Areas Of Necrosis Research Articles

Pathology and Molecular Diagnosis of Respiratory Disease Outbreak due to PCV-2 in a Pig Farm in Goa, India

Background: Pig farming is an important source of income and nutrition for rural farmers in Goa. Pig production is threatened by many infectious diseases. The present study reports a respiratory disease outbreak in a total of 200 pigs including adults and piglets with 40% mortality in a pig herd of Large white Yorkshire and crossbreds in South Goa during the winter period in January 2022. Method: The farm was visited and recorded clinical signs and post mortem findings and and samples were collected for bacterial isolation, PCR confirmation of bacterial and viral pathogens and histopathology. Result: The affected pigs showed high fever, reduced feed intake, staggering gate, huddling, difficulty in breathing, cough, and brown to greenish diarrhea followed by death within a span of 3 weeks. Major gross lesions were non-collapsed lungs with severe congestion and localized areas of consolidation, severe congestion of viscera, and enlargement, and presence of multifocal areas of necrosis in the liver. Histopathology of lungs focal or diffuse bronchointerstitial pneumonia and the lymphoid organs showed lymphoid depletion. Pasteruella multocida could be isolated from heart blood and tissues from 3 cases and PCR of the tissue DNA confirmed the presence of P. multocida and Porcine Circo Virus-2 (PCV-2) infection. The study confirms the presence of PCV-2 infection in pig herds in Goa for the first time. The acute mortality can be attributed to the co-infection of the herd with PCV-2 and P. multocida and sudden change in weather patterns with prevailing severe cold conditions during time of outbreak. Sequencing and Phylogenetic analysis of the PCV-2 ORF-2gene showed the PCV-2 virus from Goa is more related to isolates from southern Indian states.

The influence of perivascular tissue on lateral thermal expansion during bipolar vessel sealing

BackgroundLateral heat propagation has been an unavoidable effect of bipolar sealing with the risk of damage to surrounding structures. It is presently unknown whether leaving the perivascular tissue in situ may be advantageous in the sense of an isolation effect. Material and methodsTwo groups were formed from ex vivo carotid specimens. Group A (n = 10) consisted of carotid artery with the perivascular connective tissue in place (mean preparation diameter: 10.57 ± 0.16 mm) and group B (n = 10) of skeletonized carotids (mean vessel diameter: 5.21 ± 0.12 mm). All specimens were fixed on a plastic plate and mounted vertically in a holder. Sealing was performed perpendicular to the axis of the specimens. The temperature during the sealing process was recorded by a thermal camera. Group comparison was performed by a nonparametric test and significance was set at p < 0.05. ResultsMean sealing time in group A was 3.71 ± 0.37 s compared to 3.42 ± 0.37 s (p = 0.009) in group B. The maximum temperature in the middle of the jaws was significantly different. Group A had a temperature of 71.4 ± 3.9 °C and group B had a temperature of 91.4 ± 7.4 °C (p < 0.0001). RILATE risk scores (percent of necrotic zone in relation to potential area of necrosis) at both upper and lower sides of instrumental jaws were significantly different. For group A, it was 14.9 ± 1.6 at the upper side of jaws, 20.4 ± 2.63 at the lower side of jaws and for group B, it was 21.9 ± 3.5 at the upper side of jaws, 30.2 ± 6.2 at the lower side of jaws. ConclusionPerivascular connective tissue acts as an insulator with respect to lateral heat propagation. Peak temperature between instrument jaws is significantly reduced with perivascular tissue in situ. This may result in a negative impact on sealing quality.

