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Oral cancer ranks as the 16th most prevalent form of cancer worldwide and is a significant concern in Southeast Asia, primarily due to the extensive use of tobacco. Chemokines contribute to a wide range of illness situations. This study aimed to investigate the immunohistochemistry expression of CCL28 and its receptor CCR10 in oral squamous cell carcinoma and examine their relationship with tumor progression. This study retrospectively examined tissues of oral squamous cell carcinoma that were preserved by formalin fixation and paraffin embedding. The study includes two groups: Group I included 50 cases of OSCC with varying histological grades, whereas Group II comprised 50 samples of normal oral mucosa (NOM) obtained from healthy individuals who did not engage in any tobacco use. The cytoplasmic placement of the study groups allowed us to determine the expression patterns of CCL28 and CCR10. The study found a strong link between CCL28 and CCR10 immunohistochemical expression in oral squamous cell carcinoma. There were big differences in the amount of CCR10 staining in the 96 cases that showed positive immunoexpression. This shows that these two molecules have a big impact on how OSCC acts biologically. Various stages of oral squamous cell carcinoma and healthy oral mucosa exhibited simultaneous expression of CCL28 and CCR10. In summary, we can utilize CCR10 and CCL28 co-expression, along with other biomarkers, to aid in the prognosis of oral squamous cell carcinoma (OSCC). [ABSTRACT FROM AUTHOR]

Background: Chronic wasting disease (CWD) is a fatal transmissible spongiform encephalopathy in cervids. The disease is caused by a pathogenic prion, namely PrPSc. Currently, diagnosis of CWD relies on IHC detection of PrPSc in the obex or retropharyngeal lymph nodes (RPLN) or ELISA screening of obex and RPLN followed by IHC confirmation of positive results. In this study, we assessed the performance characteristics of two immunoassays: CWD Ag-ELISA and TeSeE ELISA, RT-QuIC, and MEMS biosensor via testing 30 CWD + and 30 CWD- white-tailed deer RPLN samples. Results: Both CWD Ag-ELISA and TeSeE ELISA correctly identified all CWD + and CWD- samples. A greater intra-assay coefficient of variation (CV) in S/P ratios was observed for the TeSeE ELISA (16.52%), compared to CWD Ag-ELISA (9.49%). However, the high CV did not affect the qualitative results of triplicate assays when the corresponding manufacturer's cutoff was used. The MEMS biosensor not only correctly identified all CWD + and CWD- RPLN samples, but also demonstrated a 100% detection rate for all CWD + samples at dilutions from 10− 0 to 10− 3. Evaluation of RT-QuIC indicated that the rate of false negative reactions decreased from 21.98% at 10− 2 dilution to 0% at 10− 4 and 10− 5 dilutions; and the rate of false positive reactions reduced from 56.42% at 10− 2 dilution to 8.89% and 2.22% at 10− 4 and 10− 5 dilutions, respectively. Based on a stringent threshold of 2 x the first 10 fluorescent readings of each well and a final cutoff of 2/3 positive reactions for each sample, RT-QuIC correctly identified all positive and negative samples at 10− 4 and 10− 5 dilutions. Both MEMS biosensor and RT-QuIC achieved 100% sensitivity and 100% specificity under the experimental conditions described in this study. Conclusions: The two immunoassays (CWD Ag-ELISA and TeSeE ELISA) performed comparably on white-tailed deer RPLN samples. MEMS biosensor is a reliable portable tool for CWD diagnosis and RT-QuIC can be used for routine testing of CWD if appropriate testing parameters and interpretive criteria are applied. [ABSTRACT FROM AUTHOR]

For almost a decade predicting the response of malignant tumours to treatment with immunotherapies has been dependent on PD-L1 expression by immunohistochemistry. Much has been learned about the uniquely challenging application of PD-L1 immunohistochemistry which is utilized in a constantly expanding array of tumour types and clinical contexts. More recently, other predictive biomarkers have acquired clinical validation including tumour mismatch repair deficiency. In this short review, I will assess the strengths and weaknesses of PD-L1 immunohistochemistry, including pre-analytics, analytics and post-analytics in the context of the complex underlying biology. In doing so, I hope to provide some practical guidance that will help optimize its application in routine practice. I will also highlight the various contexts in which immunohistochemistry for PD-L1 and mis-match repair is used to predict response to immunotherapies and how ongoing developments of novel immunohistochemistry based biomarkers might assist in strengthening the ability to predict response to immunotherapies. [ABSTRACT FROM AUTHOR]

Simple Summary: Cyclin-dependent kinase inhibitor 2A/B (CDKN2A/B) loss is a key factor in diagnosing meningiomas as Central Nervous System (CNS) WHO grade 3 tumors. Typically, detecting this gene loss involves costly and complex techniques like sequencing or FISH. However, the MTAP gene, which is located near CDKN2A/B on the same chromosome, may provide a more accessible way to detect these losses through a simpler, more affordable test called immunohistochemistry (IHC). This study explored different concentrations and antibodies for MTAP and p16 across two institutions to evaluate their potential as surrogate markers for CDKN2A/B loss. The results showed that p16 expression varied and did not align with either MTAP expression or CDKN2A FISH results. This study suggests that IHC for MTAP is a promising, cost-effective method for identifying high-grade meningiomas, offering a quicker and less expensive alternative to existing techniques. Background: Homozygous cyclin-dependent kinase inhibitor 2A/B (CDKN2A/B) loss is one of the parameters that support the designation of meningiomas as Central Nervous System (CNS) WHO grade 3 tumors. Evaluation of CDKN2A/B by sequencing or Fluorescence in situ hybridization (FISH) is costly and not always readily accessible. An immunohistochemistry (IHC)-based marker for the evaluation of CDKN2A/B loss would provide faster results at a lower cost. Methods: This retrospective study included patients diagnosed with meningioma at our institution between 2016 and 2019. Archival tumor tissue was used for analysis. MTAP immunohistochemistry (IHC) was performed at various dilutions (1:1200, 1:400, 1:200, 1:100) using two different antibodies, and p16 IHC was conducted simultaneously. These analyses were carried out at two different institutions. To determine the sensitivity and specificity of MTAP and p16 as surrogate markers for CDKN2A/B loss, CDKN2A FISH was utilized as the gold standard. Results: Overall, 46/49 tumors showed strong MTAP staining (94%) at institution 1, and 44/49 (90%) showed either faint positive or positive results at institution 2. One grade 3 meningioma that demonstrated homozygous CDKN2A loss by FISH also showed loss of MTAP expression by IHC. One grade 2 meningioma showed regional CDKN2A loss by FISH and variable MTAP expression under different IHC conditions. MTAP expression evaluation was superior at a dilution of 1:100 with the Abnova Anti-MTAP Monoclonal antibody. Conclusions: P16 expression was variable and did not correlate with either MTAP expression or CDKN2A FISH results. MTAP IHC is a promising surrogate marker for the evaluation of CDKN2A status in meningiomas. [ABSTRACT FROM AUTHOR]

Abstract Background Chronic wasting disease (CWD) is a fatal transmissible spongiform encephalopathy in cervids. The disease is caused by a pathogenic prion, namely PrPSc. Currently, diagnosis of CWD relies on IHC detection of PrPSc in the obex or retropharyngeal lymph nodes (RPLN) or ELISA screening of obex and RPLN followed by IHC confirmation of positive results. In this study, we assessed the performance characteristics of two immunoassays: CWD Ag-ELISA and TeSeE ELISA, RT-QuIC, and MEMS biosensor via testing 30 CWD + and 30 CWD- white-tailed deer RPLN samples. Results Both CWD Ag-ELISA and TeSeE ELISA correctly identified all CWD + and CWD- samples. A greater intra-assay coefficient of variation (CV) in S/P ratios was observed for the TeSeE ELISA (16.52%), compared to CWD Ag-ELISA (9.49%). However, the high CV did not affect the qualitative results of triplicate assays when the corresponding manufacturer’s cutoff was used. The MEMS biosensor not only correctly identified all CWD + and CWD- RPLN samples, but also demonstrated a 100% detection rate for all CWD + samples at dilutions from 10− 0 to 10− 3. Evaluation of RT-QuIC indicated that the rate of false negative reactions decreased from 21.98% at 10− 2 dilution to 0% at 10− 4 and 10− 5 dilutions; and the rate of false positive reactions reduced from 56.42% at 10− 2 dilution to 8.89% and 2.22% at 10− 4 and 10− 5 dilutions, respectively. Based on a stringent threshold of 2 x the first 10 fluorescent readings of each well and a final cutoff of 2/3 positive reactions for each sample, RT-QuIC correctly identified all positive and negative samples at 10− 4 and 10− 5 dilutions. Both MEMS biosensor and RT-QuIC achieved 100% sensitivity and 100% specificity under the experimental conditions described in this study. Conclusions The two immunoassays (CWD Ag-ELISA and TeSeE ELISA) performed comparably on white-tailed deer RPLN samples. MEMS biosensor is a reliable portable tool for CWD diagnosis and RT-QuIC can be used for routine testing of CWD if appropriate testing parameters and interpretive criteria are applied.

