UNASSIGNED: The utilization of smartphone-assisted evaluation is emerging in the field of histopathology. This technique improves the adequacy of samples at the bedside, avoids procedure-related complications, reduces unnecessary repeat biopsies, and saves the cost of the procedure. This study aims to compare the number of glomeruli in a renal biopsy specimen obtained by an ultrasound-guided percutaneous needle biopsy, counted at the bedside using a smartphone fitted with a 16-megapixel macro lens (Bedside method) with that observed under a light microscope after the processing of the biopsy specimen (LM method).
UNASSIGNED: In this prospective cohort study, 24 consecutive adult patients (48 kidney biopsy samples) who underwent kidney biopsies were enrolled. All specimens were extracted by an ultrasound-guided percutaneous renal biopsy from the lower pole of the left kidney. Patients\' demographics and clinical data were prospectively collected. The number of glomeruli in all the biopsy specimens was counted using a smartphone fitted with a 16-megapixel macro lens at the bedside (Bedside method) and subsequently under a light microscope by a pathologist after processing the biopsy specimen (LM method). Seven or more glomeruli in the specimen were considered adequate in our study.
UNASSIGNED: The mean age of patients at biopsy was 46.9 ± 16 years with slightly male predominance (54.2%). A total of 47 specimens were obtained from 24 patients. Of the 24 patients, 22 had native kidney biopsy and 2 had renal allograft biopsy. The average number of cores obtained per patient was 1.96. The length of core specimens ranged from 1.5 to 2 cm. A good agreement was found between bedside adequacy and slide adequacy, κ =0.684, P = 0.000. The positive agreement rate and negative agreement rate were 91.4% and 23.1%, respectively.
UNASSIGNED: In the modern era of technology, the smartphone is a good tool to evaluate the adequacy of biopsy specimens at the bedside.

AKT is one of the overexpressed targets in nonsmall cell lung cancer (NSCLC) and plays an important role in its progression and offers an attractive target for the therapy. The PI3K/AKT/mTOR pathway is upregulated in NSCLC. Acridone is an important heterocycle compound which treats cancer through various mechanisms including AKT as a target. In the present work, the study was designed to evaluate the safety profile of three acridone derivatives (AC-2, AC-7, and AC-26) by acute and repeated dose oral toxicity. In addition to this, we also checked the pAKT overexpression and its control by these derivatives in tumor xenograft model. The results from acute and repeated dose toxicity showed these compounds to be highly safe and free from any toxicity, mortality, or significant alteration in body weight, food, and water intake in the rats. In the repeated dose toxicity, compounds showed negligible variations in a few hematological parameters at 400 mg/kg. The histopathology, biochemical, and urine parameters remained unchanged. The xenograft model study demonstrated AC-2 to be inhibiting HOP-62 induced tumor via reduction in p-AKT1 (Ser473) expression significantly. In immunofluorescence staining AC-2 treated tissue section showed 2.5 fold reduction in the expression of p-AKT1 (Ser473). Histopathology studies showed the destruction of tumor cells with increased necrosis after treatment. The study concluded that AC-2 causes cell necrosis in tumor cells via blocking the p-AKT1 expression. The findings may provide a strong basis for further clinical applications of acridone derivatives in NSCLC.

BACKGROUND: Malignant pleural effusion (MPE) is a frequent complication of advanced malignancies. In this pilot study, we characterized the immune landscapes of MPEs, compared them to their primary tumor (PT) samples from breast carcinoma (BC) and lung adenocarcinoma (LADC), and tested the utility of multiplexed image technology in cytological samples.
METHODS: We evaluated the immune contexture of 6 BC and 5 LADC MPEs and their PTs using 3 multiplex immunofluorescence panels. We explored the associations between sample characteristics and pleural effusion-free survival.
RESULTS: No MPE samples had positive programmed death-ligand 1 expression in malignant cells, although 3 of 11 PTs has positive programmed death-ligand 1 expression (more than 1% expression in malignant cells). Overall, in LADC samples, cluster of differentiation 3 (CD3)+ T cells and CD3+CD8+ cytotoxic T cells predominated (median percentages for MPEs versus PTs: 45.6% versus 40.7% and 4.7% versus 6.6%, respectively) compared with BC. CD68+ macrophages predominated in the BC samples (medians for MPEs 61.2% versus PTs for 57.1%) but not in the LADC samples. Generally in PTs, CD3+CD8+ forkhead box P3+ T cells and the median distances from the malignant cells to CD3+CD8+Ki67+ and CD3+ programmed cell death protein 1 + T cells correlated to earlier MPE after PT diagnosis.
CONCLUSIONS: The immune cell phenotypes in the MPEs and PTs were similar within each cancer type but different between BC versus LADC. An MPE analysis can potentially be used as a substitute for a PT analysis, but an expanded study of this topic is essential.