P02.19.A ASSESSMENT OF HEAT SHOCK PROTEIN HSP70 EXPRESSION IN THE GLIOBLASTOMA STEM-LIKE CELLS

Abstract BACKGROUND Glioblastoma multiforme (GBM) is a malignant brain tumor characterized by poor prognosis. The search of biomarkers for early diagnosis and as targets and for tumor theranostics due to the aggressiveness of the disease is urgently needed. One of the promising targets for therapy of brain tumors is 70 kDa heat shock protein (Hsp70), which has been found on the plasma membrane of various tumor cells (including GBM) but not on normal cells. GBM, like other types of tumors, has cellular heterogeneity. In particular, it contains tumor stem-like cells (TSLCs), which are considered analogues of healthy stem cells and are largely responsible for disease progression, recurrence and mediate resistance to ongoing therapy. In the present work, the histological sections of human GBM were analyzed to identify and quantify the colocalization of the heat shock protein Hsp70 with TSLC markers using the Opal Multiplex analysis system. MATERIAL AND METHODS Histological sections were prepared in accordance with the standard protocol using postoperative material from patients with newly diagnosed GBM (n=15). Zones of necrosis and living tissue were identified on tumor sections. Based on a phenotyping map using the Pathology Views™ fluorescence image analysis software, Hsp70 colocalization was quantified with Nestin and Sox2 in the areas of necrosis and viable tissue employing primary antibodies (anti-Hsp70, anti-Nestin and anti-Sox2). The preparations were stained using an Opal 3-Plex Manual Detection Kit according to the manufacturer's protocol. RESULTS The expression of Hsp70 and TSLC markers (Nestin and Sox2) was detected. The obtained quantitative data did not reveal significant differences in the distribution of cells expressing Hsp70 in the areas of necrosis and viable tissue (36 and 41%, respectively), which may indicate their uniform distribution in the tumor. Sufficiently high colocalization of Hsp70 with Nestin and Sox2 factors was detected mainly in the zone of viable tissue (57 and 30%, respectively). CONCLUSION Colocalization of Hsp70 with TSLC markers indicates the prospects for further studies of Hsp70, as well as its use for targeted therapy of malignant brain tumors.

Quantitative diffusion-weighted MRI response assessment in rhabdomyosarcoma: an international retrospective study on behalf of the European paediatric Soft tissue sarcoma Study Group Imaging Committee

ObjectiveTo investigate the feasibility of diffusion-weighted magnetic resonance imaging (DW-MRI) as a predictive imaging marker after neoadjuvant chemotherapy in patients with rhabdomyosarcoma.Material and methodsWe performed a multicenter retrospective study including pediatric, adolescent and young adult patients with rhabdomyosarcoma, Intergroup Rhabdomyosarcoma Study group III/IV, treated according to the European paediatric Soft tissue sarcoma Study Group (EpSSG) RMS2005 or MTS2008 studies. DW-MRI was performed according to institutional protocols. We performed two-dimensional single-slice tumor delineation. Areas of necrosis or hemorrhage were delineated to be excluded in the primary analysis. Mean, median and 5th and 95th apparent diffusion coefficient (ADC) were extracted.ResultsOf 134 included patients, 82 had measurable tumor at diagnosis and response and DW-MRI scans of adequate quality and were included in the analysis. Technical heterogeneity in scan acquisition protocols and scanners was observed. Mean ADC at diagnosis was 1.1 (95% confidence interval [CI]: 1.1–1.2) (all ADC expressed in * 10−3 mm2/s), versus 1.6 (1.5–1.6) at response assessment. The 5th percentile ADC was 0.8 (0.7–0.9) at diagnosis and 1.1 (1.0–1.2) at response. Absolute change in mean ADC after neoadjuvant chemotherapy was 0.4 (0.3–0.5). Exploratory analyses for association between ADC and clinical parameters showed a significant difference in mean ADC at diagnosis for alveolar versus embryonal histology. Landmark analysis at nine weeks after the date of diagnosis showed no significant association (hazard ratio 1.3 [0.6–3.2]) between the mean ADC change and event-free survival.ConclusionA significant change in the 5th percentile and the mean ADC after chemotherapy was observed. Strong heterogeneity was identified in DW-MRI acquisition protocols between centers and in individual patients.Graphical

Metastasis of solitary fibrous tumor to the parotid gland diagnosed on aspiration cytology supplemented with immunocytochemistry.

Solitary fibrous tumor (SFT) can occur in varied anatomic locations, most commonly being the extremities-superficial and deep soft tissues, followed by retroperitoneum, visceral organs, and the bone. Head and neck region may harbor a few with a predilection for the orbit and sinonasal tract, oral cavity, and rarely the salivary glands. The parotid gland is most involved among all salivary glands. Overall, SFT in salivary glands is rare and mostly seen as a primary tumor. The occurrence of metastasis to the parotid gland is exceedingly uncommon. We present cytological findings of SFT, metastatic to the parotid gland in a young 31-year-old lady where the diagnosis was confirmed with the help of immunocytochemistry (ICC) on the cell block. The smears were highly cellular and showed predominantly discrete round to oval tumor cells with mild pleomorphism, coarse chromatin, inconspicuous nucleoli, and scanty cytoplasm. Mitosis and focal areas of necrosis were noted. Lymphoglandular bodies were absent ruling out a non-Hodgkin lymphoma. Features were of a poorly differentiated malignant tumor with differentials being sarcoma, myoepithelial carcinoma, and carcinoma ex-pleomorphic adenoma. A panel of ICC was done and positivity for signal transducer and activator of transcription 6 (STAT6) helped in clinching the correct diagnosis of SFT.