Aim: Human epidermal growth factor receptor-2 (HER2) is a well-established prognostic and predictive biomarker. It is an FDA-approved therapeutic target for HER2 positive breast, gastroesophageal, and more recently, lung and colon cancers. It is an emerging biomarker in biliary tract, bladder, cervical, endometrial, ovarian, and pancreatic cancers. The emergence of new indications warrants further characterization of HER2 expression in diverse cancer populations. This study investigated HER2 expression in solid tumour samples and the feasibility of obtaining these results. Methods: Prospective consent was obtained at a Canadian tertiary academic cancer center from adult oncology patients who were referred for molecular genetic testing of malignant tissue samples. Standard HER2-targeted malignancies were considered breast and gastroesophageal, and were excluded from this study. Between July 2020 and November 2023, 499 samples of solid tumors underwent immunohistochemistry (IHC) HER2 staining. A median turnaround time (TAT) of 14 days would be considered feasible for clinical decision making. Results: The mean age (± SD) of participants was 67 ± 12.5 years, with 270 (54%) male and 229 (46%) female. HER2 protein expression was measured in 42 unique cancer types. IHC levels of 0, 1+, 2+, and 3+ were reported and were 43%, 12%, 35%, and 10% of all analyzable samples respectively (tissue inadequate in 3% of samples). The median TAT for HER2 expression results from time of request to result in release was 18 (interquartile range, 11 to 30) days. Conclusions: HER2 protein expression varies widely between different cancer types. TAT for HER2 IHC results was a median of 18 days, which is close to our feasibility cut-off.

Fixation inhibits autolysis and is an essential step in disease. Our work examined how neutral buffer NBF (NBF), zinc formalin (ZF), and Bouin's fluid (BF) affected tissue specific CD4, CD68, and Pax8 anti-genetic determinants. Fifty-one sections were analyzed for each fixative marker. One hundred fifty-three portions were scored 0, 1, 2, and 3. The comparison between scores in the detection of CD4 epitopes reflects that BF fluid was significant compared to NBF and ZF at a level of p< 0.05, whereas NBF comes after ZF. Surprisingly, ZF has no significant scores in the context of CD4 detection. On the other hand, CD68 detection at the score one level revealed that NBF and ZF showed significant CD68 anti-genetic determinants, whereas none of the CD68 epitopes were significantly determined in BF fluid. Eventually, both NBF and BF produced an excellent determination of Pax8 epitopes in contrast to ZF. However, there is no significant difference between them. Regarding scores three comparisons in all fixatives, BF fluid showed high-quality CD4 marker expression in contrast to NBF and ZF; however, neither CD68 nor Pax8 showed any significance in this context. We conclude that NBF showed an excellent quality result in detecting all epitopes. However, BF fluid was supreme in detecting anti-genetic determinants of CD4 and Pax8 epitopes in score 3. Additionally, ZF could distinguish CD68 and Pax8 but failed to detect CD4. Conversely, no significant differences were recorded between both B and NBF.

The establishment of immunotherapy applying immune checkpoint inhibitors (ICI) has provided an important new option for the treatment of solid malignant diseases. However, different tumor entities show dramatically different responses to this therapy. BCC responds worse to anti-PD-1 ICIs as compared to cSCC. Differential immune checkpoint expression could explain this discrepancy and, therefore, the aim of this study was to analyze activating and inhibitory immune checkpoints in cSCC and BCC tissues. Tissue microarrays of the invasive front as well as the tumor core of BCC and cSCC samples were used to evaluate PD-1, PD-L1, CD28, and CD86 expression and their topographic distribution profiles by chromogenic immunohistochemistry. QuPath was used to determine the labeling index. The expression of PD-1, PD-L1, and CD28 was significantly higher in both the tumor core and the invasive front of cSCC samples as compared to BCC (p < 0.001). In addition, the ratios of PD-L1/CD86 (p < 0.001) and CD28/CD86 (p < 0.001) were significantly higher in cSCC. The invasive front of both tumor entities showed higher expression levels of all immune markers compared to the tumor core in both tumor entities. The significantly higher expression of PD-1, PD-L1, and CD28 in cSCC, along with the predominance of the inhibitory ligand PD-L1 as compared to the activating CD86 in cSCC, provide a potential explanation for the better objective response rates to anti-PD-1 immunotherapy as compared to BCC. Furthermore, the predominant site of interaction between the immune system and the tumor was within the invasive front in both tumor types. [ABSTRACT FROM AUTHOR]

Objective Certain types of bacteria contain lipopolysaccharide (LPS), which can cause widespread inflammation in the body, including the pancreas. Fluvoxamine (FLV), a selective serotonin reuptake inhibitor (SSRI) commonly prescribed for psychiatric disorders, has been shown to possess anti-inflammatory properties and may be beneficial in conditions involving tissue damage and inflammation. This study aims to evaluate the potential protective effects of FLV against experimentally induced pancreatic disease in rats using LPS. Material and Method In this experiment, a total of 32 Wistar albino male rats were randomly divided into four groups: control, LPS (5 mg/kg, intraperitoneally (i.p.)), LPS + FLV (50 mg/kg FLV, i.p.) and FLV. The rats were euthanatized 6 hours after the administration of LPS, and serum and pancreas tissue samples were collected during the necropsy for biochemical, histopathological, and immunohistochemical evaluations. Results According to the study findings, LPS lowered blood glucose levels. Histological examination showed that LPS caused edema, mild infiltration of inflammatory cells, increased vacuolization in the cells of the Langerhans islet, and severe hyperemia. Immunohistochemical investigations revealed a reduction in the expression of insulin and amylin. The biochemical, histopathological, and immunohistochemical outcomes were improved by FLV. Conclusion The results of this experimental rat model study indicated that LPS causes damage to the endocrine pancreas. However, FLV demonstrated significant ameliorative effects on the pancreas in rats with LPS- induced pancreatitis. [ABSTRACT FROM AUTHOR]

CRNDE is an oncogene expressed as a long non-coding RNA. However, our team previously reported that the CRNDE gene also encodes a micropeptide, CRNDEP. The amino acid sequence of CRNDEP has recently been revealed by other researchers, too. This study aimed to investigate genetic alterations within the CRNDEP-coding region of the CRNDE gene, methylation profiling of this gene, and CRNDEP expression analysis. All investigations were performed on clinical material from patients with ovarian tumors of diverse aggressiveness. We found that CRNDEP levels were significantly elevated in highly aggressive tumors compared to benign neoplasms. Consistently, a high level of this micropeptide was a negative, independent, prognostic, and predictive factor in high-grade ovarian cancer (hgOvCa) patients. The cancer-promoting role of CRNDE(P), shown in our recent study, was also supported by genetic and epigenetic results obtained herein, revealing no CRNDEP-disrupting mutations in any clinical sample. Moreover, in borderline ovarian tumors (BOTS), but not in ovarian cancers, the presence of a single nucleotide polymorphism in CRNDE, rs115515594, significantly increased the risk of recurrence. Consistently, in BOTS only, the same genetic variant was highly overrepresented compared to healthy individuals. We also discovered that hypomethylation of CRNDE is associated with increased aggressiveness of ovarian tumors. Accordingly, hypomethylation of this gene's promoter/first exon correlated with hgOvCa resistance to chemotherapy, but only in specimens with accumulation of the TP53 tumor suppressor protein. Taken together, these results contribute to a better understanding of the role of CRNDE(P) in tumorigenesis and potentially may lead to improvements in screening, diagnosis, and treatment of ovarian neoplasms. [ABSTRACT FROM AUTHOR]

Introduction: Oral cancer pretences a substantial global health challenge, with approximately 20 per 100,000 population and accounts for over 30% of all cancers in India. Human papillomavirus (HPV) has emerged as a key risk factor for the development of Head and neck cancer, constituting around 60% of all oral cancer cases around the world. This review aims to present a comprehensive overview of the current relationship between HPV and oral cancer, covering aspects such as detection and prevalence rate. Methods: A comprehensive literature search was conducted using PubMED and Google scholar databases to identify studies published in English from January 2012 to January 2022 that investigated the association between HPV and oral cancer and relevant articles were selected. Results: The literature search yielded 32 studies that met our inclusion criteria. Our analysis revealed that HPV is a common finding in oral cancer, with a pooled prevalence rate of 22.4%. HPV-16, followed by HPV-18 was the prominent genotypes detected in HPV induced Head and neck cancer. Conclusions: HPV considerably contributes to the onset of Head and neck cancer, and identifying it early can serve as an indicator for diagnosis and prognosis which in turn aids to formulate successful prevention and treatment approaches. Immunization against HPV shows potential for decreasing the occurrence of Head and neck cancer linked to HPV. [ABSTRACT FROM AUTHOR]

