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OBJECTIVE: This cross-sectional study was intended to assess the association between immunohistochemical analysis of p16 and fragile histidine triad (FHIT) and the presence of precancerous cervical lesions. MATERIALS AND METHODS: Women seen at Pérola Byington Hospital, Sao Paulo, Brazil, with histologically confirmed cervicitis (n = 31), cervical intraepithelial neoplasia (CIN) 1 (n = 30), CIN 2,3 (n = 30), and cervical cancer (n = 7) had also cervical material collected for liquid-based cytology, human papillomavirus Hybrid Capture 2 (HC2) test, and p16 and FHIT immunohistochemical reactions. RESULTS: p16 and FHIT reactions were scored as the following: <1%, 1% to 5%, >5% to 25%, and >25%. Receiver operating curve analysis was used to select p16 and FHIT score cutoffs for further categorical analyses. All but one of the 37 CIN 2,3/cancer cases had a p16 score of greater than 1% to 5%. Among the 61 cervicitis/CIN 1 cases, 46 (75%) had a p16 score lower than 1% to 5%. In contrast, no association of FHIT expression and severity of cervical lesions could be demonstrated in this data set. Receiver operating curve analyses suggested the score of 1% to 5% for p16 as the cutoff that best discriminates CIN 2,3/cancer from cervicitis/CIN 1. No cutoff for FHIT scores could be suggested with data set. CONCLUSIONS: p16, but not FHIT expression, has the potential to be used as complementary diagnostic tool to investigate human papillomavirus-induced cervical lesions, if these results are confirmed in larger studies. [ABSTRACT FROM AUTHOR]

Objective: To optimize the protocols of human papillomavirus (HPV) detection in clinical samples, we used polymerase chain reaction (PCR)-based techniques in paraffin-embedded tissue sections and compared the results with those obtained with PCR and Hybrid Capture II (HC2) performed in liquid-based cytology (LBC) preservation medium.Materials and Methods: Forty-five consecutive cervical biopsy specimens were taken from women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, São Paulo, Brazil, during 2003 and 2004. The biopsy specimens were analyzed for HPV by a modified GP5+/GP6+ PCR protocol, and the results were compared with those obtained by PGMY PCR and HC2 in samples collected in LBC preservation medium.Results: beta-Globin was detected in 100% of the multiplex PCR system from LBC samples and 66.7% with PCO4+/PCO3+ PCR in biopsy specimens. Of the three methods, PGMY PCR system and HC2 were equally effective in detecting HPV; both detected 13 cases in 45 samples (28.9%). The GP5+/GP6+ PCR applied in biopsy specimens showed a 20% HPV detection rate (9/45).Conclusions: Our PCR protocols worked reasonably well and allowed us to compare the three molecular methods with histological and cytological findings. The reproducibility of the results makes the technique applicable in archival materials. [ABSTRACT FROM AUTHOR]

Advancements in cutting-edge molecular profiling techniques, such as next-generation sequencing and bioinformatic analytic tools, have allowed researchers to examine tumour biology in detail and stratify patients based on factors linked with clinical outcome and response to therapy. This manuscript highlights the most relevant prognostic and predictive biomarkers in kidney, bladder, prostate and testicular cancers with recognised impact in clinical practice. In bladder and prostate cancer, new genetic acquisitions concerning the biology of tumours have modified the therapeutic scenario and led to the approval of target directed therapies, increasing the quality of patient care. Thus, it has become of paramount importance to choose adequate molecular tests, i.e., FGFR screening for urothelial cancer and BRCA1-2 alterations for prostate cancer, to guide the treatment plan for patients. While no tissue or blood-based biomarkers are currently used in routine clinical practice for renal cell carcinoma and testicular cancers, the field is quickly expanding. In kidney tumours, gene expression signatures might be the key to identify patients who will respond better to immunotherapy or anti-angiogenic drugs. In testicular germ cell tumours, the use of microRNA has outperformed conventional serum biomarkers in the diagnosis of primary tumours, prediction of chemoresistance, follow-up monitoring, and relapse prediction., (Copyright © 2023 Royal College of Pathologists of Australasia. Published by Elsevier B.V. All rights reserved.)

A wide variety of diseases present with intracranial lesions. In this case report, a 67-year-old man initially presented to an outside hospital with nausea, headache, and ataxia and was found to have multiple intracranial lesions. Diagnostic workup was ultimately unrevealing, and his condition improved after a course of steroids and antibiotics. Unfortunately, symptoms returned 3 months later. MRI of the brain revealed progression of his intracranial lesions. This case highlights a diagnostic approach and general management strategy for patients presenting with undifferentiated intracranial pathology. A final diagnosis is ultimately reached and raises further discussion., (© 2023 American Academy of Neurology.)