The protection of ash trees against ash dieback by tree injections

Ash dieback caused by a non-native pathogenic fungus Hymenoscyphus fraxineus has been decimating populations of European Fraxinus species for over 30 years. Nevertheless, there is still a considerable amount of valuable ashes of this species in European cities and landscapes. Although it has been shown in many studies that the effect of H. fraxineus on adult and senescent trees tends to be rather chronic, the infection by this fungus also causes a substantial decrease in the lifespan of these trees. This research is dealing with the control of ashes using the tree injection technology. It is focused on the development of bark necroses, as this symptom results in the highest damage to a tree. Trees were first inoculated with two different strains of H. fraxineus. Trees with proven positive inoculation were injected with six different fungicides. Subsequently, over the course of nine months, the area of developing bark necroses was measured using image analysis methods. After the field part of the experiment was finished, the extent of the infection by H. fraxineus in tissues was determined and confirmed by PCR. Three out of six tested variants (Tr. 1: 7.1% propiconazole, Tr. 3: 7% propamocarb and 2.7% fosetyl, and Tr. 6: 0.015% sodium selenite) displayed significant deceleration of the growth of bark necroses compared to control. Tr. 3 demonstrated the highest effectiveness with only a very small increment of the necrosis area during the first three months after the injection treatment and a follow-up complete cessation of growth. Reisolations confirmed high mortality of H. fraxineus after this treatment. Hence the presented study can contribute to substantial prolongation of the lifespan of ashes in Europe.

Primary pancreatic Ewing sarcoma with metastases on FDG PET/CT

BackgroundEwing sarcoma (ES) is a highly aggressive malignant tumor most commonly affecting long bones. Extraskeletal Ewing sarcoma (EES) is a malignant tumor with aggressive behavior carrying bad prognosis with pancreas being an extremely rare primary site. We present a case of histopathologically proven EES of the pancreas in a young female who presented with abdominal pain. 18F-fluorodeoxyglucose positron emission tomography (18F FDG PET/CT) is a useful modality for detecting distant metastases in EES. It helps in diagnosis, localizing the primary, its extension, optimal treatment planning and evaluation of response to standard treatments available.Case presentationAn 18-year-old female presented with complaints of progressive abdominal pain and distention since 6 weeks. Physical examination was suggestive of a solid large mass in the upper left abdomen and decreased breath sounds with dullness in the left lower lung fields. On Contrast enhanced computed tomography (CECT) imaging, a large heterogeneously enhancing mass was seen arising from pancreas along with retroperitoneal lymphadenopathy. A moderate sized left sided pleural effusion with atelectasis of lower lobe of left lung was also noted. Histopathological analysis was suggestive of pancreatic ES following which the patient underwent five cycles of chemotherapy. Following this, she underwent 18F FDG PET/CT which showed hypermetabolic large mass arising from body and tail of pancreas with areas of internal necrosis along with left adrenal metastasis, retroperitoneal lymphadenopathy, a massive left pleural effusion and compressive atelectasis of left lower lobe. The patient expired within a week following PET/CT.ConclusionsEES most often presents in the late stage of the disease with vague symptoms. Timely diagnosis and initiation of treatment is of utmost importance considering the aggressiveness of the tumor. Establishing a diagnosis of Ewing sarcoma is especially difficult when the mass is arising from the pancreas. Imaging, histopathology and immunohistochemistry (IHC) play a key role in accurate diagnosis of such masses. 18F FDG PET/CT can be useful for detecting local and distant spread, operability, treatment planning and evaluation of response to chemotherapy.

Comparison of Gross Pathology between Classical and Recombinant Lumpy Skin Disease Viruses.

The pathology caused by three different isolates of lumpy skin disease virus, classical field cluster 1.2 strain Dagestan/2015, recombinant vaccine-like cluster 2.1 strain Saratov/2017, and cluster 2.2 strain Udmurtiya/2019, in cattle was compared from experimental infections. The infection of cattle was performed using intravenous administration of 2 mL of 105 TCID50/mL of each specific LSDV. Both classical and recombinant forms of LSDV cause pathological changes in the skin and lymph nodes, as well as the trachea and lungs. Due to circulatory disorders in the affected organs, multiple areas of tissue necrosis were observed, which, with the resurgence of secondary microflora, led to the development of purulent inflammation. Observed pathological changes caused by the recombinant vaccine-like strain Udmurtiya/2019 were characterized by a more pronounced manifestation of the pathoanatomical picture compared to the classical field strains Dagestan/2015 and Saratov/2017. Interestingly, Dagestan/2015 and Udmurtiya/2019 caused damage to the lymph nodes, characterized by serous inflammation and focal purulent lymphadenitis caused by purulent microflora. "Saratov/2017" did not cause pathology in the lymph nodes. All LSDVs were virulent and caused pathology, which was not distinguishable between viruses. This data set will serve as the experimentally validated basis for the comparative examination of novel LSDV strains in gross pathology.