Accurately distinguishing HER2-2+ tumors from HER2-0/1+ tumors via immunohistochemistry (IHC) is still very challenging. HER2 IHC 2+ is considered to indicate moderate expression and is easier to distinguish, with more reliable results in previous and current clinical practice. We focused on HER2-2+ patients and evaluated the switch in HER2 status between primary and paired recurrent disease patients to evaluate the discordance of HER2-2+ expression. We included patients who were HER2-2+ of primary or rebiopsy tumor samples, to evaluate the evolution of HER2-2+ expression. In the cohort with a total of 159 patients with HER2-2+ expression in either primary tumor or locoregional/distant metastasis samples, 44.0% had HER2-2+ in primary tumor and 88.8% in recurrent disease. Among patients with primary and recurrent HER2-2+ breast cancers, 18.5% and 15.2% of the patients, respectively, had HER2 gene amplification via ISH. The overall rate of discordance in HER2 IHC results was 67.1%. Among primary HER2-2+ patients, 74.6% were maintained in the HER2-2+ cohort at the recurrence. The discordance was mostly driven by patients switching from HER2-2+ to HER2-1+ (64.7%). Among HER2-2+ recurrent patients, discordance in the IHC results was mostly driven by switching from HER2-0 to HER2-2+ (47.1%). When HER2-low was added to the analysis, the overall rate of HER2 discordance was 40.4%. The proportion of patients with discordant HER2 expression was significantly greater among HR-positive patients than negative patients (44.1% vs. 21.7%, p=0.062). HER2 expression in primary and recurrent breast cancer samples was highly unstable. Discordance was more frequently observed in the HR-positive population. [ABSTRACT FROM AUTHOR]

Generating high quality histopathology images like immunohistochemistry (IHC) stained images is essential for precise diagnosis and the advancement of computer-aided diagnostic (CAD) systems. Producing IHC images in laboratory is quite expensive and time consuming. Recently, some attempts have been made based on artificial intelligence techniques (particularly, deep learning) to generate IHC images. Existing IHC stained image generation methods, still have a limited performance due to the complex structures and variations in cells shapes, potential nonspecific staining and variable antibody sensitivity. This paper proposes a novel technique known as harmonic conditional generative adversarial network (HcGAN) for generating high quality IHC-stained images. To generate the IHC images, the HcGAN model is fed with the widely available hematoxylin and eosin (H&E) images that contain cellular and morphological underlying structures of diverse cancer tissues. Such approach helps generate high quality IHC images mimicking the real ones that highlight the positive cells. The proposed HcGAN model is based generative adversarial learning with generator and discriminator networks. In HcGAN, harmonic convolution based on discrete cosine transform filter banks is employed in the generator and discriminator networks instead of the standard convolution in order to improve visual quality of the generated images and address the issue of overfitting. Our qualitative and quantitative results demonstrate that the proposed HcGAN achieved the highest performance over state-of-the-art methods using two publicly available datasets.

Uveal melanoma (UM) is the most common primary intraocular tumor in adults, and despite excellent local control, more than 50% of patients develop and die from metastatic disease. Loss of BAP1 nuclear staining, a surrogate marker of BAP1 mutation, and preferentially expressed antigen in melanoma (PRAME) messenger RNA overexpression, as assessed using qPCR, have previously been shown to correlate with increased metastasis rate in UM. In this study, we demonstrated that UM could be successfully risk-stratified using a combination of BAP1 and PRAME immunohistochemical (IHC) stains. We retrospectively reviewed 318 UM cases with sufficient tissue and performed BAP1 and PRAME IHC to stratify them as BAP1+/PRAME- (group 1, n = 135), BAP1+/PRAME+ (group 2, n = 43), BAP1-/PRAME- (group 3, n = 94), and BAP1-/PRAME+ (group 4, n = 46). Increasing the study risk group on the basis of loss of BAP1 expression and positive PRAME staining was associated with a higher rate of metastasis and disease-specific death and lower metastasis-free survival (MFS) and disease-specific survival (DSS). Among tumors with loss of BAP1 staining, PRAME positivity was associated with shorter MFS (P = .018) and showed a trend toward shorter DSS (P = .061). Among tumors with retained BAP1 staining, PRAME positivity was associated with shorter MFS and DSS (P = .001 and P = .021, respectively). In summary, a combination of BAP1 and PRAME IHC can be used for risk stratification of UMs.

Objective: Alveolar soft part sarcoma (ASPS) is characterized by distinctive histomorphology of variably discohesive epithelioid cells arranged in nests and translocation of t(x;17) (p11.2;q25) resulting in ASPSCR1-TFE3 fusion. The aim of the present study is to review the clinical, histopathological, and immunohistochemical profile of ASPS with a focus on unusual histological features. Material and Method: The present study is retrospective and descriptive. All cases with a diagnosis of ASPS were retrieved with clinical and radiology details. Results: 22 patients of ASPS were identified. The most common site was the lower extremity and the size range was 3-22 cm. 54.5% of the patients had metastasis, with the lung as the most common site. Metastasis preceded detection of primary tumour in two cases. All cases showed similar histopathology of monomorphic epithelioid cells arranged in nests encircled by sinusoidal vasculature. Architecturally, the organoid pattern (81.8%) was followed by the alveolar pattern. 68.2% of the cases showed apple bite nuclei as the predominant nuclear feature. Rare nuclear features included binucleation (n=13), multinucleation (n=8), pleomorphism (n=4), nuclear grooves in three cases and intranuclear inclusion in one case, mitosis (n=5), and focal necrosis (n=6). All cases were positive for TFE3 and negative for AE1/AE3, EMA, HMB45, PAX8, MyoD1, SMA, synaptophysin, and chromogranin. Only two cases showed focal S100 positivity while one showed focal desmin positivity. Conclusion: Diffuse strong nuclear TFE3 positivity is sensitive for ASPS in an appropriate clinicoradiological context. Due to the high propensity for early metastasis, complete metastatic work-up and long term follow up is recommended.

Oral carcinoma cuniculatum (CC) is a rare distinct variant of oral squamous cell carcinoma (OSCC) displaying unique clinical and histopathological features. CC is a locally aggressive malignancy with lower metastatic potential and has a tendency to invade the underlying bone yielding nonspecific clinical presentation resembling fungal/bacterial infections or osteomyelitis. Seven rare cases of gingival CC are reported with special emphasis on immunohistochemistry and special stains. Clinical and demographic profile were analysed and all cases were subjected to Ki-67 and p53 immunostaining and Ayoub-Shklar histochemical staining. For comparison, age and gender matched seven cases of conventional OSCC with bony involvement were recruited. IBM SPSS statistics software version 26 (IBM Analytics, Armonk, New York, U.S.) was used to analyse the data. All cases of CC lacked p53 expression in contrast to OSCC and demonstrated intermediate to high proliferation index in stroma also. The keratin staining and cytokeratin profile in OSCC and CC were also found to be statistically different. CC is a rare well differentiated variant of OSCC with different pathways involved in the pathogenesis. Local aggressive and invasive nature could be the result of unique interaction between epithelium and stromal elements. Oral health care provider must be familiar with this uncommon variant of OSCC. [ABSTRACT FROM AUTHOR]

Prostate cancer (PC), accounts for more than one-fourth of all cancer diagnoses, and the most frequently diagnosed cancer among men in 2022. The immunoglobulin (IG) Program death ligand-1(PD-1) cell surface receptor is predominantly expressed on the surface of many cells. The purpose of this study was to demonstrate the relationship between Program death ligand expression and some aggressive features of prostate cancer including perineural invasion, vascular invasion and necrosis. Thirty cases of prostate cancer with age range from 60 to 80 year old and 30 cases of normal prostate tissue with age under 25 year old were separated into two groups in a retrospective case-control research that encompassed 60 cases. All malignant cases were examined by consultant pathologists for the diagnosis of prostate carcinoma, and each block of tissue was divided into two slides, one for hematoxylin and eosin (H&E) staining and the other for immunoglobulin (IHC) staining of PDL-1. The expression pattern of Program Death Ligand was investigated in these samples and its relationship to particular clinic-pathological characteristics. Despite it was not expressed in healthy prostatic tissue, program death ligand demonstrated to be positive in prostate cancer with vascular invasion, perineural invasion, and necrosis, while it was negative in healthy prostatic tissue. High expression of Program death ligand was correlated with poor differentiation, neural invasion, and vascular invasion; these criteria indicate that the expression of Program death ligand is associated with high grade and aggressive tumors. The current study confirms that perineural invasion, vascular invasion, and necrosis are all accompanied by a rise in Program death ligand expression regardless their grade and stage. [ABSTRACT FROM AUTHOR]

Lung cancer has progressed in the last century from being a rare disease to becoming the most common cancer in men and the second most common cancer in women, as well as the leading cause of cancer related deaths worldwide.1 While it is declining in the developed world, its incidence has increased in developing countries and among women. Newer diagnostic methods, immunohistochemistry and molecular/genetic testing has brought about considerable changes in the classification of lung carcinoma and paved the way for personalized targeted newer therapies. This article aims to highlight the tremendous advances that have occurred in the classification, diagnosis and treatment of bronchogenic carcinoma while addressing its fundamental aspects. [ABSTRACT FROM AUTHOR]

