Flow-cytometry Assessment of DNA content and Immunophenotyping of Immune-cells in Lymph-node-specimens as a Potential Diagnostic Signature of Aggressiveness in B-Non-Hodgkin Lymphomas.

Flow-cytometry (FC) is a powerful tool that can assist in lymphoma diagnosis in lymph node (LN) specimens. Although lymphoma diagnosis and classification are mainly based on tumor cell characteristics, surrounding cells are less employed in this process. We retrospectively investigated alterations in the ploidy status, proliferative cell fraction (PF) and the percentages of surrounding immune cells in 62 consecutive LN specimens with B-Cell Non-Hodgkin Lymphoma (B-NHL) that were submitted for FC evaluation between 2019-2022. Compared with indolent B-NHLs, aggressive B-NHLs show increased DNA aneuploidy and PF, increased monocytes, immature-granulocytes, mature granulocytes, CD8+ T-cells, Double-Negative-T-cells and Double-Positive-T-cells, and decreased total CD45+ cells, total lymphocytes, CD4+ T-cells and CD4/CD8 ratio. Receiver operating characteristic analysis determined PF > 6.8% and immature-granulocytes > 0.9% as optimal cutoffs with highest specificity and sensitivity in differentiating aggressive and indolent B-NHLs. These findings further strength the diagnostic value of DNA content analysis by FC and suggest the utilization of tumor surrounding immune cells in NHL diagnosis and classification.

Coexisting Epithelioid Trophoblastic Tumor and Placental Site Trophoblastic Tumor During Asymptomatic Relapse: A Case Report and Literature Review.

Gestational trophoblastic neoplasms are a group of trophoblastic tumors that include choriocarcinoma (CC), epithelioid trophoblastic tumors (ETTs), and placental site trophoblastic tumors (PSTTs). Mixed gestational trophoblastic neoplasms include combinations of CCs with ETTs and/or PSTTs; combinations of ETTs and PSTTs have also been described. This report describes the case of a 49-yr-old female with mixed ETT and PSTT discovered due to menstrual delay and a positive beta-human chorionic gonadotropin in serum 11 yr after normal pregnancy; it is an asymptomatic recurrence of the neoplasm after 2 yr. Moreover, only the ETT recurred without evidence of PSTT by biopsy and without any increase in human chorionic gonadotropin levels, even though human chorionic gonadotropin was positive in the first onset of the disease. We also reviewed published English literature, which revealed that there are only 36 cases of mixed trophoblastic tumors to date, of which pure mixed ETT and PSTT were reported only in four cases including our case. The most common combination is CC admixed with an ETT (52%), followed by CC with PSTT in 30.5%. CC admixed with an ETT and/or PSTT account for 83% of the cases, of which pure mixed ETT and PSTT were reported only in 4 cases (11%). The rarity of this condition entails reporting of all cases to facilitate future research and clinical management.

Not All Bethesda 1 Thyroid Nodules Were Created Equal: Different B1 Subgroups.

OBJECTIVE: The Bethesda System for Reporting Thyroid Cytopathology is a uniform method used worldwide to report thyroid fine-needle aspiration (FNA) outcomes. This study focuses on the Nondiagnostic/Unsatisfactory category, designated as Bethesda1 (B1). The documented risk of malignancy for B1 nodules can vary significantly, implying this category is not homogenous and might be composed of different subtypes. Our hypothesis was that B1 subgroups (blood only, insufficient thyrocytes, cyst content) will vary in their malignancy rate. METHODS: The study design was observational and retrospective. The study population included 154 patients in the Galilee Medical Center who underwent FNA examination of the thyroid gland from 2013-2018 and had a B1 result. We looked at the final diagnosis of malignant or benign for patients who underwent surgery and calculated the malignancy rate for each subgroup. RESULTS: Malignancy rates were higher in the Blood subgroup than in the other subgroups, and higher in the Thyrocytes subgroup than in the Cyst subgroup (P < .05). All malignancies were papillary thyroid carcinomas. There was no significant difference in the malignancy rate when we further divided the B1 samples into 2 groups based on the presence of epithelial cells. Many repeat FNA tests resulted in a different B1 subgroup. CONCLUSION: The different malignancy rates suggest that individual management approaches should be considered for each B1 subgroup.

