rete ridges, hyperchromatic nuclei, increased nuclear-cytoplasmic ratio, dyskeratosis, and surface corrugation. Given these results, a larger subset of patient samples will be labeled and assayed for expression of epidermal growth factor receptor tyrosine kinases and downstream pro-oncogenic signaling mediators. Expression of these factors will be tested against progression to PVL. RESULTS/ANTICIPATED RESULTS: Histologically, in scoring the specimens from eight subjects, the characteristics of acanthosis, dyskeratosis, and blunted rete ridges had the strongest correlation with eventual progression to PVL. These criteria will therefore be recommended as an objective histopathologic method of identification of patients with high risk of development of PVL, and therefore malignant potential. We expect the results of the biomarker assay to provide a molecular basis for predicting PVL pathogenesis. Particularly, we anticipate pro-oncogenic targets such as EGFR, PI3K, Akt, and mTOR pathways will show increased expression as leukoplakic lesions progress. These results would then provide the basis for testing patient samples for expression of these markers in a longitudinal study of PVL emergence and progression. DISCUSSION/SIGNIFICANCE: The aggressive nature of PVL, with a rate of malignant transformation of 61% and mortality rate of 40%, requires close clinical monitoring in order to improve patient outcomes. Therefore, well defined objective clinical, histologic, and molecular criteria are critical for early detection of sites likely to progress to PVL and subsequent malignancy.

Histopathological and molecular methods as complementary diagnostic in case of lymphadenopathies suggestive of bovine tuberculosis.

Bovine tuberculosis (TB) is a chronically evolving zoonotic infectious disease caused by Mycobacterium bovis. Anatomopathological examination during post mortem inspection in bovines is the main resource engaged in sanitary slaughter; however, it is very troublesome since many granulomatous inflammatory processes have similar morphological characteristics. Thus, this study aims to use complementary diagnosis methods (histopathological and polymerase chain reaction - PCR assays) to confirm the macroscopic assessment of lymphadenopathies indicative of tuberculosis in bovines slaughtered in a refrigerated slaughterhouse in Tailândia city, PA, Brazil. Fifty‑one samples were collected from lesions indicative of tuberculosis in pre‑scapular and pre‑pectoral lymph nodes (or different lymphadenitis) in condemned carcasses. Histological processing employed routine techniques carried out at the Laboratory of Animal Pathology of the Federal Rural University of the Amazon, while the PCR assay was performed at the Bacteriology Laboratory of the Evandro Chagas Institute. Results showed that 1.96% of the histopathology samples corresponded to inflammatory processes typical of TB and that, in PCR, 4.25% of the samples had the amplification profile of the M. bovis species. These results indicate the importance of adding complementary methods to assist the sanitary inspection line and make inspection more efficient in its decisions.

RETRACTED: Neuroprotective potential of Ginkgo biloba on alteration of rat cerebellum following prenatal exposure to cyclophosphamide

The cytotoxicity of chemotherapeutic drugs is known due to its non-selective effect not only on cancer cells but also on healthy cells. This study investigated the cerebellar alteration in rats prenatally exposed to cyclophosphamide (SK, 20 mg/kg). We also evaluated the neuroprotective potential of Ginkgo biloba (GB, 80 mg/kg/day) against possible biological changes caused by SK in the cerebellar tissues. Twenty adult female rats (weighing 230–280 g, 12 weeks old) were divided into five groups: control, sham, SK, GB, and SK + GB. After mating, pregnant rats was treated with SK in the SK and SK + GB groups and GB in the GB and SK + GB groups from day 13 to day 21 of gestation. After parturition, eight female rats were randomly selected from each group. On day 32 after birth, the cerebellar tissues were dissected and then examined under light microscope using stereological and histopathological methods. Stereological findings showed that the total number of Purkinje cells and granular cells were significantly decreased in the SK group than the control group (p < 0.05). In addition, the mean volumes of molecular layer, granular layer, white matter, and cerebellum were significantly decreased in the SK group compared to the control group (p < 0.05). In the SK + GB group, the total number Purkinje cell, and granular cells, as well as the mean volumes of molecular layer, granular layer, white matter, and cerebellum were significantly increased than the SK group (p < 0.05). Histopathological evaluation also confirmed our stereological findings in the cerebellar tissues. Our results showed that prenatal exposure to SK caused significant changes in the cerebellar architectures of rats, and that GB administration significantly attenuated the deleterious effect of SK on the cerebellar tissues.