Objective: Alveolar soft part sarcoma (ASPS) is characterized by distinctive histomorphology of variably discohesive epithelioid cells arranged in nests and translocation of t(x;17) (p11.2;q25) resulting in ASPSCR1-TFE3 fusion. The aim of the present study is to review the clinical, histopathological, and immunohistochemical profile of ASPS with a focus on unusual histological features. Material and Methods: The present study is retrospective and descriptive. All cases with a diagnosis of ASPS were retrieved with clinical and radiology details. Results: 22 patients of ASPS were identified. The most common site was the lower extremity and the size range was 3-22 cm. 54.5% of the patients had metastasis, with the lung as the most common site. Metastasis preceded detection of primary tumour in two cases. All cases showed similar histopathology of monomorphic epithelioid cells arranged in nests encircled by sinusoidal vasculature. Architecturally, the organoid pattern (81.8%) was followed by the alveolar pattern. 68.2% of the cases showed apple bite nuclei as the predominant nuclear feature. Rare nuclear features included binucleation (n=13), multinucleation (n=8), pleomorphism (n=4), nuclear grooves in three cases and intranuclear inclusion in one case, mitosis (n=5), and focal necrosis (n=6). All cases were positive for TFE3 and negative for AE1/AE3, EMA, HMB45, PAX8, MyoD1, SMA, synaptophysin, and chromogranin. Only two cases showed focal S100 positivity while one showed focal desmin positivity. Conclusion: Diffuse strong nuclear TFE3 positivity is sensitive for ASPS in an appropriate clinicoradiological context. Due to the high propensity for early metastasis, complete metastatic work-up and long term follow up is recommended. [ABSTRACT FROM AUTHOR]

Neosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between Toxoplasma gondii and Neospora caninum. We used T. gondii and N. caninum recombinant proteins, expressed in Escherichia coli and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti– Neospora -rNcSRS2 and anti– Toxoplasma -rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with T. gondii and N. caninum. The anti– Neospora -rNcSRS2 serum labeled specifically only N. caninum– infected tissue blocks, and the anti– Toxoplasma -rTgSRS2 serum was specific to only T. gondii– infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing. [ABSTRACT FROM AUTHOR]

Mesenchymal neoplasms with GLI1 alterations have recently been reported in several anatomic locations. Their morphology and immunohistochemistry (IHC) are nonspecific, making their recognition a true challenge. To assess the diagnostic value of GLI1 and p16 IHC for identifying GLI1-altered neoplasms, we evaluated 12 such neoplasms (6 GLI1-amplified and 6 with GLI1-fusions) using the GLI1 IHC. Additionally, we evaluated some of their morphological and molecular mimickers, including glomangiomas, Ewing sarcomas (ES), myxoid liposarcomas, and MDM2/CDK4-amplified sarcomas (well-differentiated liposarcoma/WDLPS, dedifferentiated liposarcoma/DDLPS, and intimal sarcoma). All successfully tested GLI1-altered tumors (11/11) demonstrated at least moderate/strong nuclear and/or cytoplasmic GLI1 IHC positivity. GLI1-amplified tumors exhibited a moderate/strong predominantly nuclear staining, compared to a moderate, patchy, and predominantly cytoplasmic GLI1 positivity in GLI1-fusion tumors. Among their mimics, GLI1 immunoreactivity, either cytoplasmic or nuclear, was observed in intimal sarcoma (3/3) and WDLPS/DDLPS (22/25). GLI1 IHC demonstrated 92% sensitivity and 90.8% specificity in diagnosing GLI1-altered neoplasms. Strong/moderate nuclear/cytoplasmic p16 immunoexpression was noted in all GLI1-amplified tumors compared to none of fused cases. Overall, the GLI1/p16 combination demonstrated a sensitivity and specificity of 100% and 93% for GLI1-amplified tumors. In conclusion, we confirm that GLI1 IHC represents a good, quick, and cheap helpful screening tool. The inclusion of p16 may aid in pre-screening for potential GLI1-amplified neoplasms and provide insights on which tumors warrant further molecular testing. [ABSTRACT FROM AUTHOR]

Introduction: Ampullary adenocarcinoma is a rare neoplasm often treated by the complex Whipple's procedure. Several histological factors predict poor prognosis including pancreatobiliary morphology, presence of lymphovascular, perineural invasion and local or distant metastasis. Systemic therapy with gemcitabine, 5-fluorouracil regimens are given with variable benefits. Immunotherapy checkpoint inhibitors have shown beneficial anti-tumor effects in several carcinomas, the most remarkable being in non-small cell lung cancer. Administration of these novel drugs is based on immunohistochemical expression (which may or may not be indicative of response to therapy) along with meticulous decision making by the multidisciplinary team. Immunohistochemistry (IHC) is an effective means of immune marker demonstration and has been used in various tumor types for predictive and prognostic purposes. Methods: PD-L1 IHC (clone E1L3N) was applied in 101 cases of ampullary adenocarcinoma. Tumor infiltrating lymphocytes were also evaluated. The immunoreactivity was assessed and categorized into following staining thresholds: <1%, <5%, <10% and ≥10% for tumor cells (membranous and/or cytoplasmic staining pattern), and 5% and 10% cut-offs for immune cells. Results: We found that at a 10% cut-off, 73.3% (74/101) patients were men (P =.006) older than 50 years of age (P <.001) presenting with a tumor measuring <3 cm (P =.001). It was significantly associated with intestinal differentiation (P =.004) and grade 1 tumors (P =.001). Twelve patients presented with recurrence as well (P =.03). Conclusion: In the context of ampullary adenocarcinoma, this study highlights the positivity observed with the PD-L1 IHC clone E1L3N at different thresholds, with the particularly stronger associations being evident at a 10% cut-off. [ABSTRACT FROM AUTHOR]

Membranous nephropathy (MN) is a common cause of nephrotic syndrome in adults. MN is characterized by subepithelial accumulation of immune complexes along the glomerular basement membrane. The immune complexes are composed of immunoglobulin G and a target antigen. PLA2R is the target antigen in approximately 60% of MN cases, and MN is traditionally classified as PLA2R-positive or PLA2R-negative MN. Over the last 7 years, additional target antigens have been identified, which have specific disease associations, distinctive clinical and pathologic findings, and therapeutic implications. The newly discovered target antigens include NELL1, EXT1/EXT2, NCAM1, SEMA3B, PCDH7, FAT1, CNTN1, NTNG1, PCSK6 and NDNF. To group all these antigens into a generic 'PLA2R-negative' MN group is imprecise and un-informative. We propose a logical approach for detection of the target antigen which includes (i) currently available serology-based testing to detect anti-PLA2R and anti-THSD7A antibodies; and (ii) kidney biopsy testing to detect the target antigens. Determination of the antigen on kidney biopsy can be done by immunohistochemistry or immunofluorescence studies. Alternatively, laser capture microdissection (LCM) of glomeruli followed by mass spectrometry (MS) can be used to identify a target antigen. LCM/MS has the advantage of being a one-stop test and is particularly useful for detection of rare target antigens. At the current time, while it is possible to detect the newer antigens by immunohistochemistry/immunofluorescence/LCM/MS, serology-based tests to detect serum antibodies to the new antigens are not yet available. It is critical that serology-based tests should be developed not just for accurate diagnosis, but as a guide for treatment. We review the current methodology and propose an algorithm for diagnosis and detection of target antigens in MN that may shape the current practice in the future. Membranous nephropathy (MN) results from accumulation of subepithelial immune complexes along the glomerular basement membrane. PLA2R is the most common target antigen, but newly discovered target antigens have filled the void of PLA2R-negative MN. MN associated with the newly discovered target antigens have distinctive clinical and pathologic findings, treatment and prognostic implications. These include NELL1, EXT1/EXT2, NCAM1, PCDH7, SEMA3B, CNTN1, FAT1, NDNF and PCSK6. Immunohistochemistry/immunofluorescence methodology is currently in use for detecting target antigens in kidney biopsy tissue, although we anticipate laser capture microdissection of glomeruli followed by mass spectrometry will become available soon. Serologic testing is currently available for only detecting antibodies to PLA2R and THSD7A. It is critical that serologic tests become available for detecting antibodies to the newly discovered antigens. [ABSTRACT FROM AUTHOR]

Bone metastasis has been reported in up to 70% of patients with advanced breast cancer. A total of 55.76% of skeletal metastases in women were derived from breast cancer. However, patients with bone metastasis from an occult primary breast cancer are a rare subset of patients. Here, we present the case of a 38-year-old woman who had sternum pain for 4 months. A whole-body PET-CT scan revealed that the FDG uptake of both the sternum and internal mammary node was significantly increased. The final diagnosis of occult breast cancer was established by immunohistochemical (IHC) staining, which is of great significance for identifying the origin of a metastatic tumor despite no visualized lesions of mammary glands. [ABSTRACT FROM AUTHOR]

Background and Aim: c-MYC is a potent transcription factor involved in several cellular processes, and is overexpressed in many cancers, including non-Hodgkin lymphoma (NHL). The most common subtype of NHL is diffuse large B-cell lymphoma (DLBCL). DLBCL tumors expressing both the c-MYC and BCL2 proteins, i.e. double expressor (DE), also triple expressors (TE) expressing BCL6 in addition to c-MYC and BCL2, are all more aggressive and refractory to treatment. Our study aims to investigate auxiliary immunohistochemical tests (IHC) not routinely used, namely c-MYC, BCL2 and BCL6, seeking a better prognostic testing protocol for DLBCL patients. Materials and Methods: Forty-one formalin-fixed paraffin-embedded (FFPE) DLBCL samples were collected retrospectively from the pathology laboratory at Al-Assad University Hospital after approval from the ethics committee at Damascus University. Clinical data was obtained from patients records in regards to progression-free survival (PFS) and other parameters. Slides were microtomed from each FFPE sample and used to perform IHC tests of the c-MYC, BCL2 and BCL6 oncoproteins. Fisher’s exact test was used to study the independence of protein IHC results in relation to one another and compared with PFS. Results: Statistical analysis of the c-MYC overexpression and double expressor status showed no correlation (P>0.05) with PFS in DLBCL patients. Whereas, when BCL6 IHC results were analyzed, BCL6 positivity and triple expressor status showed a significant association (P

