Cytological Studies Research Articles

Retrospective Study of Fine Needle Aspiration Cytology of Patients with Thyroid Nodule in Benghazi in the Period of (2021-2023)

Background: Thyroid nodules constitute a common endocrine disorder, these nodules could be solitary, multiple, cystic, or solid. Increasing age, female gender, iodine deficiency, and history of thyroid radiation all are high risk for thyroid nodules. FNAC representing the most cost-effective diagnostic tool used in the assessment of nodules of the thyroid. Objective of the study: The aim of this study is to evaluate the result of fine needle aspiration cytology (FNAC) and assess thyroid status using of thyroid hormone obtained from patients with thyroid nodules. Materials and methods: this is a retrospective study, includes collection of data from available cases of thyroid nodules underwent fine needle aspiration between 2021 to 2023, data regarding age, sex, TFT, USS, FNAC was collected from the archive of endocrine out patients clinic of Benghazi Medical Center and from endocrine clinic of Benghazi diabetic center. The analysis was done using the SPSS program. Result: A total of 60 cases were studied, Data was collected from 60 patient’s files for years between 2021 and 2023. The mean age of the patients with FNAC selected for this study was 49.65, and ranged from 21-80 years. The gender distribution shows female predominance in which female cases were 53 (98.33%) while only one case (1.67%) was reported among men. The size of nodules in our cases range from 1cm to 4.5 cm. The mean size of nodules is 1.89cm. In this study the largest group of nodules located in right lobe (21) cases then left lobe (20) cases then both (17) cases and in isthmus only 2 cases. Most of the cases thyroid function test result shows an euthyroid status (38) cases. FNAC of thyroid nodule showed that most of patient’s result diagnosed as benign nodules (47) cases. Conclusion: FNAC is a key tool in diagnosis of thyroid nodule as it has safe, minimally invasive and cost effective diagnostic role.

Current epidemiological characteristics of cervical cancer in the Russian Federation

Background. The prevalence of cervical cancer (Cca) in Russian women makes it crucial to study modern epidemiological characteristics and adjust control measures accordingly.The aim. To analyze trends in the incidence of cervical cancer in the Russian Federation.Materials and methods. A retrospective epidemiological study was conducted on cervical cancer incidence in Russian Federation from 2008 to 2021. The materials of annual statistical data “Malignant neoplasms in Russia (morbidity and mortality)” and “Status of oncological assistance to the population of Russia” (2008–2022), data of the federal statistical reporting form No. 5 “Information about preventive vaccinations” (2021), data on the number of studies conducted (cytological study and HPV-test based on RT-PCR) in the clinical diagnostic laboratory of Central Research Institute of Epidemiology of Rospotrebnadzor (2013–2021) were used.Results and discussion. The trends of Cca incidence varied depending on the women’s age. Starting in 2016, there was a stabilization of the level of morbidity in women aged 30–49 with the subsequent decrease from 2019 due to the strengthening of secondary prevention measures, including the active introduction of HPV-test into the diagnostic schemes conducted in the Russian Federation. Increased control of cervical cancer caused a rise in the proportion of cases diagnosed at an early stage (carcinomas in situ and I–II stages Cca), the increase of detection rate of in situ carcinomas per 100 cases of Cca, the growth of the proportion of carcinomas in situ to I–II stages Cca, rise of the percentage of the cases detected actively, decrease of the number of Cca cases detected at later stages and one-year mortality index.Conclusion. The epidemiological analysis of cervical cancer incidence shows that preventive measures are effective and that there is a need to intensify and increase their coverage.

SURG-22. BRAIN METASTASIS FROM ADVANCED GASTRIC CANCERS: HIGH INCIDENCE OF LEPTOMENINGEAL DISSEMINATION AND ITS CLINICAL IMPACT ON SURVIVAL