Intratumoral heterogeneity after targeted therapy in murine cancer models with differing degrees of malignancy

IntroductionConventional morphologic and volumetric assessment of treatment response is not suitable for adequately assessing responses to targeted cancer therapy. The aim of this study was to evaluate changes in tumor composition after targeted therapy in murine models of breast cancer with differing degrees of malignancy via non-invasive magnetic resonance imaging (MRI). Materials and methodsMice bearing highly malignant 4T1 tumors or low malignant 67NR tumors were treated with either a combination of two immune checkpoint inhibitors (ICI, anti-PD1 and anti-CTLA-4) or the multi-tyrosine kinase inhibitor sorafenib, following experiments with macrophage-depleting clodronate-loaded liposomes and vessel-stabilizing angiopoietin-1. Mice were imaged on a 9.4 T small animal MRI system with a multiparametric (mp) protocol, comprising T1 and T2 mapping and diffusion-weighted imaging. Tumors were analyzed ex vivo with histology. Results and discussionsAll treatments led to an increase in non-viable areas, but therapy-induced intratumoral changes differed between the two tumor models and the different targeted treatments. While ICI treatment led to intratumoral hemorrhage, sorafenib treatment mainly induced intratumoral necrosis. Treated 4T1 tumors showed increasing and extensive areas of necrosis, in comparison to 67NR tumors with only small, but also increasing, necrotic areas. After either of the applied treatments, intratumoral heterogeneity, was increased in both tumor models, and confirmed ex vivo by histology. Apparent diffusion coefficient with subsequent histogram analysis proved to be the most sensitive MRI sequence. In conclusion, mp MRI enables to assess dedicated therapy-related intratumoral changes and may serve as a biomarker for treatment response assessment.

Morphological and Molecular Biological Features of the Systemic Inflammatory Response in Old Wistar Rats with High and Low Resistance to Hypoxia.

Morphological and molecular biological features of LPS-induced systemic inflammatory response were assessed in old male Wistar rats with high (HR) and low (LR) resistance to hypoxia. The systemic inflammatory response was modeled by intraperitoneal injection of E. coli O26:B6 LPS; the animals were sacrificed after 6 h. In histological sections, the number of neutrophils in the interalveolar septa and the area of necrosis in the liver were determined. The expression levels of Hif1a, Hif2a, Nfkb, Vegf, Il1b, Il6, Il10, and Tgfb mRNA in the liver and serum concentrations of HIF-1α and IL-1β were determined. In 4-6 h after LPS injection, 3 (43%) of 7 HR rats died, whereas no deaths were observed among LR rats. At 6 h after LPS injection, the number of neutrophils in the interalveolar septa of the lungs in LR rats was significantly higher than in HR rats, while the area of necrosis in the liver did not differ. At the same time, the mRNA expression levels of the proinflammatory cytokine genes Il1b and Il6 increased in the liver of both HR and LR rats, whereas the expression of Il10 increased only in HR rats. The expression of the Hif1a gene in the liver was higher in LR rats, but the content of HIF-1α protein in blood serum was higher in HR animals. These data should be taken into account when developing new approaches to the therapy of systemic inflammatory response in infectious and inflammatory diseases in the elderly.

Epithelioid extragastrointestinal stromal tumor in the pelvic cavity: A rare case at a rare location

ABSTRACT Extraintestinal gastrointestinal stromal tumors (EGISTs) are rare stromal tumors involving mesentery, omentum, and retroperitoneum. EGISTs of the pelvic cavity are extremely uncommon, and till date, no case of pure epithelioid type arising from the pelvic cavity has been reported. They pose a diagnostic pitfall as they may mimic ovarian malignancy. We present a unique case of an epithelioid extraintestinal stromal tumor arising from the pelvic cavity. A 43-43-year-old woman presented with complaints of abdominal pain and menstrual irregularities. Ultrasound examination revealed a large well-defined hypoechoic mass in the right pelvic cavity with a possible site of origin from the right ovary. A total hysterectomy with bilateral salpingo-oophorectomy was done. Histopathological examination revealed an encapsulated tumor comprising predominantly of nests and lobules of epithelioid cells with round-to-oval nuclei, vesicular chromatin, and moderate amount of eosinophilic to clear cytoplasm. Mitotic activity was brisk along with focal areas of necrosis. On immunohistochemistry, tumor cells were diffusely and strongly positive for DOG1 and CD117. They were negative for smooth muscle actin, vimentin, S-100, and HMB-45. Based on these findings, a final diagnosis of the high-grade GIST epithelioid variant was made. It is crucial to make the correct preoperative diagnosis of GIST, since these patients may benefit from neoadjuvant imatinib, especially in case of large tumor size.