Background: Timely reporting of molecular biomarkers is critical in guiding optimal treatment decisions in patients with advanced non-small cell lung carcinoma (NSCLC). Any delays along the tissue or treatment pathway may be associated with suboptimal treatment/outcomes and a reduced quality of life. For many centres, biomarkers are tested off-site. Methods: A retrospective chart review of 123 patients with advanced NSCLC seen between 1 June 2021 and 30 June 2022 was conducted. With a focus on core biomarkers (PD L1, EGFR, and ALK), the outcome variables were as follows: total turnaround time (total TAT), divided into pre-laboratory, laboratory, and post-laboratory time intervals, as well as time to treatment decision (TOTD) and time to optimal systemic therapy decision (TOTSD). Results: At first consult, only 20.3% of patients had all core biomarker results available. The median total TAT was significantly longer for non-squamous (non-SCC) than squamous cell carcinoma (SCC) specimens (36.5 versus 22 days, p < 0.001). The median pre-laboratory time for the entire cohort was 5 calendar days. The median laboratory testing time was greater for non-SCC compared to the SCC specimens (23 versus 12 days, p < 0.001). The median time from consult to TOTD was 19 calendar days for the entire cohort. Conclusions: This study emphasizes the need for the expansion of regional resources to meet the clinical needs of advanced NSCLC patients treated at a regional cancer centre which uses an off-site molecular laboratory.

Background & aimsThe stimulator of interferon genes (STING)/TANK-binding kinase 1 (TBK1) pathway is vital in mediating innate immune and inflammatory responses during oxidative/endoplasmic reticulum (ER) stress. However, it remains unknown whether macrophage thioredoxin-interacting protein (TXNIP) may regulate TBK1 function and cell death pathways during oxidative/ER stress.MethodsA mouse model of hepatic ischaemia/reperfusion injury (IRI), the primary hepatocytes, and bone marrow-derived macrophages were used in the myeloid-specific TXNIP knockout (TXNIPM-KO) and TXNIP-proficient (TXNIPFL/FL) mice.ResultsThe TXNIPM-KO mice were resistant to ischaemia/reperfusion (IR) stress-induced liver damage with reduced serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) levels, macrophage/neutrophil infiltration, and pro-inflammatory mediators compared with the TXNIPFL/FL controls. IR stress increased TXNIP, p-STING, and p-TBK1 expression in ischaemic livers. However, TXNIPM-KO inhibited STING, TBK1, interferon regulatory factor 3 (IRF3), and NF-κB activation with interferon-β (IFN-β) expression. Interestingly, TXNIPM-KO augmented nuclear factor (erythroid-derived 2)-like 2 (NRF2) activity, increased antioxidant gene expression, and reduced macrophage reactive oxygen species (ROS) production and hepatic apoptosis/necroptosis in IR-stressed livers. Mechanistically, macrophage TXNIP deficiency promoted cylindromatosis (CYLD), which colocalised and interacted with NADPH oxidase 4 (NOX4) to enhance NRF2 activity by deubiquitinating NOX4. Disruption of macrophage NRF2 or its target gene 2',5' oligoadenylate synthetase-like 1 (OASL1) enhanced Ras GTPase-activating protein-binding protein 1 (G3BP1) and TBK1-mediated inflammatory response. Notably, macrophage OASL1 deficiency induced hepatocyte apoptotic peptidase activating factor 1 (APAF1), cytochrome c, and caspase-9 activation, leading to increased caspase-3-initiated apoptosis and receptor-interacting serine/threonine-protein kinase 3 (RIPK3)-mediated necroptosis.ConclusionsMacrophage TXNIP deficiency enhances CYLD activity and activates the NRF2-OASL1 signalling, controlling IR stress-induced liver injury. The target gene OASL1 regulated by NRF2 is crucial for modulating STING-mediated TBK1 activation and Apaf1/cytochrome c/caspase-9-triggered apoptotic/necroptotic cell death pathway. Our findings underscore a novel role of macrophage TXNIP-mediated CYLD-NRF2-OASL1 axis in stress-induced liver inflammation and cell death, implying the potential therapeutic targets in liver inflammatory diseases.Lay summaryLiver inflammation and injury induced by ischaemia and reperfusion (the absence of blood flow to the liver tissue followed by the resupply of blood) is a significant cause of hepatic dysfunction and failure following liver transplantation, resection, and haemorrhagic shock. Herein, we uncover an underlying mechanism that contributes to liver inflammation and cell death in this setting and could be a therapeutic target in stress-induced liver inflammatory injury.

Background: Timely reporting of molecular biomarkers is critical in guiding optimal treatment decisions in patients with advanced non-small cell lung carcinoma (NSCLC). Any delays along the tissue or treatment pathway may be associated with suboptimal treatment/outcomes and a reduced quality of life. For many centres, biomarkers are tested off-site. Methods: A retrospective chart review of 123 patients with advanced NSCLC seen between 1 June 2021 and 30 June 2022 was conducted. With a focus on core biomarkers (PD L1, EGFR, and ALK), the outcome variables were as follows: total turnaround time (total TAT), divided into pre-laboratory, laboratory, and post-laboratory time intervals, as well as time to treatment decision (TOTD) and time to optimal systemic therapy decision (TOTSD). Results: At first consult, only 20.3% of patients had all core biomarker results available. The median total TAT was significantly longer for non-squamous (non-SCC) than squamous cell carcinoma (SCC) specimens (36.5 versus 22 days, p < 0.001). The median pre-laboratory time for the entire cohort was 5 calendar days. The median laboratory testing time was greater for non-SCC compared to the SCC specimens (23 versus 12 days, p < 0.001). The median time from consult to TOTD was 19 calendar days for the entire cohort. Conclusions: This study emphasizes the need for the expansion of regional resources to meet the clinical needs of advanced NSCLC patients treated at a regional cancer centre which uses an off-site molecular laboratory. [ABSTRACT FROM AUTHOR]

Background & Objective: Colorectal carcinoma (CRC) is one of the most common cancers worldwide. The interaction of programmed cell death receptor 1 (PD-1) and programmed death ligand 1 (PD-L1) plays an important role by inhibiting the immune mechanism by which cancer cells escape antitumor immunity. Immunotherapy using checkpoint inhibitors is a growing treatment modality in many cancers; one such is anti-PD1/PD-L1. The present study aimed to study the immunohistochemical (IHC) expression of PD-L1 in CRC and its association with various known clinicopathological parameters. Methods: This study was a 2-year prospective study and included 34 colectomy specimens diagnosed as colorectal adenocarcinoma. The expression of PD-L1 was evaluated on tumoral cells and tumor-infiltrating immune cells (TIICs) and was correlated with various clinicopathological parameters. Results: Immunohistochemical expression of PD-L1 on tumoral cells and tumor microenvironment in CRC revealed positivity in 17.65% of cases each. The PD-L1 expression on tumoral cells was associated with lymphovascular invasion (LVI) and perineural invasion (PNI) with P-values of 0.012 and 0.005, respectively, while PD-L1 expression on TIICs was associated with tumor budding with a P-value of 0.022. Conclusion: IHC expression of PD-L1 on tumoral cells and immune cells may be associated with some known poor prognostic factors. Since anti-PD1/PD-L1 is used for targeted therapy, it may be beneficial and economically feasible to evaluate PD-L1 in CRC and establish its role as a prognostic factor. [ABSTRACT FROM AUTHOR]

Breast cancer (BC), characterized by its diverse molecular profiles and clinical outcomes, presents a significant challenge in the development of effective therapeutic strategies. Metabolic reprogramming, a defining characteristic of cancer, has emerged as a promising target for novel therapies. SLC7A11, an amino acid transporter that facilitates cysteine uptake in exchange for glutamate, plays a crucial role in sustaining the altered metabolism of cancer cells. This study delves into the comprehensive analysis of SLC7A11 at the genomic, transcriptomic, and protein levels in extensive BC datasets to elucidate its potential role in different BC subtypes. SLC7A11 gene copy number and mRNA expression were evaluated using the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) cohort (n = 1,980) and Breast Cancer Gene Expression Miner (n = 4,712). SLC7A11 protein was assessed using immunohistochemistry in a large BC cohort (n = 1,981). Additionally, The Cancer Genome Atlas (TCGA) dataset was used to explore SLC7A11 DNA methylation patterns using MethSurv (n = 782) and association of SLC7A11 mRNA expression with immune infiltrates using TIMER (n = 1,100). High SLC7A11 mRNA and SLC7A11 protein expression were significantly associated with high tumor grade (p = .02), indicating a potential role in cancer progression. Interestingly, SLC7A11 copy number gain was observed in HER2+ tumors (p = .01), suggesting a subtype-specific association. In contrast, SLC7A11 mRNA expression was higher in the basal-like/triple-negative (TN; p < .001) and luminal B tumors (p = .02), highlighting its differential expression across BC subtypes. Notably, high SLC7A11 protein expression was predominantly observed in Estrogen Receptor (ER)-negative and Triple Negative (TN) BC, suggesting a role in these aggressive subtypes. Further analysis revealed that SLC7A11 was positively correlated with other amino acid transporters and enzymes associated with glutamine metabolism, implying a coordinated role in metabolic regulation. Additionally, SLC7A11 gene expression was positively associated with neutrophil and macrophage infiltration, suggesting a potential link between SLC7A11 and tumor immunity. Our findings suggest that SLC7A11 plays a significant role in BC metabolism, demonstrating differential expression across subtypes and associations with poor patient outcomes. Further functional studies are warranted to elucidate the precise mechanisms by which SLC7A11 contributes to BC progression and to explore its potential as a therapeutic target. [ABSTRACT FROM AUTHOR]