Abstract OBJECTIVE Advanced gastric cancer is one of the most common and fatal cancers worldwide, and due to its highest incidence rate in Eastern Asian, it is steadily considered important. However, brain metastasis from advanced gastric cancer is very rare, and its incidence is reported as less than 1% of patients with primary advanced gastric cancer. There have been very few studies describing on the incidence and clinical outcome of patients with brain metastasis from advanced gastric cancers. METHOD We retrospectively analyzed 128 patients diagnosed as brain metastasis from advanced gastric cancers, and then underwent treatment from 2005 to 2021 April in our institution. Patients’ survival and its associated prognostic factors were evaluated. Especially, subgroup analysis on patients with leptomeningeal dissemination (LMD) was performed. RESULT Of total 128 patients of advanced gastric cancer with brain metastasis, 74 patients (58%) were diagnosed with LMD. Overall survival of LMD group was significantly shorter than that of non-LMD group (p<0.001). Patients treated with tumor removal (TR) followed by whole brain radiotherapy (WBRT) showed superior overall survival than patients with WBRT alone (p=0.002). Among 74 LMD patients, those who underwent WBRT combined with intrathecal methotrexate (IT MTX) showed better outcome than patients with WBRT alone (p=0.025). Specifically, for the 51 pure LMD patients without parenchymal brain metastasis, the superiority of adjuvant IT TMX on survival outcome was more apparent (p=0.011). CONCLUSION The incidence of LMD has been underestimated as 58% of patient with brain metastasis from advanced gastric cancers presented LMD on radiological and cytological studies. If the brain tumor is surgically resectable, TR followed by WBRT can be the first choice of treatment provides survival benefit. Patients with LMD are most benefited from IT MTX combined with WBRT.

Potential treatment approaches for malignant peritoneal mesothelioma: in vivo and in vitro experimental study of natural killer cell immunotherapy.

Malignant peritoneal mesothelioma (MPM) is a rare primary malignant tumor with an extremely poor prognosis that currently lacks effective treatment options. This study investigated the in vitro and in vivo efficacy of natural killer (NK) cells for treatment of MPM. An in vitro study was conducted to assess the cytotoxicity of NK cells from umbilical cord blood to MPM cells with the use of a high-content imaging analysis system, the Cell Counting Kit-8 assay, and Wright-Giemsa staining. The level of NK cell effector molecule expression was detected by flow cytometry and enzyme-linked immunosorbent assays. The ability of NK cells to kill MPM cells was determined based on live cell imaging, transmission electron microscopy, and scanning electron microscopy. An in vivo study was conducted to assess the efficacy and safety of NK cell therapy based on the experimental peritoneal cancer index, small animal magnetic resonance imaging, and conventional histopathologic, cytologic, and hematologic studies. NK cells effectively killed MPM cells through the release of effector molecules (granzyme B, perforin, interferon-γ, and tumor necrosis factor-α) in a dose- and density-dependent manner. The NK cell killing process potentially involved four dynamic steps: chemotaxis; hitting; adhesion; and penetration. NK cells significantly reduced the tumor burden, diminished ascites production, and extended survival with no significant hematologic toxicity or organ damage in NOG mice. NK cell immunotherapy inhibited proliferation of MPM cells in vitro and in vivo with a good safety profile.

CytoNet: an efficient dual attention based automatic prediction of cancer sub-types in cytology studies

Computer-assisted diagnosis (CAD) plays a key role in cancer diagnosis or screening. Whereas, current CAD performs poorly on whole slide image (WSI) analysis, and thus fails to generalize well. This research aims to develop an automatic classification system to distinguish between different types of carcinomas. Obtaining rich deep features in multi-class classification while achieving high accuracy is still a challenging problem. The detection and classification of cancerous cells in WSI are quite challenging due to the misclassification of normal lumps and cancerous cells. This is due to cluttering, occlusion, and irregular cell distribution. Researchers in the past mostly obtained the hand-crafted features while neglecting the above-mentioned challenges which led to a reduction of the classification accuracy. To mitigate this problem we proposed an efficient dual attention-based network (CytoNet). The proposed network is composed of two main modules (i) Efficient-Net and (ii) Dual Attention Module (DAM). Efficient-Net is capable of obtaining higher accuracy and enhancing efficiency as compared to existing Convolutional Neural Networks (CNNs). It is also useful to obtain the most generic features as it has been trained on ImageNet. Whereas DAM is very robust in obtaining attention and targeted features while negating the background. In this way, the combination of an efficient and attention module is useful to obtain the robust, and intrinsic features to obtain comparable performance. Further, we evaluated the proposed network on two well-known datasets (i) Our generated thyroid dataset (ii) Mendeley Cervical dataset (Hussain in Data Brief, 2019) with enhanced performance compared to their counterparts. CytoNet demonstrated a 99% accuracy rate on the thyroid dataset in comparison to its counterpart. The precision, recall, and F1-score values achieved on the Mendeley Cervical dataset are 0.992, 0.985, and 0.977, respectively. The code implementation is available on GitHub. https://github.com/naveedilyas/CytoNet-An-Efficient-Dual-Attention-based-Automatic-Prediction-of-Cancer-Sub-types-in-Cytol.

