Diagnosis of autoimmune bullous dermatoses: Comparative analysis of immunohistochemical staining using C4d, C3d, IgG, and IgG4 in lesional tissues and perilesional frozen skin samples.

Immunohistochemical staining with immunoglobulins and complements may aid the diagnosis of patients whose clinical and histological findings are consistent with autoimmune bullous dermatoses (AIBD). We aimed to investigate the diagnostic value of immunohistochemical markers in lesional biopsy and perilesional frozen samples in AIBD. We included 136 cases from whom lesional biopsies and perilesional samples for direct immunofluorescence (DIF) examination were collected with a preliminary diagnosis of AIBD between January 2019 and January 2023. All diagnoses were reconfirmed by evaluating the clinical, histopathological, and serological findings and DIF results (C3, IgG, IgA, or IgM positivity compatible with the clinical diagnosis) altogether, although DIF results were considered a priority. After confirming the diagnoses, the samples were categorized as AIBD or the others. The perilesional tissues obtained for DIF simultaneously with skin biopsy and stored at -80 °C were thawed, and FFPE tissues were prepared. We performed immunohistochemical staining (C4d, C3d, IgG, and IgG4) on FFPE tissues of both lesional and perilesional samples. Strong, linear, or granular staining patterns at the dermoepidermal junction or the intraepidermal blistering space were considered positive in line with the diagnosis of the case. Cases other than AIBD were used as negative control tissues to assess the specificity of immunohistochemical markers. Of the 136 cases, 52 were diagnosed with AIBD. In lesional samples, the sensitivity of C4d, C3d, IgG, and IgG4 was 80.6 %, 69.4 %, 75 %, and 5.7 % with corresponding specificity of 100 %, 98.7 %, 89.6 %, and 97.4 %, respectively in pemphigoid diseases compared to a sensitivity of 18.2 %, 9.1 %, 70 %, and 9.1 % and specificity of 98.7 %, 100 %, 89.6 %, and 97.4 %, respectively in pemphigus diseases. In frozen samples, we detected expression in a limited number of cases. The sensitivity of C4d, C3d, IgG, and IgG4 was 8.7 %, 2.2 %, 19.4 %, and 2.2 %, with corresponding specificity of 100 %, 100 %, 98.5 %, and 98.6, respectively. There was a none to slight concordance rate between the IHC results of lesional tissues and perilesional frozen samples. Kappa coefficients for C4d, C3d, IgG, and IgG4 were 0.120 (P = 0.029), 0.111 (P = 0.050), 0.203 (P = 0.003), and - 0.15 (P = 0.846), respectively. Immunohistochemical staining with C4d, C3d, IgG, and IgG4 on biopsy samples collected from lesions may guide the diagnosis of AIBD, thereby eliminating the need for an additional biopsy and accelerating the diagnostic process.

Immunoexpression Patterns of Adhesion Molecules (E-cadherin, ß-catenin, CD56) and Cytokeratins (CK19, CK20, HMWCK, CAM5.2) During Hair Development in Human Fetuses Compared With Adults.

ABSTRACT: Abnormalities in the expression of cytokeratins or adhesion molecules have been associated with hair disorders. The expression patterns of these molecules in the hair follicles of developing human fetuses are not obvious. We aimed to investigate the expression patterns of some cytokeratins and adhesion molecules in the hair follicle of human fetuses and compared them with adults. Forty-eight fetuses of >16 gestational weeks and 22 adult cases with total excisions of benign nevi or cysts were enrolled. The skin samples were taken from both the scalp and back of the fetuses. The histopathologically normal skin areas were evaluated in adults. CK19, CK20, CAM5.2, high-molecular-weight cytokeratin, E-cadherin, ß-catenin, and CD56 immunohistochemical stainings were performed. In the fetus group, the staining scores declined in the third trimester but elevated and reached the highest level in adults, except for CD56, which did not stain any adult samples. All stainings were mostly observed in the outer root sheath, except CD56 that stained the perifollicular dermal sheath only in fetuses. E-cadherin, ß-catenin, and high-molecular-weight cytokeratin strongly and diffusely stained all adult samples. CAM5.2 and CK19 scores were correlated in fetuses (scalp scores: r s = 0.405, P = 0.004; back scores: r s = 0.422, P = 0.003) and adults (back scores: r s = 0.562, P = 0.046). CD56 negativity indicated the immune-privilege feature of adult hair follicles. As CK19, CAM5.2 may be used to find the regions of stem cells or transient amplifying cells.

The effect of papaverine on tendon healing and adhesion in rats following Achilles tendon repair.

