Is flow cytometry useful in determining central nervous system involvement in patients with hematological malignancy? Analysis with a prospective cohort.

Central nervous system (CNS) involvement occurs in approximately 5-15% of patients in hematological malignancies. Early diagnosis and treatment is essential for a successful approach to CNS involvement. The gold standard method for diagnosis is cytological evaluation, but its sensitivity is low. Flow cytometry (FCM) from cerebrospinal fluid(CSF) is another method used to identify small groups of cells with abnormal phenotype. In our study, we compared FCM and cytological findings in the evaluation of CNS involvement in our patients with hematological malignancies. 90 patients [58 males, 32 females] were included in the study. CNS involvement was positive in 35(%38.9) patients, negative in 48(%53.3) patients, and suspicious (atypical) in 7(%7.8) patients by flow cytometry and it was positive in 24(%26.7) patients, negative in 63(%70) patients, and atypical in 3(%3.3) patients by cytology. While the sensitivity and specificity were found to be respectively 68.5% and 100% by cytology, it was found to be 94.2% and 85.4% by flow cytometry. Flow cytometry, cytology and MR findings were significantly correlated with each other in both prophylaxis (p < 0.001) and patients with prediagnosis of CNS involvement. Although the gold standard diagnostic method in the diagnosis of CNS involvement is cytological, its sensitivity is low and it can give false negative results at a rate of 20-60%. Flow cytometry is an ideal objective and quantitative method for identifying small groups of cells with abnormal phenotype. Flow cytometry can be used routinely in the diagnosis of CNS involvement in patients with hematological malignancies with cytology, since it can detect fewer malignant cells, has a higher sensitivity, and provides easy and faster results.

Risk of Malignancy Using the Diagnostic Categories Proposed by the World Health Organization International System for Reporting Pancreaticobiliary Cytopathology.

BACKGROUND: The World Health Organization (WHO) proposed an updated reporting system for pancreaticobiliary cytology, which moves low-grade malignancies to "positive for malignancy" group and serous cystadenoma to "negative for malignancy" group. The WHO system also created two new categories, namely, pancreatic neoplasia-low grade (PaN-Low) and pancreatic neoplasia-high grade (PaN-High), which includes neoplastic mucinous cysts and stratifies them according to their cytologic atypia. The risk of malignancy (ROM) of the new categories of the WHO system needs to be defined. METHODS: Cytologic slides of all patients, who underwent endoscopic ultrasound (EUS)-guided fine needle aspiration (FNA) biopsy at our institution from January 2010 to December 2021 and had a histopathological or clinical follow-up of at least 6 months, were reviewed and reclassified under the Papanicolaou Society of Cytopathology (PSC) and WHO reporting systems. The absolute ROM was calculated for each category of both reporting systems. RESULTS: A total of 420 EUS-FNA samples from 410 patients were reviewed and reclassified. The absolute ROM for the proposed WHO system was 35% for "nondiagnostic," 1.0% for "negative for malignancy," 69.0% for "atypical," 11% for "PaN-Low," 100% for "PaN-High," 91% for "suspicious for malignancy," and 100% for "malignant." Comparatively, the absolute ROM under the PSC reporting system was 34% for "nondiagnostic," 1.0% for negative (for malignancy), 50.0% for "atypical," 0.0% for "neoplastic: benign," 16% for "neoplastic: other," 88% for "suspicious for malignancy," and 100% for "positive or malignant." CONCLUSION: The proposed WHO international reporting system has advantages regarding risk stratification improvement and case management.

Renal Clear Cell Carcinoma with Unknown Primary: Is It Real Entity or Just Hypothesis?

Renal cell carcinoma (RCC) is a common type of cancer, and about 25% of patients are diagnosed at an advanced stage. Bone metastasis is common in RCC, but instances of bone metastases without a primary kidney tumour have rarely been reported. Here, we report a case of a 56-year male patient who presented with extensive bone metastases. The biopsy was reported as clear cell RCC and no primary renal tumour was detected by positron emission tomography and magnetic resonance imaging. Pazopanib was initiated and progression was detected in the 6th month of treatment. Nivolumab was initiated as second line therapy. In the literature, primary unrecognised RCC cases are found only as case reports. This case highlights the importance of clinicians' consideration of possible metastatic RCC in a patient with pathologically diagnosed clear cell carcinoma, even if no primary renal tumour is identified. Key Words: Renal cell carcinoma, Immunohistochemistry, Diagnosis, Kidney, Cancer.