Repetitive transcranial magnetic stimulation alleviates motor impairment in Parkinson’s disease: association with peripheral inflammatory regulatory T-cells and SYT6

BackgroundRepetitive transcranial magnetic stimulation (rTMS) has been used to treat various neurological disorders. However, the molecular mechanism underlying the therapeutic effect of rTMS on Parkinson’s disease (PD) has not been fully elucidated. Neuroinflammation like regulatory T-cells (Tregs) appears to be a key modulator of disease progression in PD. If rTMS affects the peripheral Tregs in PD remains unknown.MethodsHere, we conducted a prospective clinical study (Chinese ClinicalTrials. gov: ChiCTR 2100051140) involving 54 PD patients who received 10-day rTMS (10 Hz) stimulation on the primary motor cortex (M1) region or sham treatment. Clinical and function assessment as well as flow cytology study were undertaken in 54 PD patients who were consecutively recruited from the department of neurology at Zhujiang hospital between September 2021 and January 2022. Subsequently, we implemented flow cytometry analysis to examine the Tregs population in spleen of MPTP-induced PD mice that received rTMS or sham treatment, along with quantitative proteomic approach reveal novel molecular targets for Parkinson's disease, and finally, the RNA interference method verifies the role of these new molecular targets in the treatment of PD.ResultsWe demonstrated that a 10-day rTMS treatment on the M1 motor cortex significantly improved motor dysfunction in PD patients. The beneficial effects persisted for up to 40 days, and were associated with an increase in peripheral Tregs. There was a positive correlation between Tregs and motor improvements in PD cases. Similarly, a 10-day rTMS treatment on the brains of MPTP-induced PD mice significantly ameliorated motor symptoms. rTMS reversed the downregulation of circulating Tregs and tyrosine hydroxylase neurons in these mice. It also increased anti-inflammatory mediators, deactivated microglia, and decreased inflammatory cytokines. These effects were blocked by administration of a Treg inhibitor anti-CD25 antibody in MPTP-induced PD mice. Quantitative proteomic analysis identified TLR4, TH, Slc6a3 and especially Syt6 as the hub node proteins related to Tregs and rTMS therapy. Lastly, we validated the role of Treg and rTMS-related protein syt6 in MPTP mice using the virus interference method.ConclusionsOur clinical and experimental studies suggest that rTMS improves motor function by modulating the function of Tregs and suppressing toxic neuroinflammation. Hub node proteins (especially Syt6) may be potential therapeutic targets.Trial registrationChinese ClinicalTrials, ChiCTR2100051140. Registered 15 December 2021, https://www.chictr.org.cn/bin/project/edit?pid=133691Graphical rTMS is a safe and non-invasive method for Parkinson's disease. In this study, we showed the proportion of CD4+CD25+CD127- regulatory T-cells (Tregs) in the peripheral blood was significantly increased after rTMS treatment. Similar effects of rTMS treatment were verified in MPTP-induced PD mice. Proteomic analysis and RNA interference analyses identified TLR4, TH, Slc6a3 and especially Syt6 as hub node proteins that can be modulated by rTMS therapy in PD.

Dynamics of the effect of local treatment on the frequency of nuclei with perinuclear vacuole in the cytogram of buccal epithelium in patients with the erosive and ulcerative form of the lichen planus in the oral cavity mucosa lining

INTRODUCTION. The use of cytologic studies allows evaluating the dynamics of the local treatment conducted of the EUF (L43.82) LP OCML (Erosive and Ulcerative Form of the Lichen Planus in the Oral Cavity Mucosa Lining) in a comprehensive way.AIM. Evaluation of the effect of local the effect of local treatment on the frequency of nuclei with perinuclear vacuole in the cytogram of buccal epithelium of the reticular mesh surface, hyperemic zone and erosive and ulcerative elements in patients with the erosive and ulcerative form of the flat lichen of the oral mucosa (L43.82).MATERIAL AND METHODS. 86 patients with the EUF (L43.82) LP OCML participated in the study. The number of nuclei with perinuclear vacuole was assessed in cytologic preparations obtained from buccal epithelium, which was a reliable criterion to confirm the nature of chronic inflammation. The dynamics of cells with necrosis in the cytogram of buccal epithelium was evaluated using nonparametric criteria as follows: Mann-Whitney test (comparison between subgroups and with the experimental group), Wilcoxon test (to compare within subgroups before and after treatment). Significant differences did not exceed 0.05.RESULTS. In the course of the treatment conducted in the cytogram of buccal epithelium there was a decrease in the frequency of nuclei with perinuclear vacuole of the reticular mesh surface and erosive and ulcerative elements in patients of the main clinical groups 1 and 2(p< 0.001). When analyzing the frequency of nuclei with perinuclear vacuole in the cytogram of buccal epithelium taken from the surface of erosions and ulcers, there were significant differences before and after treatment according to the Wilcoxon test. In the clinical group using the treatment method according to federal guidelines, the frequency of nuclei with perinuclear vacuole was observed at a significance level of p< 0.05, and when using the local treatment complex proposed respectively at p< 0.001.CONCLUSION. Thus, in the cytogram of buccal epithelium according to the Wilcoxon test before and after the local treatment proposed, there was a more evident decrease in the number of nuclei with perinuclear vacuole than with the use of federal guidelines.