Field survey and molecular characterization of apicomplexan parasites in small mammals from military camps in Afghanistan

Small mammals are an important reservoir for causative agents of numerous infectious diseases, including zoonotic and vector-borne diseases. The occurrence of these pathogens represents a regional but permanent threat for humans and animals in general and might especially weaken military personnel and companion animals in abroad missions. In our study, small mammals collected in military camps in Afghanistan (Feyzabad, Mazar-e Sharif, and Kunduz) were investigated for the presence of apicomplexans using histopathology and molecular methods. For this purpose, well-established and newly developed real-time PCR assays were applied. A high prevalence was detected not only in house mice (Mus musculus), but also in shrews (Crocidura cf. suaveolens) and grey dwarf hamsters (Cricetulus migratorius). The molecular characterization based on the 18S rRNA gene revealed a close relationship to a cluster of Hepatozoon sp. detected in voles of the genus Microtus. Hepatozoon canis DNA was detected in one house mouse as well as in two Rhipicephalus ticks from a dog puppy. In addition, around 5% of the house mice were found to be infected with far related adeleorinids showing the highest sequence identity of 91.5% to Klossiella equi, the only published Klossiella sequence at present. For their better phylogenetic characterization, we conducted metagenomics by sequencing of two selected samples. The resulting 18S rRNA gene sequences have a length of about 2400 base pairs including an insertion of about 500 base pairs and are 100% identical to each other. Histopathology together with organ tropism and detection rates verified this sequence as of Klossiella muris. In conclusion, we documented naturally occurring protozoan stages and the additional taxonomic characterization of a well-known commensal in mice by applying a combination of different approaches. The study is of medical, social, and biological importance for ensuring human and animal health in military camps and also stresses the required awareness for the potential risk of zoonoses.

HPV genotypes in high-grade cervical lesions and invasive cervical carcinoma detected in Gabonese women

BackgroundCervical cancer is the third most common cancer among women worldwide, but particularly affects women living in sub-Saharan Africa. Screening and vaccination programs are two prevention approaches that can reduce cervical cancer incidence. However, effective vaccination campaigns require better knowledge of the prevalence of the main human papillomavirus (HPV) genotypes reported in high-grade neoplastic lesions and invasive carcinomas in women.MethodsAll samples collected in this study were processed using standard histopathological methods with haematoxylin and eosin staining of the sections. Areas with abnormal cells were then identified. The HPV genotype was determined on the DNA extracted from the same sections using nested PCR followed by amplicon sequencing and real-time PCR specific to five different HPV genotypes (16, 18, 33, 45 and 58).ResultsA total of 132 Gabonese patients with high-grade neoplastic lesions were included in this study; 81% were squamous cell carcinomas (SCC). At least one HPV was detected in 92.4% patients; HPV16 (75.4%) was the most frequent genotype, followed by HPV18, 58, 45, 33 and 35. Moreover, histological analysis showed that SCC samples had 50% and 58.2% stage III and IV tumor cells, respectively, according to the FIGO classification. Finally, 36.9% of these stage III and IV patients were less than 50 years old.ConclusionsOur results confirm the high prevalence of HPV16 and 18 genotypes among high-grade lesions in Gabonese women. This study confirms the need for a national strategy for early screening of precancerous lesions associated with a broad national vaccination program among non-sexually active women to significantly reduce the long-term cancer burden.

Protective effectiveness of anise against testicular ischemia and reperfusion injury: An experimental study in rats.