Age is an important prognostic factor in melanoma; notably, elderly patients tend to present with advanced stage skin melanoma (SM) and worse outcome. Moreover, SM is an immunogenic cancer, and its interaction with the aging immune system could have an effect on biologic behaviour of this disease. Tumour-infiltrating lymphocytes (TILs) could represent the host response in SM; it has been shown that higher grade of TILs is associated with better survival. Moreover, programmed death ligand 1 (PD-L1) and cyclooxygenase-2 (COX-2) are potential markers of host immune response and inflammation. We retrospectively reviewed 113 consecutive cases of early-stage SM that occurred in patients aged greater than or equal to 65 years at the time of diagnosis, followed between January 2010 and March 2014 at the University and General Hospital of Udine, Italy. The aim of this study was to evaluate TILs grade, PD-L1 expression on TILs and tumour expression of PD-L1 and COX-2 and their prognostic value in elderly patients with early SM. A better disease-free survival as well as melanoma-specific survival (MSS) was significantly associated with TILs [hazard ratios (HR): 0.41, 95% confidence interval (CI): 0.20-0.84, P=0.02 and HR: 0.37, 95% CI: 0.17-0.82, P=0.01, respectively]. PD-L1 positivity on TILs was associated with a better MSS (HR: 0.41, 95% CI: 0.17-0.97, P=0.04). Moreover, among patients with TILs, those showing COX-2 positivity on tumour cells and no PD-L1 expression on TILs had a worse disease-free survival and MSS (HR: 5.18, 95% CI: 1.33-20.23, P=0.018; HR: 6.21, 95% CI: 1.20-32.24, P=0.03; respectively). Immune and inflammatory markers deserve further investigation in aging patients with melanoma.

Primary adenocarcinomas of the gastrointestinal tract showing a diffuse thyroid transcription factor-1 (TTF-1)-positive immunostaining are extremely uncommon and this finding in the biopsy specimen could yield to misleading diagnosis of a metastatic lung tumor. We report the clinical and pathological features of 2 cases, occurring in a 62-year-old woman and in a 66-year-old man. The first one is a gastric adenocarcinoma with liver metastasis; the other one is a gallbladder adenocarcinoma discovered during the screening for the colorectal cancer. Both the neoplasms showed a diffuse TTF-1 positivity and both the patients did not have any evidence of lung cancer. Neither a gastric adenocrcinoma with diffuse TTF-1 expression nor a TTF-1-positive gallbladder adenocarcinoma has been described before.

A platform composed of a microwave (MW)-assisted tissue-processing system and an automated embedding system has been recently introduced in pathology laboratories. Needle core biopsy (NCB) is an established, highly accurate method for diagnosing breast lesions and for providing important pathologic, predictive, and prognostic information such as biomarker expression in case of breast carcinoma. The aim of this study was to evaluate whether breast NCBs processed with the MW-assisted tissue-processing system and automatically embedded show good-quality histology preparations and whether they are suitable for the assessment of estrogen receptor (ER), progesterone receptor (PR), Ki-67, and HER2 in breast carcinoma. A series of 233 consecutive breast NCBs processed by both conventional and MW-assisted tissue-processing systems was included in this study. The histomorphologic and immunohistochemical quality, as well as the results of the evaluation of the biomarkers, were compared-the conventional processing method being the gold standard for comparison. The quality of hematoxylin-eosin and immunohistochemical tissue sections provided by the new system is comparable to that obtained after the conventional processing method. Moreover, in breast carcinomas, a perfect agreement between the paired tissues when evaluating ER and PR status (Cohen κ = 1) and a very good agreement when evaluating Ki-67 (κ = 0.91) and HER2 (κ = 0.93) have been found. In conclusion, applying strict criteria in tissue-handling steps, breast NCB can be processed and automatically embedded with these platforms. The diagnosability and the evaluation of the main prognostic and predictive biomarkers have been proved to be reliable.