The regulation of female fertility in mammals depends on critical processes during oocyte development and maturation. Therefore, it is crucial to use specific approaches when studying mammalian female fertility to preserve ovary and oocyte structures effectively. The methods of collecting and culturing ovaries and oocytes play an essential role in the study of mammalian follicle development and oocyte quality. This chapter presents a collection of protocols that focus on various methods for studying mammalian ovaries and oocytes, providing researchers with a variety of approaches to choose from.

UNASSIGNED: The data referring to the value of direct immunofluorescence on formalin-fixed, paraffin-embedded tissue (IF-Paraffin) in the diagnosis of renal diseases is controversial. The aim of this study was to investigate whether renal biopsies evaluated by routine immunofluorescence on frozen tissue (IF-Frozen) would yield adequate findings to confirm diagnoses when the IF-Paraffin technique was applied.
UNASSIGNED: To show immunoglobulins, complement components, and light chains, 55 native renal biopsies were subjected to IF-Paraffin and IF-Frozen staining techniques. The intensity of the staining was compared, and the sensitivity and specificity were calculated.
UNASSIGNED: The IF-Paraffin technique showed a sensitivity of 89%, 81%, 86%, 30%, 71%, 60%, and 77% for IgG, IgM, IgA, C1q, C3, κ, and λ, respectively, whereas specificity was 91%, 100%, 100%, 96%, 94%, 98%, and 100%. It showed diagnostic findings in 87% of cases. Compared to cases that had both IF-Paraffin and IF-Frozen staining techniques, 43 of 55 showed either equal intensity for the diagnostic immunoglobulin/complement or a little difference.
UNASSIGNED: Direct immunofluorescence on formalin-fixed, paraffin-embedded sections cannot replace immunofluorescence on frozen sections in the assessment of renal biopsies, but may be a \"salvage technique\" when frozen tissue is insufficient or unavailable and must be interpreted with great caution.

BACKGROUND: Endothelial cells (ECs) play a major role in malaria pathogenesis, as a point of direct contact of parasitized red blood cells to the blood vessel wall. The study of cytoskeleton structures of ECs, whose main functions are to maintain shape and provide strength to the EC membrane is important in determining the severe sequelae of Plasmodium falciparum malaria. The work investigated the cytoskeletal changes (microfilaments-actin, microtubules-tubulin and intermediate filaments-vimentin) in ECs induced by malaria sera (Plasmodium vivax, uncomplicated P. falciparum and complicated P. falciparum), in relation to the levels of pro-inflammatory cytokines.
METHODS: Morphology and fluorescence intensity of EC cytoskeleton stimulated with malaria sera were evaluated using immunofluorescence technique. Levels of tumour necrosis factor (TNF) and interferon (IFN)-gamma (γ) were determined using enzyme-linked immunosorbent assay (ELISA). Control experimental groups included ECs incubated with media alone and non-malaria patient sera. Experimental groups consisted of ECs incubated with malaria sera from P. vivax, uncomplicated P. falciparum and complicated P. falciparum. Morphological scores of cytoskeletal alterations and fluorescence intensity were compared across each experiment group, and correlated with TNF and IFN-γ.
RESULTS: The four morphological changes of cytoskeleton included (1) shrinkage of cytoskeleton and ECs with cortical condensation, (2) appearance of eccentric nuclei, (3) presence of \"spiking pattern\" of cytoskeleton and EC membrane, and (4) fragmentation and discontinuity of cytoskeleton and ECs. Significant damages were noted in actin filaments compared to tubulin and vimentin filaments in ECs stimulated with sera from complicated P. falciparum malaria. Morphological damages to cytoskeleton was positively correlated with fluorescence intensity and the levels of TNF and IFN-γ.
CONCLUSIONS: ECs stimulated with sera from complicated P. falciparum malaria showed cytoskeletal alterations and increased in fluorescence intensity, which was associated with high levels of TNF and IFN-γ. Cytoskeletal changes of ECs incubated with complicated P. falciparum malaria sera can lead to EC junctional alteration and permeability changes, which is mediated through apoptotic pathway. The findings can serve as a basis to explore measures to strengthen EC cytoskeleton and alleviate severe malaria complications such as pulmonary oedema and cerebral malaria. In addition, immunofluorescence intensity of cytoskeleton could be investigated as potential prognostic indicator for malaria severity.