Cross-organ sensitization between the prostate and bladder in an experimental rat model of lipopolysaccharide (LPS)-induced chronic pelvic pain syndrome.

BACKGROUND: The aim of the current study was to investigate the effects of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) on bladder function via prostate-to-bladder cross-sensitization in a rat model of lipopolysaccharide (LPS)-induced prostate inflammation. METHODS: Male rats were intraprostatically injected with LPS or saline, serving as control. Micturition parameters were examined in a metabolic cage 10 or 14 days later. Subsequently, to evaluate bladder function, cystometry was performed. Micturition cycles were induced by saline infusion and cholinergic and purinergic contractile responses were measured by intravenous injection with methacholine and ATP, respectively. Thereafter, the prostate and bladder were excised and assessed histopathologically for possible inflammatory changes. RESULTS: Metabolic cage experiments showed increased urinary frequency in rats with LPS-induced CP/CPPS. Cystometry showed a significant increase in the number of non-voiding contractions, longer voiding time and lower compliance in CP/CPPS animals compared to controls. Induction of CP/CPPS led to significantly reduced cholinergic and purinergic bladder contractile responses. Histopathological analysis demonstrated prostatic inflammation in CP/CPPS animals. There were no significant differences between the groups regarding the extent or the grade of bladder inflammation. Prostate weight was not significantly different between the groups. CONCLUSIONS: The present study shows that prostate-to-bladder cross-sensitization can be triggered by an infectious focus in the prostate, giving rise to bladder overactivity and alterations in both afferent and efferent signalling. Future studies are required to fully understand the underlying mechanisms.

Prostate-to-bladder cross-sensitization in a model of zymosan-induced chronic pelvic pain syndrome in rats.

BACKGROUND: The aim of the present study was to investigate the effects of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) on bladder function and pathophysiology. METHODS: To create a model for CPPS, rats were intraprostatically injected with zymosan or saline, serving as control. Metabolic cage experiments were performed 7, 14, or 21 days after zymosan injection and after 14 days in the control group. Thereafter, cystometry was performed in which simulated micturition cycles were induced by saline infusion and contractile responses to the cholinergic agonist methacholine and the purinergic agonist ATP were measured. Following cystometry, the prostate and urinary bladder were excised and assessed histopathologically for possible inflammatory changes. RESULTS: Metabolic cage data revealed a significantly increased urinary frequency in zymosan treated rats. Likewise, the volume per micturition was significantly lower in all CPPS groups compared to controls. Cystometry showed a significant increase in the number of nonvoiding contractions, longer voiding time, and a trend towards lower compliance in CPPS rats compared to controls. Induction of CPPS led to significantly reduced cholinergic and purinergic contractile responses. Histopathological analysis demonstrated prostatic inflammation in all CPPS groups, in particular in later stage groups. Both the extent and grade of bladder inflammation were significantly higher in CPPS groups compared to controls. CONCLUSIONS: The current findings demonstrate a potential prostate-to-bladder cross-sensitization leading to symptoms of bladder overactivity and signs of bladder inflammation. Future clinical studies are required to verify the outcomes of the current study and enable advancement of patient care.

Primary mucosa-associated lymphoid tissue (MALT) lymphoma of the lacrimal drainage system in two pediatric patients.

Primary non-Hodgkin lymphoma (NHL) of lacrimal drainage system (LDS) is quite rare in children, but it is important to expedite early diagnosis in an effort to alter possible life-threatening disease since they are usually misdiagnosed as chronic dacryocystitis. In the literature, there are few examples of tumors of LDS in children. The authors herein report two pediatric cases of extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) originating from lacrimal sac in an attempt to increase the knowledge about the clinical course of NHL of LDS. Considerable care must be taken since tumors of lacrimal drainage can mimic dacryocystitis clinically and macroscopically. Two patients both attended with painless swelling in the left lacrimal sac region and epiphora of the left eye. Orbital magnetic resonance imaging showed a tumoral lesion in the left lacrimal sac region and histopathological examination of excisional biopsy specimen demonstrated MALT lymphoma in both patients. The treatment regimen comprises lacrimal sac excision within the tumor, canalicular dacryocystorhinostomy (DCR) with bicanalicular silicone intubation (BSI) combined with chemotherapy and regional radiotherapy in one case, whereas the second case received only radiotherapy after canalicular DCR with BSI. Both of them maintained clinical remission along follow-up.