Macrophage hyperfunction as a cellular trigger for fibroplastic changes in internal organs in pneumoconiosis among miners

Introduction. The significant prevalence of anthracosilicosis and silicosis, characterized by the rapid development of pneumofibrosis in coal mine workers, is a serious socially significant problem worldwide. A key component of the pathogenesis of pneumoconiosis is the macrophage reaction to fibrogenic dust, which is a complex immunological response, with prolonged and intense exposure leading to chronic inflammation in various organs and systems, fibrosis and a progressive decrease in their functions. A deep understanding and evaluation of the macrophage reaction as a potential biomarker for the diagnosis of lung damage in miners will allow us to develop more effective strategies to combat dust damage to organs and systems. The study aims to consider macrophage hyperfunction as a trigger mechanism for fibroplastic changes in internal organs in pneumoconiosis in miners. Materials and methods. Using light microscopy, specialists conducted extended cytological, histological and immunohistochemical studies of bronchial smears and autopsy material (fragments of bronchi, lungs, heart, liver and kidneys) of 50 miners of Kuzbass coal enterprises who died simultaneously as a result of a man-made disaster. Morphometric analysis of histological structures with measurement of linear dimensions was used for a comprehensive assessment of pathomorphological changes. Results. There was a sharp increase in the number and size of actively phagocytic alveolar macrophages loaded with dust particles in the lungs of miners. Pathological changes in internal organs were characterized by pronounced fibrosis in the peribronchial and perivascular zones, stroma with deposition of dust particles and a pronounced macrophage reaction. In a group of miners with more than 15 years of experience in the central and distal parts of the lungs, in the intermuscular zones of the myocardium, the portal tracts of the liver, and the central segments of the kidneys, there was a constant increase in the expression of immunohistochemical markers CD14, CD34, actin and vimentin, which can be used as diagnostic markers for the identification of macrophage inflammation and fibrosis, will allow to assess the degree of severity of the pathological process. A characteristic feature of the miners' long experience of dust exposure was the presence of progressive pronounced perivascular sclerosis extending beyond the vascular histione. Experts found fibroblastic cells with positive expression of CD14, CD31 and CD34 in the sclerotic zones. Limitation. The absence of cytogenetic and electron microscopic examination, which is expected in the future in conjunction with the expansion of the range of immunohistochemical research methods. Conclusion. The results of extended pathomorphological and cytological studies emphasize the importance of an integrated approach to the diagnosis of occupational diseases in the coal industry. The use of a combination of these methods will allow timely detection of macrophage inflammation and fibrous process in dust pathology, specific changes at the cellular and tissue levels, as well as to assess the severity of the pathological process. Ethics. The study of autopsy material was based on a secondary examination of blocks and ready-made histological micro-preparations (glasses) of the material of the Bureau of Forensic Medical examination of Novokuznetsk, Osinniki, Prokopyevsk. The studies of pathomorphological material were carried out strictly on the basis of Federal Law No. 323-FZ dated 11/21/2011 "On the Basics of Protecting the Health of Citizens in the Russian Federation", in particular Article 67 "Conducting pathological anatomical autopsies", Federal Law No. 8-FZ dated 12.01.1996 "On Burial and Funeral Business" (Article 5, pp. 1, 2), as well as in accordance with the Order of the Ministry of Health of the Russian Federation dated 06.06.2013 No. 354n "On the procedure for conducting pathological anatomical autopsies", Order of the Ministry of Health of the Russian Federation dated 03/24/2016 No. 179n "On the Rules for conducting pathological anatomical studies".