Differences in cytopathologist thyroid nodule malignancy rate.

OBJECTIVES: The accuracy of a cytological diagnosis obtained by fine needle aspiration is influenced by several factors including the technique used and the experience of both the aspirator as well as the cytologist. In this project we planned to evaluate the interobserver differences of thyroid nodule cytopathology in our medical centre. METHODS: The study was conducted using retrospective pathology reports from a single academic centre from August 2013 to September 2017. We compared the sensitivity, specificity, negative and positive predictive values, malignancy rates, and accuracy of two cytopathologists who evaluated thyroid nodules. RESULTS: We included 287 fine needle aspirations of thyroid nodules in the study. Approximately one fifth (18.5%) of patients had surgery and the rate of malignancy was 40%. There was a similar frequency of use of all thyroid Bethesda system (TBS) categories with the exception of TBS 3 (8.0% and 21.2%, P = .01). As a consequence, the malignancy rate was different in TBS 3 category (40% vs 17%, P = .545). CONCLUSIONS: There are interobserver differences in the evaluation of thyroid nodules. Clinicians should be aware of such differences because they affect the malignancy rate in each TBS category.

Review of a single institution's fine needle aspiration results for thyroid nodules: Initial observations and lessons for the future.

OBJECTIVE: Our objective was to evaluate the disease spectrum of thyroid cytopathology and correlation of the Bethesda reporting system with final histopathology in our medical centre. METHODS: This retrospective study was conducted from histopathology reports from Galilee Medical Center between August 2013 and September 2017. RESULTS: A total of 287 thyroid fine needle aspirations were included in the study. The majority (55.1%) of these were benign (B2). Surgery was performed on 53 cases and the total malignancy rate was 39.6%. Our study had a favourable accuracy rate of 70%. A B4 Bethesda category had a higher malignancy rate (50%) than previously reported. CONCLUSION: We found a higher malignancy rate (50%) based on the Bethesda B4 category of the fine needle aspirations in our series, yet it is based on a small sample. The differences in malignancy rates between centres have an important impact on clinical decisions.

A founder mutation in Vps37A causes autosomal recessive complex hereditary spastic paraparesis.

BACKGROUND: Members of two seemingly unrelated kindreds of Arab Moslem origin presented with pronounced early onset spastic paraparesis of upper and lower limbs, mild intellectual disability, kyphosis, pectus carinatum and hypertrichosis. METHODS: The authors performed neurological and developmental examinations on the affected individuals. The authors conducted whole genome linkage and haplotype analyses, followed by sequencing of candidate genes; RNA and protein expression studies; and finally proof of principle investigations on knockdown morpholino oligonucleotide injected zebrafish. RESULTS: The authors characterise a novel form of autosomal recessive complex hereditary spastic paraparesis (CHSP). MRI studies of brain and spinal cord were normal. Within a single significantly linked locus the authors ultimately identified a homozygous missense mutation c.1146A>T (p.K382N) in the vacuolar protein sorting 37A (Vps37A) gene, fully penetrant and segregating with the disease in both families. Mobility was significantly reduced in Vps37A knockdown morpholino oligonucleotide injected zebrafish, supporting the causal relationship between mutations in this gene and the phenotype described in the patients of this study. CONCLUSIONS: The authors provide evidence for the involvement of Vps37A, a member of the endosomal sorting complex required for transport (ESCRT) system, in upper motor neuron disease. The ESCRT system has been shown to play a central role in intracellular trafficking, in the maturation of multivesicular bodies and the sorting of ubiquitinated membrane proteins into internal luminal vesicles. Further investigation of mechanisms by which dysfunction of this gene causes CHSP will contribute to the understanding of intracellular trafficking of vesicles by the ESCRT machinery and its relevance to CHSP.