246. SPATIAL HETEROGENEITY OF PD-L1 EXPRESSION INFLUENCE ITS ASSESSMENT IN ESOPHAGEAL SQUAMOUS CARCINOMA (ESCC)

Abstract Background Anti-PD-1/PD-L1 immunotherapy is a promising treatment for ESCC. High PD-L1 expression is a theoretical predictive biomarker, but the poor response is also observed in some patients with high PD-L1 expression. Biopsies assessment of PD-L1 expression always focuses on a single region which may be influenced by spatial heterogeneity of tumor microenvironment. This study investigates whether the PD-L1 expression by biopsy assessment in the different spatial regions is related to its paradoxical predictive value. Methods We prospectively included 30 consecutive patients with treatment-naïve, stage II-III ESCC who underwent upfront esophagectomy. Multi-region sampling was taken with endoscopic biopsy forceps (Fig. A) from four regions (A/B/C/D) from the proximal, distal, midpoint of tumor surface, and internal center, respectively. Each region was spaced ≥1 cm apart and avoided areas of severe necrosis. We compared the PD-L1 expression at different endoscopic-biopsy regions and the overall surgical-resected tumor specimen. PD-L1 expression was evaluated by immunohistochemistry using combined positive score (CPS). PD-L1 expression positive and negative is defined as CPS ≥ 1 and CPS &amp;lt; 1, respectively. Results Based on surgical-resected tumor, PD-L1 positive and high were 66.7% and 10% of 30 patients, respectively. PD-L1 positive/negative discordance rate of four sampling regions was 66.7% with poor concordance (kappa = 0.206) (Fig. B). Correlations between four biopsy regions of PD-L1 expression (Fig. C) revealed high in B and D (r = 0.77) but relatively low in others. Combining multi-region sampling for evaluation improved the correlation consistency of PD-L1 expression assessment (Fig. D). Comparing PD-L1 expression based on the endoscopic-biopsy region with the gold standard surgical-resected specimen, the sensitivity was increased by 12.5% for multi-region versus single-region assessment. Conclusion PD-L1 expression was spatially heterogeneous in ESCC, and a single region cannot reflect the PD-L1 status of the tumor, consistent with the observation of suboptimal prediction of PD-L1 expression for ESCC immunotherapy. Multi-region biopsy could help improve the assessment bias of PD-L1 expression related to spatial heterogeneity, and may help to improve the identification of the candidate for immunotherapy.

Reducing the Incidence of Post – Myocardial Infarction Heart Failure in an Era of Improved Primary Percutaneous Coronary Intervention through Novel Therapeutics (Protein Kinase C Beta II Inhibitor)

Heart failure (HF) complicating myocardial infarction (MI) remains a major source of morbidity, mortality and a huge health-care burden in the world. The incidence of symptomatic HF after primary percutaneous coronary intervention (PPCI) has been widely reported. PPCI has improved early survival of MI, but its impact on downstream HF is deliberated. Infarct size is the major determinant of the development of HF and prognosis. Efforts to reduce the size will therefore have a significant impact on the incidence of HF after PPCI. Resuscitation of the ischemic myocardium, heavily rests on timely reperfusion. However, cardiomyocyte death still occurs due to ischemic/reperfusion (I/R) injury mediated by oxidative stress. This phenomenon is known to largely contribute to the size of infarct. Activation by protein kinase C-βII (PKCβII) is attributed to the generation of reactive oxygen species (ROS) during this process and inhibition of this drive could salvage the myocardial tissue and subsequently preserve cardiac function. We hypothesized that myristic acid (Myr) and trans-activator of transcription (Tat) conjugated PKCβII inhibitor (Myr-Tat-PKCβII-) will exhibit robust cardioprotective effects in an in vivo porcine model, reflective of our ex vivo rat hearts model. Male Yorkshire pigs underwent regional I/R by occluding the second branch of the left anterior descending artery (LAD). This was achieved in the catheterization laboratory guided by fluoroscopy. A bolus of Myr-Tat-PKCβII- or scrambled control was infused into the LAD during reperfusion. Cardiac function was evaluated by an echocardiogram during the entire procedure, and assessed as the relative change at the end of 3hr reperfusion compared to baseline. Post-reperfused hearts were stained with Evans Blue dye to identify the area a risk and 1% triphenyltetrazolium chloride to delineate the area of necrosis. Infarct size and ejection fraction (EF) were analyzed via Student’s t- test. From mean baseline EF (61.4±0.5%), Myr-Tat-PKCβII- preserved EF with a relative change of 1.2±2.8% compared to 8.9±2.2% in control hearts (p&lt;0.05) at the end of reperfusion. Myr-Tat-PKCβII- reduced infarct size to 13.5±3.9% (n=4) compared to scrambled control hearts (27.5 ±7.9%, n=6). These results suggest Myr-Tat-PKCβII- significantly preserve EF after PPCI and reduce infarct size when given at reperfusion onset. This study gives a solid premise for future examination of Myr-Tat-PKCβII- in an 8 week in-vivo, porcine MI survival study.