Testicular torsion is a frequently encountered clinical condition that requires urgent treatment. The aim of this study is to investigate the efficacy of Anise (Pimpinella anisum L.) in treating the pathological condition due to ischemia and reperfusion injury by using biochemical, histopathological and immunohistochemical methods. A total of 6 groups were formed with 8 male Wistar Albino rats in each group. Group 1 (n=8): control group, Group 2 (n=8): Anise aqueous solution was given orally 5 ml/kg by gavage for 30 days. Group 3 (n=8): Ischemia and Reperfusion (I/R) group, bilateral testicles were rotated 270° and reperfused after 30 minutes of ischemia. Group 4 (n=8): I/R+ Anise group, Group 5 (n=8): Anise+ I/R group and Group 6 (n=8): Anise+ I/R+ Anise group. The results of the Anise group and the Control group were similar. However, the damage in the I/R group was considerably more severe than in any of the other study groups. While it was observed that spermatogenic cells started to regenerate in the I/R+Anise group, edema and congestion were observed in the Anise+I/R group. In the Anise+I/R+Anise group, all histological findings and biochemical parameters were similar to those of the control group. It was observed that anise had protective effects in ischemia and reperfusion injury in rat testicles.

Wound healing activity of Salvia huberi ethanolic extract in streptozocin-induced diabetic rats

Objective: The aim of this study was to examine the in vivo wound healing potential of Salvia huberi Hedge (endemic to Turkey) on excision and incision wound models in diabetic rats. Method: Male Wistar albino rats, 3-4 months old and weighing 180-240g were used. The animals were randomly divided into five groups including Control, Vehicle and Fito reference, and two different concentrations (0.5% and 1% weight/weight (w/w)) of ethanol extract of Salvia huberi were investigated in both wound models on streptozocin-induced diabetic rats using macroscopic, biomechanical, biochemical, histopathological, genotoxic and gene expression methods over both seven and 14 days. Fito cream (Tripharma Drug Industry and Trade Inc., Turkey) was used as the reference drug. Results: A total of 60 rats were used in this study. Salvia huberi ointments at 0.5% and 1% (w/w) concentrations and Fito cream showed 99.3%, 99.4% and 99.1% contraction for excision wounds, and 99.9%, 97.0% and 99% contraction for incision wounds, respectively. In Salvia huberi ointments and Fito cream groups, re-epithelialisation increased dramatically by both day 7 and day 14 (p<0.05). By day 14, low hydroxyproline and malondialdehyde (MDA) levels, and high glutathione (GSH) levels were observed in the Salvia huberi ointment groups. After two application periods, damaged cell percent and genetic damage index values and micronucleus frequency of Salvia huberi ointment treatment groups were lower than Control and Vehicle groups (p<0.001). A growth factor expression reached a high level by day 7 in the Control group; in Salvia huberi-treated groups it was decreased. Conclusion: The study showed that application of Salvia huberi ointments ameliorated the healing process in diabetic rats with excisional and incisional wounds and may serve as a potent healing agent.

Wound healing activity of Salvia huberi ethanolic extract in streptozocin-induced diabetic rats

Objective: The aim of this study was to examine the in vivo wound healing potential of Salvia huberi Hedge (endemic to Turkey) on excision and incision wound models in diabetic rats. Method: Male Wistar albino rats, 3-4 months old and weighing 180-240g were used. The animals were randomly divided into five groups including Control, Vehicle and Fito reference, and two different concentrations (0.5% and 1% weight/weight (w/w)) of ethanol extract of Salvia huberi were investigated in both wound models on streptozocin-induced diabetic rats using macroscopic, biomechanical, biochemical, histopathological, genotoxic and gene expression methods over both seven and 14 days. Fito cream (Tripharma Drug Industry and Trade Inc., Turkey) was used as the reference drug. Results: A total of 60 rats were used in this study. Salvia huberi ointments at 0.5% and 1% (w/w) concentrations and Fito cream showed 99.3%, 99.4% and 99.1% contraction for excision wounds, and 99.9%, 97.0% and 99% contraction for incision wounds, respectively. In Salvia huberi ointments and Fito cream groups, re-epithelialisation increased dramatically by both day 7 and day 14 (p<0.05). By day 14, low hydroxyproline and malondialdehyde (MDA) levels, and high glutathione (GSH) levels were observed in the Salvia huberi ointment groups. After two application periods, damaged cell percent and genetic damage index values and micronucleus frequency of Salvia huberi ointment treatment groups were lower than Control and Vehicle groups (p<0.001). A growth factor expression reached a high level by day 7 in the Control group; in Salvia huberi-treated groups it was decreased. Conclusion: The study showed that application of Salvia huberi ointments ameliorated the healing process in diabetic rats with excisional and incisional wounds and may serve as a potent healing agent.