In this study, bile acid-based vesicles and nanoparticles (i.e., bilosomes and biloparticles) are studied to improve the water solubility of lipophilic drugs. Ursodeoxycholic acid, sodium cholate, sodium taurocholate and budesonide were used as bile acids and model drugs, respectively. Bilosomes and biloparticles were prepared following standard protocols with minor changes, after a preformulation study. The obtained systems showed good encapsulation efficiency and dimensional stability. Particularly, for biloparticles, the increase in encapsulation efficiency followed the order ursodeoxycholic acid < sodium cholate < sodium taurocholate. The in vitro release of budesonide from both bilosytems was performed by means of dialysis using either a nylon membrane or a portion of Wistar rat small intestine and two receiving solutions (i.e., simulated gastric and intestinal fluids). Both in gastric and intestinal fluid, budesonide was released from bilosystems more slowly than the reference solution, while biloparticles showed a significant improvement in the passage of budesonide into aqueous solution. Immunofluorescence experiments indicated that ursodeoxycholic acid bilosomes containing budesonide are effective in reducing the inflammatory response induced by glucose oxidase stimuli and counteract ox-inflammatory damage within intestinal cells.

UNASSIGNED: Despite advances in diagnostics and therapeutics, two-thirds of oral cancer patients present with advanced disease, which increases both the morbidity and mortality risk. Circulating tumour cells (CTCs) are released in the circulation by primary tumours and have been demonstrated to have significant correlations between their occurrence and disease progression.
UNASSIGNED: To characterize the circulating tumour cells in subjects with histologically diagnosed oral squamous cell carcinoma (OSCC).
UNASSIGNED: This pilot study was undertaken with ten fresh blood samples (6 ml each). Five samples from apparently healthy individuals and five OSCC samples were cultured and subjected to flow cytometric analysis for CD44 expression. Immunostaining was done using CD44 and EpCAM markers.
UNASSIGNED: Several cells in OSCC samples showed EpCAM and CD44 positivity following immunostaining. However, flow cytometry performed with CD44 alone was not specific for OSCC samples. Hence, proving that CD44 and EpCAM when used in conjunction can help to characterize CTCs.
UNASSIGNED: The findings of our study suggest that the demonstration of CTCs is feasible and helps in understanding of disease progression and metastatic risk. Sensitive detection of CTCs from blood samples can serve as an implicit tool in early cancer diagnosis and prognosis through liquid biopsy which in itself is minimally invasive and time-saving.

This study aims to evaluate the responses of peripheral blood monocyte-derived macrophages (PBMDMs) from Theileria orientalis carrier cattle following exposure to Pasteruella multocida B:2 (PM B:2) and latex beads. Twenty-six male crossbred Kedah-Kelantan (KK) cattle were sampled for this study and quantitative PCR (qPCR) was employed in the detection of T. orientalis MPSP gene. Bactericidal assay using a 10:1 multiplicity of infection was performed to measure the phagocytosis and intracellular killing of PM B:2 by PBMDMs. The cell cultures were inoculated with 107 cfu/mL of PM B:2 and incubated in a humidified incubator. The absence of clinical signs, previous history of T. orientalis infection and an MPSP gene copy number below 15,000 GC/μL suggest that the cattle were asymptomatic chronic carriers. A non-significant phagocytic and mean cell death rates were observed in the PBMDMs of T. orientalis positive cattle relative to clinically healthy cattle (CHC) (p > 0.05). The PBMDMs of T. orientalis positive cattle had the lowest mean rate of intracellular killing relative to the CHC at the 30th minute post-infection only (p < 0.05). Exposure to latex beads caused an increase in the appearance of multinucleated macrophages following incubation of PBMDMs from T. orientalis positive cattle. Furthermore, the phagocytic index of PBMDMs of T. orientalis positive cattle were low or poor compared to that of CHC (p = 0.000). Therefore, our findings suggest that PBMDMs from cattle with chronic T. orientalis infection can efficiently phagocytise and kill PM: B2 but exhibited poor phagocytosis ability for foreign bodies despite appearance of multinucleated macrophages.

The inflammasome is a multiprotein complex that is responsible for mounting an innate immune response through the activation of caspase-1 and the cleavage of interleukin-1β. This multiprotein complex plays an important role in a variety of central nervous system (CNS) diseases and conditions such as Alzheimer\'s disease, Parkinson\'s disease, multiple sclerosis, stroke, and traumatic brain injury, among others. Here we describe methodological procedures to carry out immunoblotting and immunohistochemical techniques used to study inflammasome signaling in CNS tissues (brain and spinal cord).