Epithelioid dedifferentiated liposarcoma: A clinicopathological and molecular study of 6 cases

In this study, we present six cases of epithelioid dedifferentiated liposarcoma to further characterize its clinical and pathological features. The patients are all adult men with age at presentation ranging from 46 to 64 years (median 58.5 years). The patients presented with nonspecific symptoms of retroperitoneal mass, intermittent dull pain or discomfort. None of the patients had any prior history of a primary tumor. Radiological examinations revealed the presence of ill-demarcated heterogenous mass located in the deep soft tissue, including retroperitoneum (4 cases), pelvis and trunk (1 case each). Grossly, they appeared as solid tumors with focal areas of necrosis being presented in 2 cases. Histologically, all tumors were characterized by sheets of epithelioid cells that displayed marked cellular atypia and brisk mitotic activity. Variable portion of atypical lipomatous tumor/well-differentiated liposarcoma was present in 3 cases. By immunohistochemistry, the high-grade epithelioid component in all 6 cases showed strong and diffuse nuclear staining of MDM2, CDK4 and P16, with partial expression of AE1/AE3 in 3 cases. Fluorescence in situ hybridization analysis revealed high-level amplification of MDM2 in all 6 cases, with co-amplification of CDK4 in 3 cases. Follow up information showed that two patients died of the disease within one year despite multidisciplinary treatment. Due to the striking epithelioid appearance, this rare variant of dedifferentiated liposarcoma may be confused with undifferentiated epithelioid sarcoma, poorly differentiated carcinoma, mesothelioma or other malignancies with epithelioid phenotype. The study presented herein further highlights the aggressive clinical behavior of this unique tumor type. For patients with advanced disease, CDK4 inhibitor may provide an optional targeted treatment.

Composition and hepatoprotective effect of geopropolis of Melipona subnitida

Background: Geopropolis produced by bees of the subfamily Meliponinae (Hymenoptera, Apidae) is used in Brazilian popular medicine for the treatment of different diseases. The aim of the present work was to evaluate the hepatoprotective potential of the hydroethanolic extract from geopropolis (HPE) produced by Melipona subnitida Ducke. Additionally, the chemical composition of HPE and antioxidant activity were also evaluated. Methods: The hepatoprotective potential of HPE was evaluated by serum biochemical profile and macroscopic and histological examination of the liver of 24 Wistar rats (Rattus norvegicus), distributed in four experimental groups: G1 (negative control), G2 (HPE), G3 (HPE plus acetaminophen), G4 (acetaminophen - positive control). The HPLC-DADESI-MS/MS method was used to evaluate the chemical composition of HPE and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method was used to measure the antioxidant activity of geopropolis. Results: While HPE alone (G2) caused no liver injury, the livers of G4 exhibited color and histological alterations, extensive hemorrhagic suffusions, friable consistence, yellowish-white multifocal areas, degeneration and diffuse hepatocyte necrosis, in addition to altered levels of urea and enzymes TGO (glutamic-oxaloacetic transaminase) and TGP (glutamic pyruvic transaminase). HPE was shown to exert hepatoprotective effect in G3 animals, only mild hemorrhage and focal areas of vacuolar degeneration and hepatocyte necrosis having been observed. The geopropolis analyzed contains a high diversity of phenolic compounds, including chalcones, flavones and flavonols. high antioxidant activity was determined (IC50=48 μg/mL). Conclusions: The hydroethanolic extract of the geopropolis produced by M. subnitida has a high diversity of phenolic compounds and a high antioxidant activity. It has hepatoprotective action, as evidenced by a reduction in the levels of enzymaraes and the severity of the macroscopic and histological changes in livers of Wistar rats subjected to acetaminophen induced liver changes.