Several studies have reported an increase in vascular structures in malignant melanoma. Neovascularization can be enhanced by several factors. Among them, thymidine phosphorylase (TP) and cyclooxygenase-2 (COX-2) have been reported to play a role. The expressions of TP and COX-2 were evaluated trough immunohistochemistry in a series of 78 primary cutaneous melanomas diagnosed between 2000 and 2004. The expressions of TP and COX-2 through mRNA and western blot analysis were also evaluated in several melanoma cell lines. TP expression and COX-2 expression were considered positive in 25 cases (32%) and 22 cases (28.2%), respectively. TP-positive melanomas showed a lower mitotic rate (P=0.008), smaller thickness (P=0.01), and absence of lymphovascular invasion (P=0.04). COX-2-positive melanomas showed a higher mitotic rate (P=0.01) and higher thickness (P=0.03). COX-2 expression was associated with reduced disease-free survival (P=0.01). COX-2-positive cases showed a trend toward reduced survival, whereas TP was not correlated with overall survival. COX-2 expression was detected in four of 11 melanoma cell lines both by mRNA and by western blot analysis. Our data show that TP expression is associated with more favorable prognostic factors (such as thin melanoma, low mitotic count, and absence of lymphovascular invasion), whereas COX-2 expression is associated with poor prognostic factors (thicker melanoma and high mitotic count)., (© 2013 Wolters Kluwer Health | Lippincott Williams & Wilkins.)

The aim of the present study is to evaluate the clinical and biological factors (including markers of angiogenesis) as potential predictors of prognosis and benefit from metronomic therapy in patients with advanced breast cancer (ABC). Recent data suggest antiangiogenic activity of metronomic therapy. The study population included 62 patients with pretreated ABC who received cyclophosphamide and methotrexate orally. Tumour samples were analysed by immunohistochemistry for HER2, Ki-67, thymidine phosphorylase (TP), vascular endothelial growth factor and vascular endothelial growth factor receptor. The results from immunohistochemical analysis and clinico-pathological variables were studied to test their potential association with benefit from metronomic therapy. The median overall survival, progression-free survival and survival postprogression were 7.1 (range 0.2-38.3), 2.6 (range 0.2-28.9) and 3 (range 0-34.2) months, respectively. Among the clinical variables, age, performance status and previous therapy with taxanes were significantly associated with outcomes. Among the molecular markers, TP was found to be associated with progression-free survival. Metronomic therapy is an effective choice for ABC. Young women with a more indolent disease had the greatest benefit from this treatment. TP tumour expression might aid decision making but these findings must be confirmed in larger prospective, properly designed studies.

This study investigated if tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) immunohistochemical expression in human temporomandibular joint (TMJ)-degenerated disks correlates to the degree of tissue damage to elucidate the possible involvement of this apoptotic pathway in TMJ disk degeneration. Twenty-one TMJ displaced disk from 12 patients were affected by anterior disk displacement with reduction and 9 by anterior disk displacement without reduction processed immunohistochemically with TRAIL antibody. Histopathologic grading of the disk degeneration was carried out in each specimen. The mean histopathologic score of the TMJ degenerated disks was 4.77±1.26 (minimum, 2; maximum, 7). Immunolabeling for TRAIL was detected in the cytoplasm of the TMJ disk cells in every sample, although with different patterns of reactivity. The degree of TRAIL immunostaining was correlated to the histopathologic degeneration score obtained from the sample (Spearman ρ=0.617). Therefore, cell loss due to the involvement of TRAIL apoptotic pathway seems, in part, responsible for TMJ disk degeneration.

In this study, we hypothesized that caspase 3, which plays a central role in the execution phase of cell apoptosis, could be involved in limiting fatty degeneration of the temporomandibular joint (TMJ) disks and therefore inhibit the TMJ disk tissue from completely degenerating into fatty tissue. Therefore, caspase 3 immunohistochemical expression in human TMJ degenerated disks was studied. Fifty-nine degenerated TMJ disks were stained with Harry's hematoxylin, and they were then examined with light microscopy to detect any pathologic changes typical of fatty degeneration. Sections from the same TMJ disk were immunostained also by a polyclonal anti-caspase 3 antibody. On morphologic observations, 11 disks of 59 degenerated ones also presented a fatty infiltration. Immunostaining with caspase 3 antibody was detected on adipocytes in the cytoplasm as well as the nuclei. Our results sustain the hypothesis that fatty degeneration is limited by apoptosis, being adipocytes immunolabeled by caspase 3 antibody. Hence, apart from the several factors that can trigger degeneration changes in TMJ disk, their appearance, spread, and permanence, at least for fatty degeneration, seem to be influenced by apoptosis.