Argon laser-assisted hypotony model in the rabbit.

To investigate whether ocular hypotony formation with 360 degrees endocyclophotocoagulation is possible. Twelve male New Zealand White rabbits were used. Entire ciliary body epithelium was destructed with green laser photocoagulation after pars plana lensectomy and anterior vitrectomy in six rabbits. Endocyclophotocoagulation was not performed to the remaining six rabbits (control group). Intraocular pressure (IOP) was measured preoperatively and followed up everyday in the first week and weekly until the end of month one. All of the rabbits were sacrificed and ciliary bodies were left for gross and light microscopic examination. Mean baseline IOPs were similar in laser and non-laser group (14.8 ± 1.4 (range 12.2-17.3) vs 14.4 ± 1.4 (range 12.2-15.9), p = 0.650). Mean IOP was 6.6 ± 0.45 mmHg (range 5.9-7.1) in the laser group and 11.5 ± 1.2 mmHg (range 10.2-13.4) in the non-laser group in postoperative day 1. IOP was below 4 mmHg in all eyes on the second day and after in laser group. In the macroscopic evaluation, the entire ciliary body had a white (loss of pigmentation) and atrophic appearance in all of the eyes in the laser-treated group compared to non-laser group. In the laser group, light microscopic examination demonstrated a severe 360 degrees disruption of ciliary processes. Ciliary processes were covered with fibrin exudation consisting of fibroblasts. There was a mild inflammation with disruption or atrophy of ciliary body epithelium with cystic vacuolar degeneration. Three hundred sixty degrees endocyclophotocoagulation yielded severe ciliary epithelium damage. IOP reduction started very early and continued in hypotonic levels during follow up period.

Comparison of corneal collagen cross-linking (PACK-CXL) and voriconazole treatments in experimental fungal keratitis.

PURPOSE: To compare the antifungal efficacy of corneal collagen cross-linking with photoactivated riboflavin (PACK-CXL) and voriconazole in experimental Fusarium solani and Candida albicans keratitis models. METHODS: Sixty-four corneas of 32 New Zealand rabbits were included and divided into two main groups. Intrastromal injection of Fusarium and Candida suspensions was performed, and it was observed that keratitis was formed on the third day. Both groups were randomly separated into the following four groups: control, PACK-CXL, voriconazole and PACK-CXL combined with voriconazole. PACK-CXL was applied using 0.25% riboflavin in an accelerated Dresden protocol (total ultraviolet A dose 5.4 J/cm²). Voriconazole was applied topically as 7x1/day with a dose of 1% (10 mg/ml). Corneal buttons were excised on the tenth day, and microbiological and pathological examinations were performed. RESULTS: The PACK-CXL and PACK-CXL combined with voriconazole groups each had 100 colony-forming unit (CFU/ml) of reproduced micro-organisms compared with 500 CFU/ml in the voriconazole group and 1500 CFU/ml in the control group (p < 0.001) in the Fusarium keratitis model. The PACK-CXL combined with voriconazole group had 100 CFU/ml, the PACK-CXL group had 150 CFU/ml, and the voriconazole group had 200 CFU/ml of reproduced micro-organisms compared with 4000 CFU/ml in the control group (p < 0.002) in the Candida keratitis model. (p < 0.001). Fewer hyphae and non-specific stromal changes were observed in the pathological cross sections examined in subgroups that used CXL. CONCLUSION: There was less fungus reproduction and a lower keratitis score for Fusarium solani and Candida albicans in the treatment groups compared to the control groups, especially in groups that used PACK-CXL. These results suggest that it is useful to combine PACK-CXL treatment with medical treatment in the fungal keratitis algorithm at the early stage of the disease.

Is it easy to clinically distinguish inflammatory arthritis of bacterial origin from monoarthritis attacks of gout disease?