Nondiagnostic Test Result Rates Following Thyroid Extra-Fine-Needle Aspiration.

OBJECTIVES: The first Bethesda classification category of thyroid fine-needle aspiration biopsy (FNAB) is nondiagnostic (ND), which indicates that the specimen's cellularity is inadequate for evaluation. This work investigated the effect of needle diameter size on ND rates by comparing diagnostic outcomes of FNAB samples collected with 23-, 25-, and 27-gauge needles. METHODS: This was a retrospective analysis of samples collected from patients undergoing FNAB between 2018 and 2021. It was conducted in an otolaryngology department in a university teaching hospital. RESULTS: Of the 699 aspirations, 144, 335, and 220 were performed using 23-, 25-, and 27-gauge needles, respectively. ND rates increased significantly when using 27-gauge compared with 23- to 25-gauge needles (P = .002), and a significantly lower ND rate was found for the 25-gauge needle compared with the 27-gauge needle (P = .001). Furthermore, increased nodule size was associated with reduced ND rate (odds ratio, 0.801; 95% confidence interval, 0.691-0.929). CONCLUSIONS: The 25-gauge needles are superior to 27-gauge needles in reducing ND rates of thyroid nodule FNAB specimens. Future prospective studies should be performed to confirm these findings.

Effect of needle gauge on thyroid FNA diagnostic rate.

PURPOSE: Thyroid Bethesda classification system provides 6 diagnostic categories, the first being a sample deemed non-diagnostic or insufficient and requiring a subsequent second biopsy. Our objective was to evaluate differences in non-diagnostic fine needle aspiration (FNA) of thyroid nodules conducted with a 23-gauge(G) needle vs. those conducted with a 25 G needle. METHODS: Data from 298 aspiration procedures using either 23 G or 25 G needles were collected, including cytological findings, ultrasound characteristics and patient demographics. The samples were classified as diagnostic or non-diagnostic according to final cytology. RESULTS: There was no statistically significant difference between the 25 G and 23 G needles in terms of non-diagnostic rates (35.7%, 31.9%; p = 0.494). Nodules defined as cystic had higher non-diagnostic rates (p < 0.05). Older patients as well as cystic nodules were associated with a higher non-diagnostic rate (OR = 1.018, p = 0.047, OR = 13.533, p = 0.0001, respectively), while nodule size was associated with lower non-diagnostic rates (OR = 0.747, p = 0.017). CONCLUSIONS: The use of 25 G needle did not produce a lower non-diagnostic rate when compared to 23 G needle. Larger nodules might increase diagnostic rates, while older patients and cystic nodules are prone to inadequate samples. Patients and caregivers should be aware that FNA of small or cystic nodules as well as nodules in older patients may result in a higher non-diagnostic rate. Further research comparing other needles gauges should be conducted.

Hemorrhagic Malignant Pleural Effusion: Diagnosis, Survival Rate, and Response to Talc Pleurodesis.