Abnormal expression of LiASY1 and LiDMC1 is the main cause of sterility in F1 hybrids developed from Lagerstroemia speciosa and Lagerstroemia indica ‘Ebony Embers’

Lagerstroemia × ‘Xiaoxiang Huaxiu’ (an F1 hybrid of Lagerstroemia speciosa and Lagerstroemia indica ‘Ebony Embers’) exhibits significant sterility. Cytological studies have revealed numerous abnormal chromosomal behaviors during the meiotic division of pollen mother cells in L. × ‘Xiaoxiang Huaxiu’. Transcriptome sequencing identified that LiDMC1 and LiASY1 were expressed at very low levels in ‘Xiaoxiang Huaxiu’. To further verify whether the abnormal expression of LiDMC1 and LiASY1 was the fundamental cause of sterility in ‘Xiaoxiang Huaxiu’, we cloned the full-length gene sequences of LiDMC1 and LiASY1 from ‘Ebony Embers’. Using VIGS (virus-induced gene silencing) technology, pTRV1 and pTRV2 gene silencing vectors were employed to silence LiDMC1 and LiASY1 in the paternal parent ‘Ebony Embers’. The results indicated that in the double-silenced lines of ‘Ebony Embers’, the cytoplasm sizes of different tetrads during the tetrad stage varied significantly. There was a notable proportion of polyads, binucleate cells with micronuclei, and pentads. Additionally, pollen diameter varied significantly, closely resembling the sterility observed in ‘Xiaoxiang Huaxiu’. Pollen viability in the double-silenced ‘Ebony Embers’ decreased by 22.8%. This study demonstrates that the low expression of LiDMC1 and LiASY1 leads to abnormal microspore development in ‘Ebony Embers’, and their low expression is the cause of sterility in ‘Xiaoxiang Huaxiu’.

Study the Occurrence of Mycobacterium Bovis in Tuberculosis of Peripheral Lymph Nodes and its Effect on the Course of the Disease.

The incidence of tuberculosis caused by Mycobacterium bovis, not only the pulmonary form, but also the form developing in the extrapulmonary organs, is also increasing from year to year. Despite the large number of EPTB, TBPLN occupies a leading place among diseases of this type and the study of its pathogenic strains is an urgent task in ensuring the effectiveness of treatment. In this regard, the main purpose of the presented manuscript is to determine the frequency of M. bovis in TBPLN, its effect on the development and course of the disease, as well as the effectiveness of treatment. For this purpose, for the first time, the features of education that occur in patients in peripheral lymph nodes using instrumental methods of ultrasound, computed tomography, and magnetic resonance imaging have been identified. In subsequent studies, 110 patients with peripheral lymph node pathology were diagnosed with TBPLN by detecting mycobacteria in pathological material using general hematological, microbiological and gene-molecular (Gene Xpert) methods. In order to ensure the high effectiveness of drugs used for medicinal purposes, strains of the pathogen were detected using histological, cytological studies, BCG test and specific analyzes such as Diaskintest, Quantiferon test, immunological tests. The study showed that about 80% of patients had M bovis in the overall assessment, 76.4% of patients were sensitive to rifampicin, 9.1% of patients had rifampicin-resistant bacteria, and 14.5% of patients did not have mycobacteria. Therapeutic measures were carried out in 2 different modes, such as standard and individual or with replacement, when all patients were divided into 2 groups. During the period from the 56-day intensive phase of standard treatment to the 84-day intensive phase, a total of 40 patients had a sharp decrease in lymph nodes, elimination of purulent inflammation, and after a while 22 patients in this group had a relapse. In the individual treatment regimen, Levofloxacin and linezolid were used instead of pyrazinamide. While the effectiveness of treatment was achieved in 48 patients of group II after 56 and 84 days of the intensive phase, relapses after a certain time were observed in only 6 patients. When choosing an individual treatment regimen in patients diagnosed with M. bovis, a decrease in relapses to 11.5% is achieved. When M. bovis is detected, an individual scheme of antibacterial treatment of tuberculosis is selected, in which, instead of pyrazinamide, it is recommended to choose one of the reserve lines, depending on the sensitivity of the pathogen to drugs.

Using express cytological method for diagnosis of sentinel lymph nodes in breast cancer patients

Objective: to assess the informativity of the use of the rapid cytological method in the study of sentinel lymph nodes in patients with breast cancer. Material and methods. The study involved 46 patients from 30 to 85 years old with verified breast cancer. On the eve of the operation, about 150-200 MBq of the 99 mTc-Nanotope radiopharma-ceutical was peritumorally injected into the parenchyma of the affected mammary gland. After 1.5-2 hours, lymphos-cintigraphy was performed, followed by SPECT/CT, in which sentinel lymph nodes are zualized. The next day after the administration of the radiopharmaceutical, during surgery, sentinel lymph nodes were found using a g-detector. Further, the surgeon removed the node (or nodes) and sent them for express cytological examination. To check the informativity of this method, a comparison was made with the "gold standard" of morphological diagnostics — with histological examination of postoperative material. Results. An express cytological study revealed the presence of metastases in the sentinel lymph nodes in 12 patients, which significantly influenced the further tactics. It was found that the cytological method of the study is not inferior to the histological one, the sensitivity and specificity of the method is 100%. When comparing the results of rapid cytological and planned histological studies, there were no discrepancies in diagnoses by the state of the lymph nodes. Conclusion. Rapid cytological examination of sentinel limphatic nodes informative method for sentinel lymph node detection in breast cancer patients.