Background: Immunohistochemistry (IHC) plays an important role in the management and outcome of breast cancer. It has been observed that the responses to same chemotherapy or hormone therapy are different among cases of breast cancer. Despite the recognition of the IHC markers and appropriate treatment, poor response or relapse can be attributed to the discordance between marker status of breast tumors and axillary lymph nodes. Therefore, our aim is to assess the molecular markers in primary breast cancer/tumor (PBT) and metastatic axillary lymph nodes (ALN) and their correlation with outcome in patients undergoing treatment for carcinoma breast in our hospital. Methods: This Prospective Observational study was done in an institution in northern India dedicated for people of low socioeconomic strata. Study was done over a period of two years. 85 patients with operable breast carcinoma without neoadjuvant chemotherapy were included in the study. Results: Mean value of age (years) of patients was 50.73 ± 9.8 Majority (96.47%) of patients had invasive breast carcinoma -- no special type. 77.65% of patients had stage IIA breast carcinoma at the time of presentation. IHC of primary tumor correlated with that of axillary lymph nodes mostly and discordance was statistically insignificant. Conclusion: We concluded that discordance in immunohistochemistry profile of primary breast tumor and axillary lymph nodes are statistically insignificant and it is not the reason of poor outcome in certain cases of carcinoma breast. [ABSTRACT FROM AUTHOR]

The immune microenvironment plays an important role in the regulation of diseases. The characterization of the cellular composition of immune cell infiltrates in diseases and respective models is a major task in pathogenesis research and diagnostics. For the assessment of immune cell populations in tissues, fluorescence-activated cell sorting (FACS) or immunohistochemistry (IHC) are the two most common techniques presently applied, but they are cost intensive, laborious, and sometimes limited by the availability of suitable antibodies. Complementary rapid qPCR-based approaches exist for the human situation but are lacking for experimental mouse models. Accordingly, we developed a robust, rapid RT-qPCR-based approach to determine and quantify the abundance of prominent immune cell populations such as T cells, helper T (Th) cells, cytotoxic T cells, Th1 cells, B cells, and macrophages in mouse tissues. The results were independently validated by the gold standards IHC and FACS in corresponding tissues and showed high concordance. [ABSTRACT FROM AUTHOR]

Deletion of CDKN2A occurs in 50% of glioblastomas (GBM), and IFNA locus deletion in 25%. These genes reside closely on chromosome 9. We investigated whether CDKN2A and IFNA were co-deleted within the same heterogeneous tumour and their prognostic implications. We assessed CDKN2A and IFNA14 deletions in 45 glioma samples using an in-house three-colour FISH probe. We examined the correlation between p16INK4a protein expression (via IHC) and CDKN2A deletion along with the impact of these genomic events on patient survival. FISH analyses demonstrated that grades II and III had either wildtype (wt) or amplified CDKN2A/IFNA14, whilst 44% of GBMs harboured homozygous deletions of both genes. Cores with CDKN2A homozygous deletion (n = 11) were negative for p16INK4a. Twenty p16INK4a positive samples lacked CDKN2A deletion with some of cells showing negative p16INK4a. There was heterogeneity in IFNA14/CDKN2A ploidy within each GBM. Survival analyses of primary GBMs suggested a positive association between increased p16INK4a and longer survival; this persisted when considering CDKN2A/IFNA14 status. Furthermore, wt (intact) CDKN2A/IFNA14 were found to be associated with longer survival in recurrent GBMs. Our data suggest that co-deletion of CDKN2A/IFNA14 in GBM negatively correlates with survival and CDKN2A-wt status correlated with longer survival, and with second surgery, itself a marker for improved patient outcomes. [ABSTRACT FROM AUTHOR]

Background: Oral squamous cell carcinoma (OSCC) entails quite noteworthy morbidity and mortality rates instead of immense amount of research and advances. Mutation in the TP53 gene is the most common genetic change found in oral squamous cell carcinoma (OSCC). Once activated, the p53 protein can induce the growth arrest, as well as cell death. Mutation and subsequent inactivation of a tumor suppressor gene causes ‘loss of function’. This can damage its deoxyribonucleic acid (DNA) binding properties and transcription factor function, thus, inhibiting its normal function in cell cycle control and in cell proliferation. Aim: A comparative study of P53 expression in oral squamous cell carcinoma using immunohistochemical techniques. Materials and Methods: To study the relationship between histological grades of oral squamous cell carcinoma and P53 protein expression and also to compare along with normal mucosa, a total of 40 cases of histologically diagnosed OSCC were considered for the study group and 10 paraffin embedded normal mucosal tissue blocks were collected. The analysis of p53 positive cells was performed on IHC stained sections. Only staining of nucleus of epithelial cells was observed. Nuclei with clear brown colour regardless of staining intensity were considered as p53 positive. Counting of cells was done with graticule eyepiece. All these cases were reviewed and classified according to modified Anneroths histological malignancy grading system. Results: When p53 expression was compared between the two groups i.e control and study groups. The expression of p53 was nil in normal oral mucosal tissue (control), while study group i.e OSCC expressed about 47.49% with Std.Dev of ±35.83%. And it was found to be a statistically significant (p<0.001). Conclusion: The present study concludes a positive correlation between the histopathological grades of OSCC and immunohistochemistry (p53 expression). Expression of p53 above the basal layer is an early event in oral carcinogenesis and an indicator of a developing carcinoma preceding morphological tissue alterations. However, since immunohistochemistry cannot always detect changes in p53 expression in premalignant/malignant lesions, it is strongly recommended that p53 immunohistochemistry be used in conjugation with routine histology to increase the sensitivity of detection of cases that eventually progress to malignancy. [ABSTRACT FROM AUTHOR]

Background: Approximately 20% of breast cancers express HER2-positive receptors in the USA. HER2 receptor immunohistochemistry (IHC) staining with equivocal (2+) results commonly undergoes fluorescence in-situ hybridization (FISH) for further classification. Current guidelines do not recommend routine FISH testing in IHC-negative (0 or 1+) cases. This study investigates an institution that performs both IHC and FISH testing on all cases to identify the true HER2-positive rate. Patients and Methods: A retrospective chart review from 2015 to 2021 was conducted at an institution where both HER2 IHC and FISH testing were performed at the time of diagnosis for all invasive breast cancers. The rate of true HER2-positive patients was determined, and patient and tumor characteristics were further explored. Results: A total of 1835 invasive breast cancer cases were primarily treated at this institution. A total of 289 cases were HER2 positive on IHC and FISH testing (15.7%). An additional 38 cases were identified as HER2 negative on IHC, but reclassified as HER2 positive on reflex FISH testing. Total HER2 positive cases increased from 289 (15.7%) to 327 cases (17.8%) with reflex FISH testing. Conclusions: The additional HER2-positive cases after completing FISH testing on IHC-negative tumors suggests there may be a role for routine FISH testing in addition to standard IHC staining to determine HER2 status for breast cancer. The ethical, prognostic and even benefits of a correct diagnosis outweigh the added expense of FISH testing. [ABSTRACT FROM AUTHOR]

Breast cancer is a prevalent disease worldwide and the accurate diagnosis and prognosis of breast cancer are essential for the development of effective treatment plans. Pathology remains the gold standard for diagnosis and prognosis but with limitations such as time-consuming manual scoring and some error-prone results. Recently, deep learning techniques, especially convolutional neural networks (CNN), have been proposed for the interpretation of immunohistochemistry (IHC) results in breast cancer. The objective of this systematic review is to critically assess the existing literature on computer-aided systems for the interpretation of IHC results in breast cancer based on deep learning algorithms. We included studies with models that use novel approaches such as deep learning for quantitative measurements of immunohistochemically stained Ki-67, ER, PR, and HER2 images. We systematically searched PubMed, Scopus, and web of science up to September 2022. 15 studies (seven HER2, seven Ki67, and one ER/PR scoring studies) met our inclusion criteria. Various AI-based assays have been developed for different applications in breast pathology, including diagnostic and prognostic applications, as well as predictive values and responses to treatment. These algorithms have shown promise in improving the accuracy of breast cancer diagnosis and prognosis. It is essential to consider the differences in training and inter-observer variability while designing tools, and there is an urgent need to integrate the detection and analysis of various biomarkers at the same place and time to facilitate the formation of patients' reports and treatment.