Acute monoarthritis is a common situation in orthopedic emergency where the patient presents with typical inflamed joint. It is hard to clinically distinguish inflammatory arthritis of bacterial origin from monoarthritis attacks of gout disease. If these two situations, which are the most common causes of acute monoarthritis, are misdiagnosed, outcomes might be catastrophic and costly. Synovial fluid analysis is the most reliable method for confirming the diagnosis although it might not always lead to definitive diagnosis. If there is clinical suspicion for crystal arthropathy, repeated examinations may provide benefits for confirming the diagnosis.

Neuroprotective effect of systemic and/or intravitreal rosuvastatin administration in rat glaucoma model.

AIM: To evaluate the neuroprotective effect of rosuvastatin, in a rat experimental glaucoma model. METHODS: Ocular hypertension was induced in right eyes of Long-Evans rats (n=30) by cauterization of three episcleral veins. Left eyes were defined as controls. Rats were divided into five groups: oral rosuvastatin, intravitreal rosuvastatin, oral+intravitreal rosuvastatin, intravitreal sham and glaucoma without intervention. Rats were sacrificed at day 14. Retinal ganglion cell (RGC) number was assessed by histopathological analysis. Terminal deoxynucleotidyl transferase-mediated dUTP-nick end-labeling (TUNEL) staining and the expression of glial fibrillary acidic protein (GFAP) in RGC layer was also examined. RESULTS: A significant intraocular pressure (IOP) elevation was seen (P=0.002). Elevated IOP resulted in a significant decrease in number of RGCs in group 5 (70.33±8.2 cells/mm(2)) when compared with controls (92.50±13.72 cells/mm(2); P=0.03). The RGC number in group 1 (92.4±7.3 cells/mm(2)) was significantly higher than group 5 (P=0.03). The numbers of RGC in groups 2, 3 (57.3±8.2 cells/mm(2), 60.5±12.9 cells/mm(2)) were comparable with that of group 5 (P=0.18 and P=0.31). The apoptosis rates with TUNEL staining were also parallel to RGC number. Animals with experimentally induced glaucoma showed an increase in retinal GFAP immunoreactivity. CONCLUSION: Decrease in RGC loss and apoptosis suggest the neuroprotective potential of oral rosuvastatin treatment in a rat model of ocular hypertension. However intravitreal rosuvastatin showed a contrary effect and further studies are required.

Ultrasound-guided thyroid nodule fine-needle biopsies--comparison of sample adequacy with different sampling techniques, different needle sizes, and with/without onsite cytological analysis.

INTRODUCTION: The aim of this study was to compare the diagnostic adequacy of thyroid samples obtained by aspiration or capillary biopsy techniques, with 22 or 27 gauge needles, and with or without on-site cytological analysis (OCA). MATERIAL AND METHODS: Four hundred patients with thyroid nodules underwent ultrasound (US)-guided fine-needle biopsies. Patients were divided into eight groups according to needle size (22 vs. 27 gauge), biopsy technique (aspiration vs. capillary), and whether or not OCA was performed. Sample adequacy rates were calculated for each group and subgroups and compared using chi-square tests. RESULTS: When all nodes were evaluated (n = 400), the adequacy rate was significantly greater with the capillary than with the aspiration technique (97% vs. 91.5%, p = 0.032) and when OCA was than was not performed (97% vs. 91.5%, p = 0.032). When only solid nodules were evaluated (n = 205) the adequacy rate was also significantly greater with the capillary than with the aspiration technique (98.9% vs. 89.7%, p = 0.008) and when OCA was than was not performed (97.9% vs. 89.6%, p = 0.014). In contrast, the adequacy rate was similar for 22 and 27 gauge needles (94.2% vs. 93.1%, p = 0.733). CONCLUSIONS: Optimal results were obtained with the capillary technique and OCA. The capillary technique and OCA should be the preferred approach in thyroid nodule biopsy, optimising adequacy rates and patient comfort.

What is the malignancy risk for atypia of undetermined significance? Three years' experience at a university hospital in Turkey.