Study of the role of polymorphic variants G919A and A2039G of the follicle stimulating hormone receptor gene FSHR in the genesis of male infertility

Aim. To determine the distribution of genotypes of polymorphic variants G919A and A2039G of the gene FSHR (follicle-stimulating hormone receptor) among men with azoospermia. Methods. DNA from peripheral blood leukocytes was isolated and purification using a modified salting out method. Extracted DNA was amplified by Polymerase chain reaction (PCR). The PCR products were subsequently digested with the restriction enzyme for identify polymorphic variants of the follicle-stimulating hormone receptor gene FSHR. Electrophoresis of PCR products was performed in a 2 % agarose gel. Results. Given that idiopathic infertility is overwhelmingly caused by genetic factors, it seemed necessary to conduct a set of cytological and molecular genetic studies in a group of men with azoospermia. The genetic component was verified in 28 men with azoospermia, which is 40 % of all subjects. Molecular genetic studies were performed and the distribution of genotypes of polymorphic variants A919G and A2039G of the FSHR gene among men with azoospermia was determined. Conclusions. A slightly higher frequency of homozygous genotypes GG of polymorphic variants A919G and A2039G of the follicle stimulating hormone receptor gene FSHR was found among men with azoospermia compared to the group of fertile men.

In Planta Androgenesis (Paternal Origin) in Saacharum X Erianthus Intergeneric Hybrid

The present investigation was carried in crosses made between Saccharum species hybrid clones i.e. Co 419, Co 7201 and Co 7224 (2n = 108-112) and Erianthus arundinaceus viz. IK 76-91 (2n = 60) and IK 76-99 (2n = 60) for introducing better ratoon ability, vigour, wide ecological adaptability, extensive root system, drought and water logging resistance, high biomass and disease resistance into modern sugarcane cultivars during 2020-23 at VSI-Sugarcane Breeding Centre, Amboli. Parental apomixis (paternal origin) has been recently noticed and it is unique in Saccharum X Erianthus intergeneric cross seedlings. A total of fifty-one seedlings were obtained from above three intergeneric crosses and out of these eighteen seedlings were reported as androgenetic seedlings as the female act as a surrogate mother. At the early stages, seed germination of seedlings was found two types i.e., one type resembling Saccharum and other resembling Erianthus with deep purple coloration. Morphologically androgenetic seedlings were similar to the male parents IK 76-91 and IK 76-99 with tall large tufted grass like clumps, vegetative stem with thick reproductive buds, root zone narrow and only one row of root eyes, the lamina gradually passes into leaf sheath with no auricle and dewlaps. Cytological studies were made in these seedlings and during metaphase the somatic chromosome number was observed to be 2n = 60 rather than expected 2n = 84-90. This paternal origin may be due to the ability of Erianthus male gametes to produce an embryo within the seed tissues of Saccharum species hybrids but without genetic contribution of the seed. Thus, in planta androgenesis is achieved through the combination of embryogenic behavior of male gametes which has entered in the ovule and the ability of ovules to act as a surrogate mother. They could derive either from the fusion of two male gametes or from the early natural diplodization of the haploid embryo. Such homozygous true breeding seedlings of Saccharum X Erianthus with hybrid vigour are extremely important in crop improvement. This system provides an unparalleled opportunity to shorten the breeding cycle and fix agronomic traits in the homozygous state. These androgenetic plants with Saccharum cytoplasm and Erianthus genome are male sterile making them highly useful for further hybridization and sugarcane improvement programme.

Feasibility of Using Needle Rinse Fluid for Cobas Human Papillomavirus (HPV) Assay in Diagnosing HPV+ Oropharyngeal Cancer with Neck Lymph Node Aspiration.