Hemorrhagic malignant pleural effusion (HMPE) is diagnosed in 47-50% of all malignant pleural effusions (MPE). The aim of this study is to evaluate clinical, radiological, and morphological manifestations of HMPE and results of talc pleurodesis treatment. This is a retrospective review of the medical charts of 135 MPE patients which consists of HMPE group (42 patients) and simple MPE group (63 patients) (median age 67.9 years; 43 males, 62 females). In HMPE vs. simple MPE patients, pronounced dyspnea (100% vs. 88.9%, P = 0.024), chest pain (59.5% vs. 60.3%, P = 1), general deterioration (78.6% vs. 74.6%, P = 0.411) combined with large pleural effusion (81% vs. 50.8%, P = 0.001), and thickening of parietal pleura (73.8% vs. 68.3%, P = 0.349), all were more specific for HMPE. Cytological examination of HMPE showed more malignant pleural fluid cells (81% vs. 63.5%, P = 0.043). Histological examination revealed poorly differentiated types of tumors in 69.05% of HMPE (bronchogenic 33.33%, intestinal 16.67%, breast 14.3%) vs. 7.94% of simple MPE. In 19 HMPE vs. 0 simple MPE patients, thoracoscopy showed bleeding nodules (94.7%) on thickened parietal pleura (84.2%). Pleurodesis with talc by slurry (59%) and poudrage (41%) was less effective in HMPE than in simple MPE patients after 1 month (failed response; 33.3% vs. 21.6, P = 0.019), 3 months (42.9% vs. 25.7%, P = 0.017), and 6 months (42.9% vs. 21.7%, P = 0.035). Survival in HMPE was significantly lower (3.06 months vs. 5.37 months, P = 0.0005). HMPE has more severe clinical, laboratory, radiological, and endoscopic manifestations due to a more poorly differentiated malignant process. Talc pleurodesis was less effective in HMPE, and survival was poor.

Flow cytometry aneuploidy and cell cycle indexing as a possible tool for differentiating between CD10+ diffuse large B-cell lymphoma and follicular lymphoma.

BACKGROUND: Differential diagnosis between diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) becomes a challenge when adequate biopsy is unavailable. The present study aimed to investigate the applicability of DNA cell cycle analysis by flow cytometry (FC) for differentiating between CD10+ DLBCL and FL. METHODS: Data were collected from 57 specimens where CD5- /CD10+ /light chain restricted B cells were detected. DNA staining was performed using the Coulter DNA Prep Kit. Cell cycle fractions were evaluated by automatic analysis using the ModFit LT software. RESULTS: Histopathological analysis confirmed the diagnosis of CD10+ FL in 30 specimens (52.6%), CD10+ DLBCL in 24 specimens (42.1%), and CD10+ Burkitt lymphoma in 3 specimens (5.3%). A significantly higher rate of DNA aneuploidy was detected among CD10+ DLBCL than FL specimens (50 vs. 13.3% respectively, p = .003). Likewise, DNA index was significantly higher in CD10+ DLBCL relative to FL (1.26 ± 0.35 vs. 1.04 ± 0.16 respectively, p = .004). Notably, the proportion of cells in the S-phase and proliferative fraction was significantly higher in CD10+ DLBCL than in CD10+ FL (S-phase: 15.97 ± 13.94 vs. 4.43 ± 4.41 mean ± SD, respectively, p < .0001; proliferative fraction: 18.87 ± 15.17 vs. 5.78 ± 7.04 mean ± SD, respectively, p = .0001). Using a receiver operating characteristic analysis, optimal cutoffs for S-phase ≥7% and proliferative fraction ≥9% were determined. These values could be used to differentiate between CD10+ DLBCL and CD10+ FL. CONCLUSION: Including DNA cell cycle analysis in the FC lymphoma assessment panel may be of diagnostic value in differentiating between CD10+ DLBCL and FL when adequate biopsy is unavailable.

Complex Nabothian cysts: a diagnostic dilemma.

BACKGROUND: Multiple and large cysts, located in the cervical stroma, can cause significant enlargement of the cervix. CASE: A woman with two large intracervical cysts, one complex, underwent evaluation with ultrasound and magnetic resonance imaging (MRI). Hysterectomy revealed Nabothian cysts. CONCLUSION: In rare cases in which the intracervical cysts assume very large dimensions, ultrasound and MRI can aid diagnosis, but may not always prevent the need for excision or hysterectomy.

Decreased TFF2 expression in the gastric antrum in patients infected with CagA-positive Helicobacter pylori.