BACKGROUND: The Bethesda System for Reporting Thyroid Cytopathology (BSRTC) provides uniform diagnostic terminology for communication between pathologists and clinicians. Each diagnostic category is associated with a specific risk of malignancy and a recommendation for its management. The indeterminate diagnostic categories of atypia of undetermined significance/follicular lesion of undetermined significance (AUS/FLUS) present a major challenge for both pathologists and clinicians. We report our institution's 3 years' experience with the AUS/FLUS category and follow-up of these patients. METHODS: A retrospective analysis was conducted for all thyroid fine-needle aspirations (FNAs) between July 2010 and July 2013. During this period, 9242 nodules from 4916 patients were reported according to the BSRTC guidelines. We adopted the AUS terminology in our practice to refer to both AUS, and FLUS. RESULTS: Of the 4916 patients, 347 (7%) were diagnosed as AUS. The malignancy risk for patients who underwent surgical resection after initial diagnosis of AUS was 22.8%, whereas that for patients who underwent a second FNA and surgical resection was 36%. When we included patients with second FNA and without surgery, the malignancy risk was 15.7%. CONCLUSIONS: The malignancy risk for AUS reported in the present study is consistent with those reported previously and is higher than those anticipated according to the Bethesda System. This supports that a multimodal approach (clinical, radiologic, and cytopathologic) is necessary for the management of thyroid nodules diagnosed as AUS. Therefore, we suggest that the recommendation for repeat FNA following an initial diagnosis of AUS should be based on a multimodal approach for each particular patient.

Resolution of inflammatory colitis with pegfilgrastim treatment in a case of severe congenital neutropenia due to glucose 6 phosphatase catalytic subunit-3 deficiency.

Glucose 6 phosphatase catalytic subunit-3 (G6PC3) deficiency is a heterogenous disorder characterized by severe congenital neutropenia and a variety of extrahematopoietic manifestations. Inflammatory bowel disease like colitis is an uncommon complication of G6PC3 deficiency, described only in adolescent and adults. Herein, we describe inflammatory colitis in a 10-year-old girl with severe congenital neutropenia due to G6PC3 deficiency while she was on a high-dose filgrastim. Switching from filgrastim to (pegylated filgrastim) Pegfilgrastim led to rapid resolution of colitis, weight gain, and decreased infections. Pegfilgrastim seems to be a better remedy for treatment of G6PC3 deficiency complicated with inflammatory bowel disease.

Near infrared spectroscopy to diagnose experimental testicular torsion: comparison with Doppler ultrasound and immunohistochemical correlation of tissue oxygenation and viability.

PURPOSE: Near infrared spectroscopy measures tissue oxygenation even when there is complete cessation of blood flow. We evaluated near infrared spectroscopy to diagnose testicular torsion and blindly compared its accuracy with that of Doppler ultrasound. We also compared it by immunohistochemical evaluation of hypoxia and cell viability. MATERIAL AND METHODS: Rats were randomized to 4 groups, including group 1-720-degree unilateral torsion, group 2-360-degree unilateral torsion, group 4-sham operation and group 4-720-degree unilateral torsion followed by surgical torsion reduction at minute 180. Near infrared spectroscopy and Doppler ultrasound were done blindly at minutes 0, 5, 30, 60, 180 and 400. All torsed and contralateral testicles were excised for pathological examination using hypoxia inducible factor-α for hypoxia and the TUNEL test for apoptosis. We compared all groups with the contralateral testis and the sham operated group. RESULTS: All blinded, near infrared spectroscopy measurements correctly identified the torsed testis after minute 5. Median oxygen saturation in groups 1 and 2 was significantly different compared to that in the sham operated group after minute 5. In group 4 near infrared spectroscopy detected detorsion with the loss of a significant oxygen saturation difference between the affected and the contralateral testicle after detorsion. At minute 400 median oxygen saturation in group 4 was not statistically different compared to that in the sham operated group (p = 0.09) but it was significantly different compared to that in groups 1 and 2 (p <0.001). In each torsed testis oxygen saturation was at least 18.75% lower than in the contralateral testis. In groups 1 and 2 hypoxia inducible factor-α staining in torsed testicles was significantly greater than that in the contralateral organ and the sham operated group. In group 4 hypoxia inducible factor-α staining after detorsion was significantly decreased compared to that in groups 1 and 2. There was no significant difference in the apoptotic index between the experimental and the contralateral testis or the sham operated group. CONCLUSIONS: Near infrared spectroscopy is as effective but quicker than Doppler ultrasound for detecting testicular torsion without a radiologist. Near infrared spectroscopy accurately reveals oxygen saturation, which is more vital than blood flow, on which Doppler ultrasound focuses.