Human papillomavirus (HPV) is a crucial prognostic factor in oropharyngeal cancer (OPC). p16 is a surrogate marker for diagnosing HPV+ OPC, however it is not direct evidence of HPV existence. The purpose of our study was to evaluate an HPV DNA test-Cobas HPV assay-in diagnosing HPV+ OPC through neck lymph node aspiration. Patients with suspected neck mass who received fine needle aspiration (FNA) or core needle biopsy (CNB) at the National Taiwan University Hospital between January 2018 and December 2022 were reviewed. Besides routine cytology and pathology study, needle rinse fluid was collected for the Cobas HPV assay to detect high-risk HPV. We analyzed 137 patients with suspected lymph nodes, 32 (23.4%) of whom were HPV+ OPC patients and 105 (76.6%) of whom had non-HPV-related disease. FNA was performed in 31 patients and CNB was performed in 106 patients, according to the size and necrosis status of the lymph nodes. For diagnosing HPV+ OPC, CNB combined with p16 immunohistochemistry staining showed sensitivity of 93.3%, specificity of 97.8%, positive predictive value (PPV) of 87.5%, negative predictive value (NPV) of 98.9%, and accuracy of 97.2%. On the other hand, for the needle rinse Roche Cobas HPV assay, the test showed sensitivity of 96.9%, specificity of 100%, PPV of 100%, NPV of 99.1%, and accuracy of 99.3%. Compared with p16 IHC staining, the Cobas HPV test showed better PPV with statistical significance (p=0.04). The Cobas HPV assay is a US FDA-approved, highly automated, and readily used technique to directly detect the presence of high-risk HPV. We recommend utilizing the Cobas HPV assay in combination with routine cytology or histopathology examination in the work-up of neck lymphadenopathy.

Microplastic exposure reduced the defecation rate, altered digestive enzyme activities, and caused histological and ultracellular changes in the midgut tissues of the ground beetle (Blaps polychresta)

Concerns about microplastic (MP) pollution in terrestrial systems are increasing. It is believed that the overall amount of MPs in the terrestrial system could be 4–23 times higher than that in the ocean. Agricultural ecosystems are among the most polluted areas with MPs. Terrestrial organisms such as ground beetles, will be more vulnerable to MPs in various agricultural soil types because they are common in garden and agricultural areas. Therefore, this work aims to assess for the first time the potential adverse effects of chronic exposure for 30 days of ground beetles to a field-realistic concentration of 2 % (w/w) of three different irregularly shaped MPs polymers: Polystyrene (PS), polyethylene terephthalate (PET), and polyamide 6 (PA; i.e., nylon 6) on their health. The results showed no effect on beetle survival; nevertheless, there was a decrease in beetle defecation rate, particularly in beetles exposed to PS-MPs, and a change in the activity of midgut digestive enzymes. The effects on digestive enzymes (amylase, protease, lipase, and α-glucosidase) were polymer and enzyme specific. Furthermore, histological and cytological studies demonstrated the decomposition of the midgut peritrophic membrane, as well as abnormally shaped nuclei, vacuolation, disordered microvilli, necrosis of goblet and columnar cells, and necrosis of mitochondria in midgut cells. Given the importance of ground beetles as predators in most agricultural and garden settings, the reported adverse impacts of MPs on their health may impact their existence and ecological functions.

A jasmonate-mediated regulatory network modulates diurnal floret opening time in rice.

Diurnal floret opening time (DFOT) is a pivotal trait for successful fertilization and hybrid breeding in rice. However, the molecular mechanism underlying this trait is poorly understood in rice. In this study, we combined the cytological, genetic and molecular studies to demonstrate that jasmonic acid (JA) regulates DFOT in rice through modulating the turgor and osmotic pressure of the lodicules. We show that lodicules undergo dramatic morphologic changes, accompanied by changes in water and sugar contents during the process of floret opening. Consistently, a large set of genes associated with cell osmolality and cell wall remodeling exhibits distinct expression profiles at different time points in our time-course transcriptomes of lodicules. Notably, a group of JA biosynthesis and signaling genes is continuously upregulated, accompanied by a gradual increase in JA accumulation as floret opening approaching. Furthermore, we demonstrate that the JA biosynthesis gene OsAOS1 is required for endogenous JA biosynthesis in lodicules and promoting rice DFOT. Moreover, OsMYC2, a master regulator of JA signaling, regulates rice DFOT by directly activating OsAOS1, OsSWEET4, OsPIP2;2 and OsXTH9. Collectively, our findings establish a core regulatory network mediated by JA for modulating rice DFOT and provide effective gene targets for the genetic improvement of DFOT in rice.