This study describes, microscopically, the pathologic findings correlated to malpositioning of the human temporomandibular joint disc. The specimens studied consisted of 21 discs removed from patients affected by temporomandibular joint arthropathy. The specimens were cut longitudinally and were fixed overnight in 10% neutral-buffered formalin. They were embedded in paraffin with anatomic orientation preserved. Three- to 4-microm thick sections were cut according to routine procedures and mounted on slides. Then they were stained with Goldner-modified-Mallory staining used for morphologic study at light microscopy. Abnormal collagen fiber arrangements, fragmentation of collagen fibrils, new vessel formation, meniscal tears, mucoid degeneration of the disc matrix, chondrocyte-like cells proliferation, sometimes with clonal aggregation, and hyalinization were detected in temporomandibular joint disc specimens with internal derangement, although with a different extent. From our observations, it seems that in most cases, the temporomandibular joint disc responded to internal derangement without reduction with a fragmentation of collagen fibers because hyalinization of disc matrix occurred very rarely. Moreover, disc derangement without reduction is characterized also by a change in cell population phenotype.

We have investigated the immunohistochemical expression of thymidine phosphorylase before and after the administration of anthracycline, and/or anthracycline/taxane-based preoperative chemotherapy in a consecutive series of 55 patients with primary operable breast cancer. Pretreatment, large core breast biopsies and the corresponding surgical samples were retrospectively evaluated for thymidine phosphorylase immunoreactivity. Immunohistochemical expression was evaluated on tumor cells (nuclear and cytoplasmic staining) and on stromal cells (cytoplasmic staining). The cytoplasmic expression of thymidine phosphorylase was enhanced in the tumor cells after treatment (P=0.04). An increase in thymidine phosphorylase cytoplasmic tumor expression was observed in 33% (95% confidence interval: 19-50%) of patients after preoperative chemotherapy (P=0.01). No statistically significant nuclear staining changes were observed in response to treatment. Similarly, no significant changes of the enzyme expression were seen in stromal cells. This study provides further evidence that, at least in breast cancer, thymidine phosphorylase is upregulated after anthracycline and/or taxane-containing chemotherapy. Accordingly, it supports the hypothesis of a synergistic effect between thymidine phosphorylase-modulating and thymidine phosphorylase-targeting anticancer agents. Translational studies, specifically designed on the basis of this rationale, are eagerly waited.

This study was designed to evaluate whether Hybrid Capture II (HC2) test alone refer women to colposcopy as appropriately as DNA Papanicolaou (Pap) test, in the context of a high-risk group of women using the recently validated DNACitoliq LBC system. Women with suspected cervical disease were included in this cross-sectional study at a tertiary center in São Paulo, Brazil, for further workup. All women had cervical material collected for LBC and HC2 for high-risk human papillomavirus (hrHPV)-DNA test. Irrespective of cytology and HC2 results, colposcopy, and cervical biopsy when applicable, was systematically performed. All tests were performed blindly. Sensitivity, specificity, positive and negative predictive values, and overall accuracy of both methods were computed in relation to histology. A total of 1,080 women were included: 36.4% (393/1080) had ACUS+, 10.2% (110/1080) were high-grade squamous intraepithelial lesions (HSIL) or cancer. Mean age was 33.5 years. All women underwent colposcopy, and cervical biopsies were performed in 38.4% (415/1080): 33% (137/415) of the biopsies were negative, 14.4% (155/415) were low-grade squamous intraepithelial lesions (LSIL), 10.7% (116/415) were HSIL, and 0.6% (7/415) were cancer. HC2 sensitivity to diagnose biopsy-proven HSIL was 100%. Because all HSIL cases had a positive HC2 test, sensitivity could not be improved by adding LBC. Specificity and positive and negative predictive values of DNA Pap were not significantly different from HC2 test alone when considering LSIL+ histology as "gold standard" and HSIL+ histology. As a screening strategy for women with high-risk for cervical cancer, DNA Pap test does not seem to add substantially to HC2 alone in terms of appropriately referring to colposcopy.