Background Papillary thyroid carcinoma is the most common endocrine and thyroid malignancy. The (BRAF V600E) mutation is the most common mutation in papillary thyroid cancer (PTC). The mutation is detected in about (60%) of the cases. Patients with BRAF mutation have been linked to aggressive clinical behavior, high recurrence rate, metastasis, and failure of treatment Aim of the study This study aims to find whether Immunohistochemisrty can substitute molecular analysis in detection of BRAF V600E mutation in papillary thyroid carcinoma. This will safely time, cost, and effort. Patients and method This was a retrospective study, carried out in Basrah province, wax blocks were collected from histopathology labs during period from October through September 2023. Tissue sections were obtained from the paraffin blocks and submitted to routine hematoxylin and eosin stain. Then Immunohistochemical staining using primary monoclonal antibody for detection of BRAF mutation in papillary thyroid carcinoma. After examination of H &E slides, the area of papillary thyroid carcinoma was determined and Then piece of tumor tissue with thickness of 5~10 um and a surface Darweesh, S., Al-Diab, J., Al-Harron, S., Hasson, H. Comparative study between Immunohistochemistry and Molecular method in detection of BRAF V600E mutation in papillary thyroid carcinoma. Basrah Journal of Surgery, 2023; 29(2): 76-87. doi: 10.33762/bsurg.2023.144308.1062 77 area between 0.5~1 cm². was taken from the wax block for DNA extraction and detection of BRAF mutation using Real-time PCR. Results Thirty cases of papillary thyroid carcinoma were included in this study. Twenty-five case (83%) were female and five cases (16.66%) were male with mean age of 41.33+16.1 (range 16-81 years). Using real-time PCR, twelve cases (40%) were positive for BRAF mutation and eighteen cases (60%) were negative for BRAF mutation. Using immunohistochemistry, thirteen cases (43.3%) were positive for BARF mutation & seventeen cases (56.7%) were negative for the mutation. The sensitivity of Immunohistochemistry in detecting BRAF mutation in papillary thyroid carcinoma is (66.7%) with specificity (72.2%). The positive predictive value is (61.5%) and the negative predictive value is (76.5%). Conclusion The findings from this study reveal a discrepancy between Immunohistochemistry and real-time PCR in identifying the BRAF V600E mutation. It is important to note that Immunohistochemistry should not serve as a substitute for molecular methods in detecting this mutation. In fact, relying solely on Immunohistochemistry may not be reliable due to its lower sensitivity. This could potentially lead to increased time, cost, and effort in the diagnostic process

Introduction: Gastrointestinal stromal tumors (GIST) are the most common mesenchymal neoplasms of the gastrointestinal tract and their diagnosis is mainly based on histopathology and immunohistochemical study with CD117 marker. However, there are many GIST tumors that are CD117 negative and for such cases come the role of other IHC markers. The study aimed to observe the various histological features and demographic profiles of GISTs and the role of various immunohistochemical markers in the confirmation of diagnosis. Methodology: A hospital-based observational study was conducted on 40 tumor resection materials diagnosed as GIST from April 2017 to February 2023 at tertiary care center K.E.M. Hospital, Pune. All the cases underwent histopathological examination with standard procedure of tissue processing, staining and IHC with CD117. Those cases who were morphologically designated as GIST on HPE were evaluated with a panel of IHC markers like C-KIT, CD34, SMA, desmin, S-100 and vimentin. Results: 35 out of 40 cases (87.5%) were found to be positive for CD117 while IHC markers like CD34, SMA, desmin, S-100 and vimentin were found to be positive in 60%, 20%, 5%, 2.5% and 80% cases respectively. Also, we found significant relationship between histopathological groups different progressive disease risk groups with necrosis, cytologic atypia, cellularity and mucosal invasion (p-value

The establishment of immunotherapy applying immune checkpoint inhibitors (ICI) has provided an important new option for the treatment of solid malignant diseases. However, different tumor entities show dramatically different responses to this therapy. BCC responds worse to anti-PD-1 ICIs as compared to cSCC. Differential immune checkpoint expression could explain this discrepancy and, therefore, the aim of this study was to analyze activating and inhibitory immune checkpoints in cSCC and BCC tissues. Tissue microarrays of the invasive front as well as the tumor core of BCC and cSCC samples were used to evaluate PD-1, PD-L1, CD28, and CD86 expression and their topographic distribution profiles by chromogenic immunohistochemistry. QuPath was used to determine the labeling index. The expression of PD-1, PD-L1, and CD28 was significantly higher in both the tumor core and the invasive front of cSCC samples as compared to BCC (p < 0.001). In addition, the ratios of PD-L1/CD86 (p < 0.001) and CD28/CD86 (p < 0.001) were significantly higher in cSCC. The invasive front of both tumor entities showed higher expression levels of all immune markers compared to the tumor core in both tumor entities. The significantly higher expression of PD-1, PD-L1, and CD28 in cSCC, along with the predominance of the inhibitory ligand PD-L1 as compared to the activating CD86 in cSCC, provide a potential explanation for the better objective response rates to anti-PD-1 immunotherapy as compared to BCC. Furthermore, the predominant site of interaction between the immune system and the tumor was within the invasive front in both tumor types.

Abstract Background The role of human parvovirus B19 (B19V) infection in malignant and benign lesions such as head and neck squamous cell carcinomas (HNSCCs) and oral mucocele lesions has not been established. Herein, we examined, for the first time, the presence of B19V in HNSCCs from Iranian subjects. Methods One hundred and eight HNSCC specimens were analyzed for the presence of B19V using nested polymerase chain reaction (nPCR) and TaqMan quantitative PCR assays. Immunohistochemistry procedures were performed to evaluate the expression of B19V VP1/VP2 proteins, p16INK4a, and NF-κB in tumor tissues and their adjacent non-tumor tissues. In addition, 40 oral mucocele, 30 oral buccal mucosa swabs, and 30 nasopharyngeal swabs obtained from healthy adults were analyzed as controls. Results B19V DNA was detected in 36.1% of HNSCCs. Further, 23.3% of HNSCC specimens showed immunoreactivity against B19V VP1/VP2 proteins. There was a significant difference in the frequency of B19V DNA-positive cases between the patient and control groups (p

Abstract Triple‐negative breast cancer (TNBC) is usually an aggressive disease with a poor prognosis and limited treatment options. The neurotrophic tyrosine receptor kinase (NTRK) gene fusions are cancer type‐agnostic emerging biomarkers approved by the Food and Drug Administration (FDA), USA, for the selection of patients for targeted therapy. The main aim of our study was to investigate the frequency of NTRK aberrations, i.e. fusions, gene copy number gain, and amplification, in a series of TNBC using different methods. A total of 83 TNBCs were analyzed using pan‐TRK immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), real‐time polymerase chain reaction (RT‐PCR), and RNA‐based next‐generation sequencing (NGS). Of 83 cases, 16 showed pan‐TRK positivity although no cases had NTRK‐fusions. Indeed, FISH showed four cases carrying an atypical NTRK1 pattern consisting of one fusion signal and one/more single green signals, but all cases were negative for fusion by NGS and RT‐PCR testing. In addition, FISH analysis showed six cases with NTRK1 amplification, one case with NTRK2 copy number gain, and five cases with NTRK3 copy number gain, all negative for pan‐TRK IHC. Our data demonstrate that IHC has a high false‐positive rate for the detection of fusions and molecular testing is mandatory; there is no need to perform additional molecular tests in cases negativity for NTRK by IHC. In conclusion, the NTRK genes are not involved in fusions in TNBC, but both copy number gain and amplification are frequent events, suggesting a possible predictive role for other NTRK aberrations.

Lu Han,1,2,&ast; Yaping Zhang,2– 4,&ast; Li Li,2,&ast; Qun Zhang,2 Zhihao Liu,2,3 Haiqing Niu,1,2 Jing Hu,2 Zhou Ding,2 Xiao Shi,2 Xiaoping Qian1,2 1Department of Oncology, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China; 2The Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University, Nanjing, People’s Republic of China; 3Department of Oncology, Nanjing Drum Tower Hospital Clinical College of Xuzhou Medical University, Nanjing, People’s Republic of China; 4Department of Pathology, The First People’s Hospital of Yangzhou, Yangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiaoping Qian, Department of Oncology, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, 321 Zhongshan Road, Nanjing, Jiangsu, 210008, People’s Republic of China, Tel +86-13951743162, Fax +86-25-68182342, Email xiaopingqian@nju.edu.cnPurpose: Exploring the expression and prognosis of mismatch repair proteins and PD-L1 in colorectal cancer.Patients and Methods: A total of 272 patients with surgically resected CRC were enrolled in the study from January 2018 to May 2022 at Nanjing Drum Tower Hospital (The Affiliated Hospital of Nanjing University Medical School). Surgically resected samples were collected from patients along with general, clinicopathological, and imaging data for each patient. Immunohistochemistry (IHC) was used to detect expression of MSH2, MSH6, MLH1, and PMS2 proteins in tumor tissue. X-squared (X2) testing was performed to investigate the correlation between expression of MMR proteins and PD-L1 in CRC tumor tissues and clinicopathological characteristics. Correlation analysis was also used to compare the deletion of four MMR proteins in CRC tumor tissues. A survival curve and Log rank test were used to investigate the relationship between the expression of MMR proteins and PD-L1 with regard to CRC patient prognosis and survival.Results: MMR protein expression deletion was correlated with tumor location, the degree of tissue differentiation, and TNM stage (P< 0.05). PD-L1 expression was correlated with TNM stage (P

The HER2 gene is a biomarker for breast cancer prognosis and treatment. Overexpression of HER2 protein determined by immunohistochemistry (IHC) or amplification of the HER2 gene determined by fluorescence in situ hybridization (FISH) is a condition for qualifying patients for anti-HER2 therapy. Due to the high toxicity of anti-HER2 treatment, proper patient selection is essential. In our study we compared 40 cases with IHC staining of HER2 antibody determined by Ventana PATHWAY anti-HER2/neu antibody (4B5) as HER2 2+ with the new antibody (HercepTest™ mAb PharmDx [Dako Omnis] [GE001]). Then using a double-blind study we compared the (IHC) evaluation with FISH results. In 65% of cases (26/40) the IHC 2+ score remained unchanged, in 32.5% of cases (13/40) expression of HER2 protein after IHC with new antibody was indicated as 3+ score, and in one case we observed a decrease of HER2 protein expression to 1+. In all cases but one, in which we found IHC HER 3+ with new antibody, there was FISH amplification. We have reason to believe that the new antibody will reduce the diagnostic time and avoid unnecessary costs. Due to the small study group, further investigation is needed.