Nanoparticles targeting Sialyl-Tn for efficient tyrosine kinase inhibitor delivery in gastric cancer

Gastric cancer (GC) is the fourth leading cause of cancer-related deaths worldwide and, therefore, it is urgent to develop new and more efficient therapeutic approaches. Foretinib (FRT) is an oral multikinase inhibitor targeting MET (hepatocyte growth factor receptor) and RON (recepteur d'origine nantais) receptor tyrosine kinases (RTKs) that has been used in clinical trials for several solid tumors. Targeted uptake of therapeutic polymeric nanoparticles (NPs) represents a powerful approach in cancer cell drug delivery. Previously, a nanodelivery system composed of polymeric NPs functionalized with B72.3 antibody, which targets the tumor-associated antigen Sialyl-Tn (STn), has been developed. Herein, these NPs were loaded with FRT to evaluate its capacity in delivering the drug to multicellular tumors spheroids (MCTS) and mouse models. The data indicated that B72.3 functionalized FRT-loaded PLGA-PEG-COOH NPs (NFB72.3) specifically target gastric MCTS expressing the STn glycan (MKN45 SimpleCell (SC) cells), leading to a decrease in phospho-RTKs activation and reduced cell viability. In vivo evaluation using MKN45 SC xenograft mice revealed that NFB72.3 were able to decrease tumor growth, reduce cell proliferation and tumor necrosis. NFB72.3-treated tumors also showed inactivation of phospho-MET and phospho-RON. This study demonstrates the value of using NPs targeting STn for FRT delivery, highlighting its potential as a therapeutic application in GC. Statement of significanceDespite the advances in gastric cancer therapeutics, it remains one of the diseases with the highest incidence and mortality in the world. Combining targeted therapies with a controlled drug release is an attractive strategy to reduce drug cytotoxic effects and improve specific drug delivery efficiency to the cancer cells. Thus, we developed nanoparticles loaded with a tyrosine kinase inhibitor and targeting a specific tumor glycan exclusive of cancer cells. In in vivo gastric cancer xenograft mice models, these nanoparticles efficiently reduced tumor growth, cell proliferation and tumor necrosis area and inactivated phosphorylation of targeting receptors. This approach represents an innovative therapeutic strategy with high impact in gastric cancer.

Morphological and molecular-biological features of glioblastoma progression in tolerant and susceptible to hypoxia Wistar rats

Hypoxia is a major pathogenetic factor in many cancers. Individual resistance to suboptimal oxygen availability is subject to broad variation and its possible role in tumorigenesis remains underexplored. This study aimed at specific characterization of glioblastoma progression in male tolerant and susceptible to hypoxia Wistar rats. Hypoxia resistance was assessed by gasping time measurement in an 11,500 m altitude-equivalent hypobaric decompression chamber. Based on the outcome, the animals were assigned to three groups termed ‘tolerant to hypoxia’ (n = 13), ‘normal’, and ‘susceptible to hypoxia’ (n = 24). The ‘normal’ group was excluded from subsequent experiments. One month later, the animals underwent inoculation with rat glioblastoma 101.8 followed by monitoring of survival, body weight dynamics and neurological symptoms. The animals were sacrificed on post-inoculation days 11 (subgroup 1) and 15 (subgroup 2). Relative vessels number, necrosis areas and Ki-67 index were assessed microscopically; tumor volumes were determined by 3D reconstruction from histological images; serum levels of HIF-1α, IL-1β, and TNFα were determined by ELISA. None of the tolerant to hypoxia animals died of the disease during observation period, cf. 85% survival on day 11 and 55% survival on day 15 in the susceptible group. On day 11, proliferative activity of the tumors in the tolerant animals was higher compared with the susceptible group. On day 15, proliferative activity, necrosis area and volume of the tumors in the tolerant to hypoxia animals were higher compared with the susceptible group. ELISA revealed no dynamics in TNFα levels, elevated levels of IL-1β in the susceptible animals on day 15 in comparison with day 11 and tolerant ones. Moreover, there were elevated levels of HIF-1α in the tolerant animals on day 15 in comparison with day 11. Thus, the proliferative activity of glioblastoma cells and the content of HIF-1α were higher in tolerant to hypoxia rats, but the mortality associated with the tumor process and IL-1β level in them were lower than in susceptible animals. Specific features of glioblastoma 101.8 progression in tolerant and susceptible to hypoxia rats, including survival, tumor growth rates and IL-1β level, can become the basis of new personalized approaches for cancer diseases treatment in accordance to individual hypoxia resistance.

Necrotizing fasciitis after total abdominal hysterectomy: A case report.