OBJECTIVES: The study aimed to examine the histopathological and biomechanical effects of papaverine administered intraperitoneally and locally on Achilles tendon healing in a rat model. MATERIALS AND METHODS: Forty-eight adult male Sprague-Dawley rats (range, 300 to 400 g) were used in this study conducted between October and November 2022. The rats were divided into three groups, with each group further subdivided into two for sacrifice on either the 15th (early period) or 30th (late period) day after surgery. The first (control) group received no treatment following Achilles tendon repair, while papaverine was intraperitoneally administered every other day for 10 days in the second group and locally in the third group after surgery. On the 15th and 30th days, the rats were sacrificed, and their Achilles tendons were subjected to biomechanical testing and histopathological evaluation. RESULTS: Histopathologically, there were no significant differences among the groups on the 15th day. However, on the 30th day, the locally applied papaverine group exhibited superior histopathological outcomes compared to the control group (p<0.05). Concerning the highest tensile strength values before rupture, the biomechanical assessment showed that the group receiving local papaverine treatment in the early period and both the group with systemic papaverine treatment and the one with local papaverine treatment in the late period displayed a statistically significant advantage compared to the control group (p<0.05). CONCLUSION: Locally administered papaverine has positive biomechanical effects in the early period and exhibits a positive correlation both histopathologically and biomechanically in the late period. Novel therapeutic options may be provided for patients through these findings.

Histopathological and histomorphometric investigation of the effects of different irrigation solutions on Achilles tendon healing in rats.

OBJECTIVES: This study aimed to examine the effect of irrigation fluids containing povidone-iodine (PVP-I), rifampicin (RF), and chlorhexidine gluconate (CHG) used during surgery on healing on a rat Achilles tendon model. MATERIALS AND METHODS: Twenty-eight male Sprague-Dawley rats (range, 300 to 400 g) were used in the experiment carried out between November 2022 and December 2022. The rats were divided into PVP-I, RF, CHG, and control groups, with seven rats in each group. Following the tenotomy and repair of the right Achilles tendon, the surgical site was irrigated using PVP-I, RF, CHG, or normal saline (the control group) for 2 min. All rats were sacrificed on the 21st postoperative day. The samples were evaluated histomorphometrically using the scoring system modified by Svensson, Soslowsky, and Cook and histopathologically using the Bonar and Movin classifications. RESULTS: The RF group gave better results in all three scoring systems compared to the control, PVP-I, and CHG groups (p=0.008, p=0.002, and p=0.006, respectively). Cellularity, rounding, and tenocyte morphology showed a significant difference in favor of the RF group (p=0.004). While the distribution of ground substance glycosaminoglycans showed a significant difference in favor of the RF group, there was no significant difference among the other groups (p=0.22). CONCLUSION: Irrigation solutions containing PVP-I, RF, or CHG show no negative effect on Achilles tendon healing. Moreover, the findings suggest that RF irrigation can accelerate the healing process.

Histopathological and molecular findings in 98 cases of endometrial carcinoma: MMR, p53 and next generation sequencing.

Gynecological malignancies arise from hereditary and somatic mutations, transcriptional aberrations, and genomic alterations influenced by epigenetic factors. This study aims to identify the mutations and their frequency in endometrial carcinomas (EC), and furthermore, to determine the relationship of these mutations with histopathological and immunohistochemical (IHC) parameters. The study was carried out in a retrospective cohort of 98 patients who received treatment upon being diagnosed with EC at a tertiary university hospital in Turkey between 2016 and 2021. The NGS-DNA tumor panel containing 29 genes was used in the study. NGS data of the cases were obtained from state of the evidence Tier 1 and 2 mutations. The relationship of patients' next generation sequencing (NGS) DNA panel results with histopathological parameters and IHC results (MLH1, MSH2, PMS2, MSH6 and p53) were evaluated. In 59 of the 98 cases, mutations were detected in at least one gene investigated with the NGS DNA panel. The most common somatic mutations in endometrial carcinoma were PIK3CA (33.6%), CTNNB1 (16.3%), KRAS (12.2%) and FGFR2 (11.2%) in this cohort. Abnormal p53 was detected by IHC in 15 out of 75 (20%) cases. Loss of expression was observed in at least one mismatch repair (MMR) protein by means of IHC in 39 out of 72 (39.8%) cases. Metastasis was found in 14 out of 82 (14.3%) patients who underwent lymph node dissection. FGFR2 mutation was more common in the group with lymph node metastasis compared to those without metastasis (p = 0.02). We report the mutational landscape of EC in a tertiary referral hospital in northwestern Turkey. Although our data are very limited, we think that the FGFR2 mutation may be associated with lymph node metastasis, but studies with larger patient numbers and longer follow-up periods are needed.