Mechanically induced experimental knee osteoarthritis benefits from anti-inflammatory and immunomodulatory properties of simvastatin via inhibition of matrix metalloproteinase-3.

BACKGROUND: We investigated the anti-inflammatory and immunomodulatory effect of simvastatin on articular cartilage via the inhibition of matrix metalloproteinase-3 (MMP-3), a matrix-degrading enzyme, in a mechanically induced experimental osteoarthritis (OA) animal model. MATERIALS AND METHODS: Twenty-seven albino Wistar rats were divided in three groups of equal number. Unphysiologic loading of articular cartilage was simulated by transecting anterior cruciate ligaments of the right knees of 18 rats consisting of groups 1 and 2. Nine animals in group 2 received orally administered simvastatin 20 mg/kg per day by gavage for 8 weeks. Animals in group 3 were sham operated. All animals were sacrificed at postoperative 8 weeks. Effects of simvastatin on disease progression was evaluated by documenting OA changes in cartilage specimens using Osteoarthritis Research Society International (OARSI) OA cartilage histopathology assessment system scores combined with the percentage of MMP-3 expression in chondrocytes. RESULTS: Simvastatin treatment significantly down-regulated the percentage of MMP-3 expression in chondrocytes as assessed by immunohistochemistry methods. Suppression of this matrix-degrading enzyme by simvastatin also reduced OARSI scores, suggesting the potential for statins against OA progression. CONCLUSIONS: Following knee trauma, OA initiates at the molecular level in a short period of time. Irreversible structural changes in cartilage that require demanding treatment strategies led us to focus on effective measures to prevent OA. Statins have immunomodulatory and anti-inflammatory properties independent from their serum-cholesterol-lowering effects. One of these widely used drugs, simvastatin, showed beneficial effects on OA progression and extent by reducing cartilage degradation in our experimental setting. If these results are confirmed by human trials, simvastatin might be considered by orthopedic surgeons as a disease-modifying drug during the early inflammatory phase of posttraumatic OA.

Four different malignancies in one patient: a case report.

Cancer survivors have a higher risk of new primary cancer, in the same or in another organ, than the general population. We report a 78-year-old women who has metachronous quadruple adenocarcinoma, includes bilateral breast cancer, ovarian cancer and retroperitoneal neuroendocrine carcinoma. The development of second cancer in cancer survivors can be expected but third or higher order malignancies are rare.

Matrix metalloproteinase-9 expression in retinal ganglion cell layer and effect of topically applied brimonidine tartrate 0.2% therapy on this expression in an endothelin-1-induced optic nerve ischemia model.

The purpose of this research is to investigate the expression of matrix metalloproteinase-9 (MMP-9) in retinal ganglion cells (RGC) and the impact of topically applied brimonidine tartrate 0.2% (BMD) on this expression in an endothelin-1 (ET-1)-induced chronic optic nerve (ON) ischemia model of rabbit. Osmotically driven minipumps were implanted in one eye of 16 New Zealand albino rabbits to deliver ET-1 at the constant rate of 0.5 microl/h for 2 weeks. ET-1 was given with (group 3) and without topical BMD therapy (group 1). Groups 2 and 4 were taken as controls. MMP-9 expression by immunohistochemically and proportion of cells undergoing apoptosis in RGC layer were investigated. The correlation between the MMP-9 immunopositivity and the proportion of cells undergoing apoptosis in the RGC layer was evaluated. MMP-9 immunopositivity was found to be significantly higher in both groups 1 and 3 compared to that of the controls. There was no difference between groups 1 and 3 regarding MMP-9 expression (p = 0.495). A positive correlation was found between the proportion of cells undergoing apoptosis and MMP-9 expressions in the RGC layer in group 1 (p = 0.031, r = 0.754). MMP-9 expression in the RGC layer seems to significantly increase in the ET-1-induced chronic ON ischemia model. Topical BMD therapy does not seem to affect this MMP-9 expression.