This study was designed to analyze the cross-sectional comparison of the p16 and Ki-67 immunocytochemical expression in negative and equivocal (atypical squamous cells of undetermined significance (ASC-US)) liquid-based cytology (LBC) samples testing positive for high-risk human papillomavirus (HPV) types with HC2 assay or polymerase-chain reaction (PCR). A series of 199 consecutive LBC specimens derived from the same number of women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, Sao Paulo, were analyzed using immunocytochemistry for expression of p16 and Ki-67 in negative and equivocal LBC samples testing positive for high-risk HPV types with hybrid capture II test (HC2) or PCR. All patients with at least one test positive (cytology, PCR, and/or HC2) were followed each 6 months for 3 years. The follow-up procedure consisted of visual examination, colposcopic inspection, cytology, and HC2 assay. Among the negative cytologic samples, 101 were HPV-positive and 55 HPV-negative. Of the HPV-positive group, 59 of 101 cases (58.4%) were positive for both p16 and Ki67 immunostaining, and 17 of 101 (16.8%) were negative for both. The proportion of Ki-67-positivity increased almost in parallel with the increasing grade of p16-positivity (p = 0.0001 for linear trend). In the HPV-negative group, both markers were negative in 41 of 55 cases (74.5%), and no statistical relationship was observed between the two markers (Pearson, p = 0.595). HPV-positive ASC-US samples demonstrated a simultaneous positive immunoreaction for p16 and Ki67 in 11 of 16 cases (68.7%), whereas 3 (18.7%) were concurrently negative. The relationship between the two markers was of borderline significance (Pearson, p = 0.053), but no linear relationship was found between the graded p16 and Ki-67 expression (p = 0.065 for linear trend). In the HPV-negative ASC-US group, there was no statistical association between the graded p16 and Ki-67 positivity (Pearson, p = 0.281). After 36 months of follow-up of the ASC-US patients, 6 women still displayed ASC-US smear, of which 4 of 6 were HPV-positive and expressed both p16 and Ki-67 markers. Two of 43 ASC-US smears had high-grade squamous intraepithelial lesions diagnosed (4.6%), and 1 had low-grade squamous intraepithelial lesion (2.3%). All of those were positive for HPV, p16 and Ki-67. Patients with ASC-US diagnosis and positive high-risk HPV status and positive for p16 Ki67 should be carefully observed to exclude occurrence of a squamous intraepithelial lesion. The combination of these two markers can be a useful implement for management of women with equivocal cytology.

Heat shock proteins (HSPs) are believed to represent a cellular stress response mechanism that protects intracellular proteins from damaging events. Some studies have demonstrated an enhanced expression of large-molecular-weight HSPs in diseased systematic joints. Small heat shock proteins, and among these HSP27, have been studied to a lesser extent. HSP27 has cytoprotective and biosynthetic functions within chondrocytes, and it is an estrogen-associated protein that is under hormonal modulation. To improve understanding at a molecular level of the pathophysiology of certain temporomandibular joint disorders, the authors carried out this immunohistochemical study to assess the presence of HSP27 in human TMJ discs. Twelve adult human TMJ discs (10 diseased and 2 healthy discs) and 5 TMJ fetal human discs were used in this study. Adult discs and TMJ tissues of human fetuses were fixed in 10% buffered formalin. Samples were then processed for histologic examination. Sections were immunohistochemically stained using the streptavidin-biotin detection method. No reaction product for HSP27 in the discs of fetuses was noted. HSP27 was weakly expressed in normal TMJ discs and highly up-regulated in discs of patients showing new vessel formation and chondroid metaplasia. Any correlation between gender and HSP27 was found in the sample, being the up-regulation of HSP27 related mostly to major histopathological changes. This different pattern of HSP27 immunostaining in human TMJ discs detected in the authors' specimens suggests that the expression of this small HSP is functionally modulated. In fact HSP27 up-regulates in internal derangement specimens with major histopathological changes; on the other hand, it is not expressed or only weakly expressed in TMJ discs of fetuses and normal TMJ discs.

Cardiovascular disease is delayed and less common in women than in men. Myocyte death occurs in heart failure, but only apoptosis has been documented; the role of myocyte necrosis is unknown. Therefore, we tested whether necrosis is as important as apoptosis and whether myocyte death is lower in women than in men with heart failure. Molecular probes were used to measure the magnitude of myocyte necrosis and apoptosis in 7 women and 12 men undergoing transplantation for cardiac failure. Myocyte necrosis was evaluated by detection of DNA damage with blunt end fragments, whereas apoptosis was assessed by the identification of double-strand DNA cleavage with single base or longer 3' overhangs. An identical analysis of these forms of cell death was performed in control myocardium. Heart failure showed levels of myocyte necrosis 7-fold greater than apoptosis in patients of both sexes. However, cell death was 2-fold higher in men than in women. Heart failure resulted in a 13-fold and 27-fold increase in necrosis in women and men, respectively. Apoptosis increased 35-fold in women and 85-fold in men. The differences in cell death between women and men were confirmed by the electrophoretic pattern of DNA diffusion and laddering of isolated myocytes. The lower degree of cell death in women was associated with a longer duration of the myopathy, a later onset of cardiac decompensation, and a longer interval between heart failure and transplantation. In conclusion, myocyte necrosis and apoptosis affect the decompensated human heart; each contributes to the evolution of cardiac failure. However, the female heart is protected, at least in part, from necrotic and apoptotic death signals.