Effect of HER2 and Fascin expression on muscle-invasive bladder cancers: Classification by basaloid and luminal phenotypes.

Purpose: The present study aims to identify basaloid and luminal molecular groups and the p53-like sub-group, which is a sub-group of the luminal group, using a specific immunohistochemical panel and investigate human epithelial growth factor receptor 2 (HER2)/Neu and Fascin expression in these groups to analyze their relationship with clinicopathological features and prognosis in a cohort of cases with muscle-invasive urothelial bladder carcinoma (MIBC). Material and Methods: An immunohistochemical panel that included GATA-3, CK20, CD44, and CK5/6 was used to identify molecular sub-groups based on expression in 44 cases of MIBC. HER2/Neu and Fascin expression in basal, luminal, and p53-like groups and the relationship with clinicopathological features and prognosis were investigated. Results: The distribution of the molecular sub-groups determined by immunohistochemistry was as follows: 23 luminal cases (52.3%), 16 basal cases (36.4%), and 5 (11.4%) p53-like cases. There was a statistically significant difference in tumor size across the groups, with the greatest size in the p53-like group (p = 0.001). A statistically significant difference was observed in HER2/Neu expression between the molecular sub-groups (p = 0.017). Comparison of survival and HER2/Neu scores revealed shorter survival in patients with an HER2/Neu score of 3 + compared to those with scores of 0, 1+, and 2+ (p = 0.109). Fascin immunoreactivity was more common in the p53-like and basal groups compared to the luminal group (p = 0.036). Conclusion: Despite the limited number of cases in the MIBC group, our results support that HER2/Neu expression in the luminal sub-group and Fascin expression in basal and p53-like groups may be used as a negative prognostic marker. Multi-center studies that include large case series are warranted in this field.

Histiocytic Sarcoma with Invasive Urothelial Carcinoma of Urinary Bladder.

We report a rare case of histiocytic sarcoma in association with invasive urothelial carcinoma involving the urinary bladder. A 67-year male patient, who presented with a complaint of macroscopic hematuria, was found to have a mass on urinary system ultrasonography. Abdominal magnetic resonance imaging was performed to evaluate the mass, which showed a 52×24 mm mass on the posterior wall of the bladder. The cystoscopic examination revealed two suspicious areas in close proximity to one another with solid-papillary character. The tissue samples were collected by means of transurethral resection. The evaluation of these samples revealed two distinct neoplastic patterns. The areas of invasive urothelial carcinoma infiltrating lamina propria were noted on the surface in addition to diffuse sheets of large cells with hyperchromatic nuclei and ample clear cytoplasm, with a patternless pattern among small lymphocytes in myxoid background in the lamina propria. The positive reactions were observed in these areas with CD45, fascin, and CD68, a histiocytic marker. The histopathological diagnosis was histiocytic sarcoma in combination with invasive urothelial carcinoma. Histiocytic sarcoma may mimic several other malignant lesions, and only immunohistochemistry can identify this tumour to allow correct treatment. We present this rare case to emphasise that this phenomenon should be considered in unusual tumors and sites. Key Words: Urinary bladder, Histiocytic sarcoma, Invasive urothelial carcinoma.

Histopathological and immunohistochemical investigation of the local and systemic effects of tranexamic acid on the healing of the Achilles tendon in rats.

OBJECTIVES: This study aims to compare the effects of systemic and local applications of tranexamic acid (TXA) on tendon healing using a rat Achilles tendon injury model. PATIENTS AND METHODS: Thirty-six adult male albino Wistar rats (aging 3-4 months; weighing 350 to 400 g) were used in this study conducted between December 2019 and January 2020. The Achilles tendon was performed bilateral tenotomy and surgically repaired. Postoperatively, 1 mL of TXA was administered to each leg locally in the local group, whereas 2 mL of TXA was intraperitoneally administered in the systemic group. The control group was left untreated. Half of the rats were sacrificed on Day 15 and the other half on Day 30. Tendon healing was evaluated with the Bonar and the Movin scoring systems and immunohistochemical methods. RESULTS: The systemic group had the highest Bonar and Movin scores on Day 15. All groups exhibited tendon healing on Day 30, with no significant differences among the groups. The tenocyte morphology was found to be more impaired in both TXA groups on Day 30 (p=0.013). Ground substance scores were lower in the systemic group on Day 30 (p=0.028). The fiber structure and arrangement scores were higher in the systemic group on Day 15 (p=0.007 and p=0.032). Immunohistochemical analyses showed that galectin-3 values exhibited a significant difference in all groups on Day 30 (p=0.020). In all groups, it was determined that type I collagen values showed an increasing trend on Day 30, compared to the values on Day 15, whereas type III collagen values showed a decreasing trend. CONCLUSION: Our results demonstrated that local and systemic use of TXA does not impair tendon healing. Although advanced studies are needed, our study suggests that TXA application reduces the development of fibrosis.