In Brazil, the Brazilian Symposium on Human Factors in Computing Systems (IHC) gathers the scientific community of researchers interested in the field of Human-Computer Interaction since 1998, being the main Brazilian event in this sub-area of Computing. Over twenty-one editions, the IHC received works from researchers from different regions of the country who, over the years, have been building their own co-authorship relationships with the other authors of the Symposium. In this context, this paper analysed the IHC from the perspective of those who helped to consolidate this important national scientific event, as well as in the expansion of the Human-Computer Interaction area in the Brazilian scenario, that is, its researchers-authors. In total, 1,443 authors were identified and analysed in the study presented in this work, which considered 873 publications of three IHC tracks: Full Papers, Short Papers, and Innovative Ideas and Emerging Results. Issues related to the publications and to the co-authorship relationships of these authors over the years and in the different article tracks of the IHC were considered. In order to describe their research trajectories within the IHC itself, the study presents, in different scales of time, how these authors evolved in relation to their contributions over time. In addition, this paper analyses how the authors contributed with each other and originated the complex collaboration network of the IHC. For this, co-authorship networks and groups of authors who published together were explored, aiming to clarify the collaborations between these authors, as well as how they evolved until the edition of 2022. In this sense, this work seeks, with each research question, to simplify the presentation of results through different visualizations, which were planned and created to describe information that are not clearly evident when observing the IHC publications in a “disconnected” manner. The results of this study are revealed, described and analysed under different perspectives, as well as discussed in details in this paper.

In Egypt, ostrich farming has had a great increase during the last few years. The most common problems facing ostrich farmers are a high percentage of late embryonic mortality and low hatchability. This study was designed to survey the main causes of late in-shell ostrich embryonic death. A total of 100 freshly dead inshell ostrich embryos, all of them were in the second half of the incubation period with different ages ranging from 25 to 39 days old, were received during the breeding seasons of the period from October 2018- October 2021, from different ostrich farms in Egypt, suffered from late-stage embryonic death during artificial incubation, respiratory signs and diarrhea in breeders, moreover, the surveyed ostrich farms were near to chicken farms suffered from Newcastle Disease outbreaks. Gross findings in all embryos were recorded as anasarca, severe general congestion, and hemorrhages. In this study, 66 dead shell ostrich embryos out of 100 (66%) were immunohistochemically positive for Newcastle Disease Virus. Four Velogenic Newcastle Disease Virus strains (genotype VIIb) were molecularly characterized in 4 immune-positive ND embryos (from 4 different ostrich farms in 4 different Egyptian governorates) by RT-PCR and F gene sequencing. In addition, all embryos were negative for bacterial isolation except for one vNDV positive embryo (1%) (25 days old) that was superinfected by Citrobacter spp. The main histopathological lesions of vNDV-positive ostrich embryos were general edema, extensive congested blood vessels, hemorrhages, necrosis, syncytial cells formation in the upper respiratory tract, and leukocytic cells infiltrations. To the best of our knowledge, this study is the first report of the Velogenic Newcastle Disease Virus that immunohistochemically and molecularly characterized in dead ostrich embryos in Egypt, therefore, Newcastle Disease Virus vaccination of ostrich breeders is strongly recommended. [ABSTRACT FROM AUTHOR]

Background: Lymphocyte-activation gene 3 (LAG3) is an immune checkpoint receptor; novel LAG3 immune checkpoint inhibitors (ICIs) exhibit therapeutic activity in melanoma. The role of LAG3and ICIs of LAG3 are unknown in malignant pleural mesothelioma (MPM). This study aimed to uncover the prognostic landscape of LAG3 in multiple cancers and investigate the potential of using LAG3 as an ICIs target in patients with MPM. Methods: We used The Cancer Genome Atlas (TCGA) cohort for assessing mRNA expression and our cohort for immunohistochemical expression. TCGA cohort were analyzed using the Wilcoxon rank-sum test to compare mRNA expression between normal and tumor tissues in multiple cancers. We used 86 MPM cases from TCGA and 38 MPM cases from our cohort to analyze the expression of LAG3 in tumor-infiltrating lymphocytes. The mean LAG3 mRNA expression was set as the cut-off and samples were classified as positive/negative for immunohistochemical expression. Overall survival (OS) of patients with MPM was determined using the Kaplan–Meier method based on LAG3 mRNA and immunohistochemical expression. OS analysis was performed using the multivariate Cox proportional hazards model. The correlation of LAG3 expression and mRNA expression of tumor immune infiltration cells (TIICs) gene markers were estimated using Spearman correlation. To identify factors affecting the correlation of LAG3 mRNA expression, a multivariate linear regression model was performed. Results: LAG3 mRNA was associated with prognosis in multiple cancers. Elevated LAG3 mRNA expression was correlated with a better prognosis in MPM. LAG3 expression was detected immunohistochemically in the membrane of infiltrating lymphocytes in MPM. LAG3 immunohistochemical expression was correlated with a better prognosis in MPM. The multivariate Cox proportional hazards model revealed that elevated LAG3 immunohistochemical expression indicated a better prognosis. In addition, LAG3 mRNA expression was correlated with the expression of various gene markers of TIICs, the most relevant to programmed cell death 1 (PD-1) with the multivariate linear regression model in MPM. Conclusions: LAG3 expression was correlated with prognosis in multiple cancers, particularly MPM; LAG3 is an independent prognostic biomarker of MPM. LAG3 regulates cancer immunity and is a potential target for ICIs therapy. PD-1 and LAG3 inhibitors may contribute to a better prognosis in MPM. Trial registration: This study was registered with UMIN000049240 (registration day: August 19, 2022) and approved by the Institutional Review Board (approval date: August 22, 2022; approval number: 2022–0048) at Tokyo Women's Medical University. [ABSTRACT FROM AUTHOR]

A neurofibroma is a benign peripheral nerve sheath tumor. They occur in combination with neurofibromatosis or as a solitary mass. Intraoral neurofibromas pose diagnostic difficulties, suggesting the appropriate use of diagnostic markers and clinical knowledge. Here, we report a case of intraoral neurofibroma in a 57-year-old female who reported with the complaint of painless growth in the lower left back tooth region for the past three years. Based on the clinical features, provisional diagnosis of traumatic fibroma was made. However, histopathology proved it to be a benign spindle cell lesion; upon further investigation by immunohistochemistry, it was diagnosed to be a case of neurofibroma. [ABSTRACT FROM AUTHOR]

Kidney renal clear cell carcinoma (KIRC) is the most common type of kidney cancer and its pathogenesis is strongly associated with VHL–HIF–VEGF signaling. SHH ligand is the upstream SHH pathway regulator, while GLI1 is its major effector that stimulates as a transcription factor, i.a. expression of VEGFA gene. The aim of present study was to assess the prognostic significance of SHH, GLI1 and VEGFA immunoreactivity in KIRC tissues. The analysis included paired tumor and normal samples from 34 patients with KIRC. The immunoreactivity of SHH, GLI1 and VEGFA proteins was determined by immunohistochemical (IHC) renal tissues staining. The IHC staining results were assessed using the immunoreactive score (IRS) method which takes into account the number of cells showing a positive reaction and the intensity of the reaction. Increased GLI1 protein immunoreactivity was observed in KIRC tissues, especially in early-stage tumors, according to the TNM classification. Elevated expression of the VEGFA protein was noted primarily in high-grade KIRC samples according to the Fuhrman/WHO/ISUP scale. Moreover, a directly proportional correlation was observed between SHH and VEGFA immunoreactivity in TNM 3 + 4 and Fuhrman/ISUP/WHO 3 + 4 tumor tissues as well as in samples of patients with shorter survival. We also observed an association between shorter patient survival as well as increased and decreased immunoreactivity, of the VEGFA and GLI1, respectively. The aforementioned findings suggest that the expression pattern of SHH, GLI1 and VEGFA demonstrates prognostic potential in KIRC. [ABSTRACT FROM AUTHOR]