BACKGROUND: The trefoils factor family is a relatively new family of peptides. Their abundant expression in the epithelial cells of the gastrointestinal tract in the normal physiological state and in various ulcerative conditions suggests an important role in mucosal defense and repair. Infection with Helicobacter pylori interferes with normal mucosal activity. OBJECTIVES: To investigate whether H. pylori infection alters the expression of trefoils TFF1 and TFF2 in the gastric mucosa of patients with H. pylori-associated chronic active gastritis, positive or negative for the CagA strain. METHODS: During investigation for dyspepsia, gastric biopsies and blood samples were obtained from patients who underwent upper gastrointestinal endoscopy. Rapid urease testing, histology for determination of H. pylori-associated CAG and Western analysis for TFF1 and TFF2 expression with antisera were performed. CagA state was determined using a commercial kit. RESULTS: TFF2 expression was significantly reduced in both groups of patients with H. pylori-associated CAG compared to healthy patients without H. pylori infection, particularly in CagA-positive patients. TFF1 expression showed a tendency of reduction (not significant) in this group only. CONCLUSIONS: These results suggest that H. pylori-associated CAG has a deleterious effect on the expression of TFF2 in the gastric antrum. This reduced expression may contribute to the damage induced to the gastric mucosa by H. pylori.

EUS clarifies the natural history and ideal management of GISTs.

BACKGROUND/AIMS: Gastrointestinal stromal tumors (GISTs) are mesenchymal gastrointestinal tumors expressing C-kit (CD117). Endoscopic ultrasonography (EUS) evaluations of GISTs can help determine optimal therapy and follow-up care. The current study assesses the natural history of 100 GISTs evaluated by EUS, and the impact of EUS on their management. METHODOLOGY: Retrospective review of 2600 EUS files performed over 11 years identified 100 patients with GISTs. Relevant data from all appropriate files and interviews with patients or family of deceased patients were tabulated regarding the GISTs. RESULTS: Every GIST had definitive cytology (n=43) or histology. Seventy of the 100 patients underwent more than one evaluation. Size of the GISTs at initial diagnosis averaged 20.5 mm and at follow-up examination 23.2 mm. Fourteen of 70 GISTs showed significant enlargement (> 1 mm/month). Enlargement during follow-up of GISTs was significantly more common with GISTs over 17 mm at initial diagnosis (p<0.018). Thirty-four were excised (7 endoscopically). Clinically asymptomatic GISTs tended to be smaller. Thirteen percent of GIST patients had second primary malignancy. CONCLUSIONS: EUS elucidates GIST natural history and can help guide management. GISTs larger than 17 mm should be monitored by EUS and considered for more aggressive treatment. Searching for synchronous tumors in patients with GISTs should be considered.

Fatal thoracic aortic aneurysm and dissection in a large family with a novel MYLK gene mutation: delineation of the clinical phenotype.

BACKGROUND: Thoracic and abdominal aortic aneurysms and dissection often develop in hypertensive elderly patients. At higher risk are smokers and those who have a family history of aortic aneurysms. In most affected families, the aortic aneurysms and dissection is inherited in an autosomal dominant manner with decreased penetrance and variable expressivity. Mutations at two chromosomal loci, TAA1 at 11q23 and the TAA2 at 5q13-14, and eight genes, MYLK, MYH11, TGFBR2, TGFBR1, ACTA2, SMAD3, TGFB2, and MAT2A, have been identified as being responsible for the disease in 23% of affected families. RESULTS: Herein, we inform on the clinical, genetic and pathological characteristics of nine living and deceased members of a large consanguineous Arab family with thoracic aortic aneurysm and dissection who carry a missense mutation c.4471G > T (Ala1491Ser), in exon 27 of MYLK gene. We show a reduced kinase activity of the Ala1491Ser protein compared to wildtype protein. This mutation is expressed as aortic aneurysm and dissection in one of two distinct phenotypes. A severe fatal and early onset symptom in homozygous or mild late onset in heterozygous genotypes. CONCLUSIONS: We found that MYLK gene Ala1491Ser mutation affect the kinase activity and clinically, it presents with vascular aneurysms and dissection. We describe a distinct genotype phenotype correlation where; heterozygous patients have mild late onset and incomplete penetrance disease compared with the early onset severe and generally fatal outcome in homozygous patients.