The Effects of Fentanyl on Testicular Ischemia-Reperfusion Injury.

OBJECTIVE: Testicular torsion is a condition that often occurs as a result of the rotation of the spermatic cord in childhood and adolescence in men, manifests with acute pain and causes infertility in the future even if emergency intervention is performed. The aim of this study is to investigate the protective and preventive effects of fentanyl, a potent analgesic agent frequently used in anaesthesia practice, on testicular ischemia reperfusion injury, which manifests through acute pain. METHODS: A total of 16 adult male Wistar rats, weighing 200-250 g, were used in this study. They were divided into two groups, consisting of eight animals in each group. Torsion was created in all rats by rotating left testicles 720 clockwise on the day of the experiment. 3 mM of fentanyl was applied intraperitoneally 30 minutes before detorsion to the fentanyl group. Following an hour of ischemia, the left testicle was reinstated, and tissues were repaired according to their physiology. Following 24 hours of reperfusion, the animals were euthanised after taking left testes and blood samples. RESULTS: Fentanyl, administered prior to testicular detorsion, significantly suppressed germ cell damage in torsioned tissue, catalase activity and malondialdehyde levels in blood samples taken from the heart. No significant differences were observed in plasma total thiol concentration, histological score, Leydig cell counts, percentage of necrosis and tubule rupture. CONCLUSION: These findings show that fentanyl administered before detortion creates a protective effect by preventing testicular ischemia reperfusion injury leading to infertility in the future.

Vertical Bone Augmentation Using Bone Marrow-Derived Stem Cells: An In Vivo Study in the Rabbit Calvaria.

PURPOSE: To evaluate the bone regeneration capacity of bone marrow-derived stem cells (BMSCs) in vertical guided augmentation of bone tissue. MATERIAL AND METHODS: The calvaria of 20 rabbits were vertically augmented with autogenous bone graft (ABG); collagen/beta-tricalcium phosphate (ß-TCP) linked scaffold transplanted with 15 × 10 BMSCs; or scaffold alone (control). The augmentation materials were covered with stainless steel domes. BMSCs were isolated with Ficoll-Paque technique and applied directly without in vitro expansion. The newly formed bone was evaluated using radiodensitometric, histomorphometric, histological, and micro-computed tomographic (micro-CT) analyses after a 12-week healing period. The data excluding micro-CT assessments were compared statistically. RESULTS: Radiodensitometric and bone volume parameters demonstrated increased bone formation in both BMSC group and ABG group compared with control group (P < 0.01), but difference between the BMSC and ABG groups was not significant (P > 0.05). The mean histological scores for the BMSC, ABG, and control groups were 7.44 ± 1.03, 8.44 ± 0.81, and 6.00 ± 1.10, respectively, indicating significant difference among the groups (P < 0.05). CONCLUSION: BMSCs delivered with a collagen/ß-TCP linked scaffold can provide improved new bone formation that is comparable with autogenous bone block graft through vertical guided bone regeneration technique.

Risk factors for occupational brucellosis among veterinary personnel in Turkey.

Veterinarians and veterinary technicians are at risk for occupational brucellosis. We described the risk factors of occupational brucellosis among veterinary personnel in Turkey. A multicenter retrospective survey was performed among veterinary personnel who were actively working in the field. Of 712 veterinary personnel, 84 (11.8%) had occupational brucellosis. The median number of years since graduation was 7 (interquartile ranges [IQR], 4-11) years in the occupational brucellosis group, whereas this number was 9 (IQR, 4-16) years in the non-brucellosis group (p<0.001). In multivariable analysis, working in the private sector (odds ratio [OR], 2.8; 95% confidence interval [95% CI], 1.55-5.28, p=0.001), being male (OR, 4.5; 95% CI, 1.05-18.84, p=0.041), number of performed deliveries (OR, 1.01; 95% CI, 1.002-1.02, p=0.014), and injury during Brucella vaccine administration (OR, 5.4; 95% CI, 3.16-9.3, p<0.001) were found to be risk factors for occupational brucellosis. We suggest that all veterinary personnel should be trained on brucellosis and the importance of using personal protective equipment in order to avoid this infection.