Application of the Milan System for Reporting Salivary Gland Cytopathology with Histopathological Correlation

Background: Fine-needle aspiration (FNA) is widely accepted as an efficient first line diagnostic of salivary gland lesions.Intratumoral heterogeneity and morphological overlap existing between salivary gland tumors makes it rigorous andchallenging in the diagnosis by FNA. The Milan System for Reporting Salivary Gland Cytopathology (TMSRSGC) is a tieredrisk-stratification scheme designed to standardize reporting and facilitate decision making. The objective of this study is toevaluate the efficacy of The Milan system for reporting salivary gland cytopathology with histopathological correlation.Method: A prospective study of salivary gland cytology was performed over a 1-year period at Department of PathologyBPKIHS. Cases were categorized as per the MSRSGC and histopathological correlation was obtained. The risk ofmalignancy were calculated for all diagnostic categories.Result: A total of 30 salivary gland cytology with histopathological follow up were evaluated. FNA had a sensitivity,specificity, diagnostic accuracy, PPV and NPV of 72.7%, 100%, 88.88%, 100% and 84.2% respectively. Good agreementwas seen between cytological and histopathological diagnosis.Conclusion: The Milan System for Reporting Salivary Gland Cytopathology provides an effective standardizeddiagnostic framework which aids in better clinicopathological communication and improves the quality, clarity as well asreproducibility of cytological diagnosis of salivary gland lesions.

Breaking Biomphalaria black box by in situ revelation of fluorescent Schistosoma mansoni parasites

Tissue clearing is an old-fashioned method developed in the 1900's and used to turn an opaque biological object into a 3D visualizable transparent structure. Developed and diversified over the last decade, this method is most of the time applied to mammals' tissues, and especially mouse and human tissues for cytological, histological and pathophysiological studies. Through autofluorescence, immunofluorescence, in situ hybridization, intercalating agents, fluorescent transfection markers or fluorescent particle uptake, optically cleared samples can be monitored to discover new biological structures and cellular interactions through 3D-visualization, which can be more challenging in some extend through classical histological methods. Most of the tissue clearing procedures have been developed for specific applications like endogenous fluorescence visualization, immunolabeling or for revealing specific organs. Thus, choosing the adapted protocol may be empirical for non-model species, especially for mollusks for which very little related literature is available. Herein, we suggest an effective optical tissue clearing procedure for the freshwater snail Biomphalaria glabrata, known as the intermediate host of the human parasite Schistosoma mansoni. This clearing procedure involves solvents with a minimal toxicity, preserves the endogenous fluorescence of labeled parasites inside snail tissues and is compatible with an immunolabeling procedure.

Cellular changes and PR-1 gene expression accompanying infection-related morphogenesis and cell progression during invasive growth Pyricularia oryzae pathotype Triticum in resistant and susceptible wheat cultivars

Pyricularia oryzae pathotype Triticum (PoT) causes Wheat Blast, a devastating crop disease present in many wheat-producing countries. With the aim to contribute with the understanding of the mechanisms underlying pathogenesis of the fungus on wheat (Triticum aestivum), cytological and molecular studies were performed here.Susceptible and resistant interactions of wheat-PoT in cultivars of different levels of resistance were investigated by cellular changes and the molecular responses of wheat leaves according PR-1 expression. There was significant difference between susceptible (Apogee-PY15-PY34, Buck, MSINTA-PY15) and resistant (Buck-PY34, MSINTA-PY34) interactions in the responses tested. The two isolates of PoT analysed showed different ability to infect wheat plants at the cellular level during early tissue penetration analysed and documented by epi-fluorescence microscopy, patterns of PR-1 genes, and wheat plant defense reaction.Microscopic time-course analysis (24, 48, 72 and 96 h after inoculation) of individual interaction sites per leaf revealed different leaf invasion strategies of the more and less aggressive PoT isolates during the first stages of the course of pathogenesis.The results and conclusions of the present study provides the first evidence that different PR-1 spatial and temporal expression patterns occur in the early infective process of wheat-PoT, underlying two different defence mechanisms in plants according susceptible or resistant interactions. Thus, the results here laid a theoretical foundation for the future control of wheat blast using the different pattern of PR-1 gene as markers for disease resistance during the first stages of infection providing a significant contribution to a more efficient selection of wheat genotypes in breeding studies.

THE CHEMISTRY OF THE GENUS STEVIA (ASTERACEAE)

The morphological characteristic and the cytological studies made on the genus Stevia are described. Take into aécount that the chemistry of the genus might be useful for a more precise delimitation of their species; to establish relationships between chemistry, morphology and chromosomic number and to establish relationships with other genera of the tribe, the review of the isolated compounds from 54 species and 8 varieties already studied up 1997 is made.