Bone marrow fibrosis and caseating granulomas associated with intravesicular BCG treatment.

Intravesicular instillation of bacille Calmette-Guérin (BCG), an attenuated variant of the virulent strain of Mycobacterium bovis, has been extensively used as the treatment of choice for bladder cancer since first approved in 1976. Adverse effects of treatment are rare and mostly local, with systemic reactions complicating up to 5% of patients. The majority of serious side effects have occurred in immunocompromised patients. Even more rare is bone marrow involvement. Only two previous cases are reported in the medical English literature; in these cases, the granulomas were non-caseating. We describe a patient who developed fever and pancytopenia associated with mycobacterial infection after intravesicular instillation of BCG. Caseating granulomas were seen in the bone marrow biopsy and, later, bone marrow fibrosis developed gradually, in spite of successful treatment with antimycobacterial agents.

Phenotype of microsatellite unstable colorectal carcinomas: Well-differentiated and focally mucinous tumors and the absence of dirty necrosis correlate with microsatellite instability.

The phenotypic markers of colorectal carcinomas with microsatellite instability have been widely studied and include mucinous or poor differentiation, prominent host response, a circumscribed growth pattern, histologic heterogeneity, and right-sided location. As part of a population-based case-control study of colorectal cancer in northern Israel, we reviewed the pathology and microsatellite status of 528 consecutively diagnosed colorectal cancers. Phenotypic analysis was performed by one pathologist (J.K.G.) and included assessment of grade, mucinous histology (>50%, or focal), histologic heterogeneity, growth pattern, necrosis, and host response. Microsatellite status was determined on microdissected portions of formalin-fixed, paraffin-embedded tissue using a panel of 5 NCI consensus primers. Fifty-two of 528 colorectal carcinomas were microsatellite unstable (9.85%). Multivariate analysis found that >2 tumor infiltrating lymphocytes per high power field (p <0.0001), the lack of dirty necrosis (p = 0.0054), a Crohn's-like host response (p = 0.0064), right-sided location (p = 0.032), well or poor differentiation (p = 0.037), and any mucinous differentiation (p = 0.039) were independent predictors of microsatellite instability. Tumor infiltrating lymphocytes were the single best histologic predictor of microsatellite instability. The absence of dirty necrosis and the presence of well-differentiated tumors and tumors with only focal mucinous differentiation were also important markers for microsatellite instability that have not been emphasized previously. The combination of >2 tumor infiltrating lymphocytes per high power field and/or any mucinous differentiation and/or the absence of dirty necrosis identified all MSI-H tumors in this study.

Familial juvenile hypertrophy of the breast.

OBJECTIVE: Juvenile (virginal) hypertrophy of the breast (JHB) is a relatively rare condition leading to gigantomastia in peripubertal females. The pathology is limited usually to the breast, with otherwise normal growth and development and without any other deformities. The rapid growth of the breast (bilateral or unilateral) in adolescent girls leads to significant physical and psychological difficulties. This gigantomastia is treated surgically by breast reduction or mastectomy and its modification. Familial JHB was described only once in the literature, and its etiology is unknown. RESULTS: We report here on a familial pattern of juvenile hypertrophy of the breast accompanied by congenital anonychia. To the best of our knowledge, this is the first report of such a presentation. Our study dealt with four members of the same family, related through their fathers, enduring congenital anonychia of hands and feet with no functional limitation and who showed rapid uncontrolled breast enlargement in prepubertal age. This was severe enough to cause the curtailment of their social activity and cessation of schooling. The mothers of all four patients had normal breasts and nails, whereas their fathers had anonychia. The genetic basis for the association between the two clinical findings is yet to be determined. CONCLUSION: The four girls underwent breast reduction surgery.