Necrotizing fasciitis is rapidly progressive infection with high mortality rate. This study aimed to summarize the clinical and pathological presentation of this case. A 46-year-old woman patient presented to our emergency department of an obstetric and gynecology clinic on the 8th day after total abdominal hysterectomy. The abdominal wall showed pronounced signs of inflammation. Abundant purulent content was oozing from the abdominal wound. The patient underwent surgery. Areas of necrosis were observed on the skin around the wound, the subcutaneous fatty tissue was necrotic around the incision site, and the fascia was completely dehisced. Wound debridement and flap cutting of the anterior abdominal wall were performed. Metronidazole, ceftriaxone, and vancomycin were administered intravenously. A plastic surgeon suggested daily debridement and toileting of the wound in the operating room. Swabs of the abdominal cavity, abscess cavity, and abdominal wound were obtained, and Enterococcus faecalis was isolated. After the negativism of microbiological swabs, excochleation of granulation tissue was performed by a plastic surgeon. Nineteen days after the relaparotomy, the patient was discharged in good general condition with advice for further monitoring and therapy. Successful treatment of necrotizing fasciitis can be achieved through an initial diagnosis, adequate debridement, empirical broad-spectrum antibiotic coverage, and multidisciplinary treatment.

BSBM-07 HIGH-DIMENSIONAL HISTOPATHOLOGIC EVALUATION OF THE HYPOXIC MICROENVIRONMENT IN GLIOBLASTOMA

Abstract Rapidly growing solid tumors such as glioblastoma (GBM) are characteristically hypoxic, displaying large areas of necrosis surrounded by hyperproliferative pseudopalisading cells. Intra-tumoral hypoxia develops over time in the three-dimensional space and the degree of tissue oxygenation is a dynamic process that varies continuously. Combined with the extensive inter- and intra-tumoral heterogeneity associated with GBM at the bulk and single cell level, hypoxia contributes to a gradient of molecular alterations that are specific to the different cell populations that make up the bulk of the tumor and reside in specific niches. To date, high dimensional histopathologic analyses of the hypoxic regions within GBM tissue have not been performed. Here, we took a combined spatial and single-cell proteomic profiling approach to investigate the histopathologic features of hypoxia by leveraging a unique clinical study where the exogenous hypoxia marker pimonidazole (PIMO) is administered to patients with GBM prior to surgery. Tissue specimens were subjected to imaging mass cytometry and serial immunohistochemistry using a panel of 27 markers associated with cellular hallmarks of hypoxia, metabolism, proliferation, stemness, angiogenesis, and immune cell types. We took high-resolution imaging and statistical approaches to explore the interplay of the different markers within hypoxic regions of primary and recurrent GBMs, in addition to IDH-mutant gliomas. Our findings elucidate the expression pattern of key biological markers relative to one another, altered composition of different cell types, along with differential proliferative, transcriptional, and translational activation states associated with each cell type within the hypoxic regions of GBM.

Small extracellular vesicles delivering lncRNA WAC-AS1 aggravate renal allograft ischemia‒reperfusion injury by inducing ferroptosis propagation.

Ferroptosis is a predominant contributor to renal ischemia reperfusion injury (IRI) after kidney transplant, evoking delayed graft function and poorer long-term outcomes. The wide propagation of ferroptosis among cell populations in a wave-like manner, developing the "wave of ferroptosis" causes a larger area of tubular necrosis and accordingly aggravates renal allograft IRI. In this study, we decipher a whole new metabolic mechanism underlying ferroptosis and propose a novel spreading pathway of the "wave of ferroptosis" in the renal tissue microenvironment, in which renal IRI cell-secreted small extracellular vesicles (IRI-sEVs) delivering lncRNA WAC-AS1 reprogram glucose metabolism in adjacent renal tubular epithelial cell populations by inducing GFPT1 expression and increasing hexosamine biosynthesis pathway (HBP) flux, and consequently enhances O-GlcNAcylation. Additionally, BACH2 O-GlcNAcylation at threonine 389 in renal tubular epithelial cells prominently inhibits its degradation by ubiquitination and promotes importin α5-mediated nuclear translocation. We present the first evidence that intranuclear BACH2 suppresses SLC7A11 and GPX4 transcription by binding to their proximal promoters and decreases cellular anti-peroxidation capability, accordingly facilitating ferroptosis. Inhibition of sEV biogenesis and secretion by GW4869 and knockout of lncRNA WAC-AS1 in IRI-sEVs both unequivocally diminished the "wave of ferroptosis" propagation and protected against renal allograft IRI. The functional and mechanistic regulation of IRI-sEVs was further corroborated in an allograft kidney transplant model and an in situ renal IRI model. In summary, these findings suggest that inhibiting sEV-mediated lncRNA WAC-AS1 secretion and targeting HBP metabolism-induced BACH2 O-GlcNAcylation in renal tubular epithelial cells may serve as new strategies for protecting against graft IRI after kidney transplant.