Pitfalls of cytological diagnosis of tumours of the pancreaticobiliary tract.

The recent introduction of the WHO cytology classification of pancreatobiliary tumours aimed to improve the diagnosis and management of these tumours. The present paper briefly describes the methods of diagnosis. Emphasis is then put on a detailed comparison of the previous Papanicolaou classification and the new WHO classification and description of the changes brought about by the introduction of the WHO classification. In the last part of the paper, we present interesting cases from our practice illustrating possible diagnostic pitfalls of cytological evaluation.

Case report: Gaucher disease in trepanobiopsy of 16yo woman examined for suspected myelodysplastic syndrome.

Gaucher disease is an autosomal recessive disease belonging to the so-called storage diseases. More than 300 mutations of the GBA1 gene encoding the β-glucocerebrosidase enzyme are known. It is a very rare disease in the Czech Republic. Currently 35 patients are treated. In our case report, we present the case of a 16 year old female patient attending the Clinic of Pediatric Medicine at the University Hospital in Ostrava. Since 2007, the patient has suffered prolonged thrombocytopenia, at the time with progression, and splenomegaly, which has not been further investigated. Trepanobiopsy was sent to the Department of Pathology with suspicion of myelodysplastic syndrome in May of 2018. In the biopsy examination, the individual bloodline did not show dysplastic features and the number of blasts was not increased. The marrow interstitium was 70% permeated with gaucher cells with intraplasmatic fibrous material. Cells were in the appearance of „crumpled paper“ and expressed CD68 in immunohistochemical stain and in histochemical examination of PAS and iron (Fe) staining. Based on a morphological finding, Gauchers disease was suspected. Repeated bone marrow aspirates were subsequently captured by gaucher cells, and a next biochemical examination showed a β-glucocerebrosidase enzyme decrease of activity. Gaucher disease is a progressive disease that requires early diagnosis with the onset of therapy.

The impact of standard protocol implementation on the quality of colorectal cancer pathology reporting.

BACKGROUND: The aim of the study is to assess the influence of standardized protocol implementation on the quality of colorectal cancer histopathology reporting. METHODS: A standardized protocol was created based on the recommendations of The College of American Pathologists. The impact of this protocol was measured by comparing frequencies of assessed parameters in histopathology reports before and after implementation. RESULTS: In total, 177 histopathology reports were included in this study. The numbers of harvested lymph nodes were 12.4 ± 5.2 (colon) and 12.6 ± 5.4 (rectum) before protocol; and 17.1 ± 6.5 (colon), and 16.6 ± 7.0 after protocol implementation; differences were statistically significant. The recommended minimum of 12 lymph nodes was not achieved in 42.8 % (colon) and 45.7 % (rectum) of specimens before, and in 10.4 % (colon) and 17.7 % (rectum) of specimens after protocol implementation; differences were statistically significant. There were no differences in histopathology assessment of proximal and distal resection margins, grading assessment, TNM staging recording, and number of positive findings of microscopic tumor aggressiveness. The findings of tumor budding, tumor satellites, and assessment of microscopic tumor aggressiveness were more frequent after protocol implementation. Histopathology reports of rectal specimens contained assessments of the macroscopic quality of mesorectum, circumferential resection margin, and neoadjuvant therapy effect (if administered) only after protocol introduction. CONCLUSIONS: A standardized protocol is a valuable and effective tool for improving the quality of histopathology reporting. Its implementation is associated with more precise specimen evaluation, higher numbers of harvested lymph nodes, and improved completeness of histopathology reports.

Analysis of Histopathological Findings of Lung Carcinoma in Czech Black Coal Miners in Association with Coal Workers' Pneumoconiosis.

Coal miners with coal workers' pneumoconiosis (CWP, J60 according to ICD-10) were previously found to have a significantly higher risk of lung carcinoma compared to the general male population. The presented study aimed to analyze the (i) incidence of lung carcinoma in miners, (ii) histopathological findings in cohorts with and without CWP, and (iii) effect of smoking cessation on the histopathological profile. Analyzed cohorts consisted of miners with (n = 3476) and without (n = 6687) CWP. Data on personal and working history obtained from the medical records were combined with information on lung cancer from the Czech Oncological Register and histopathological findings. Statistical analysis was performed using non-parametric tests and the incidence risk ratio at the significance level of 5%. In 1992-2015, 180 miners (2.7%) without CWP and 169 (4.9%) with CWP, respectively, were diagnosed with lung carcinoma. The risk of lung cancer in miners with CWP was 1.82 (95% CI: 1.48-2.25) times higher than in those without CWP. Squamous cell carcinoma (37%) was the most common histopathological type, followed by adenocarcinoma (22%) and small cell carcinoma (21%). A statistically significant difference between the cohorts (p = 0.003) was found in the histopathological subtypes, with the incidence of small cell carcinoma being 2 times higher in miners without CWP than in those with CWP. Only a few individuals with lung carcinoma were non-smokers. The incidence of small cell carcinoma, which is strongly associated with smoking, is significantly higher in miners without CWP. Smoking constitutes the most important risk factor for developing lung carcinoma even in that cohort. However, CWP remains a very important risk factor.

Pseudoactinomycotic radiate granules in the maxillary sinus. A case report.

BACKGROUND: Pseudoactinomycotic radiate granules are biologically inactive formations simulating true actinomycotic granules. They occur mainly in the female genital tract. Here we describe a previously unreported case of these granules in a maxillary sinus. METHODS AND RESULTS: A 74-year old man underwent maxillectomy for a carcinoma of the right alveolar ridge. In the tumor-free sinus mucosa, a deposit of condensed mucus, cell debris and eosinophilic granules, surrounded by leukocytes, were seen. These Gram-positive granules were refractile, not bi-refringent and revealed radiate pattern with peripheral club-like thickening. Alcian blue staining was negative, PAS reaction was present at the periphery. Ziehl-Neelsen, Grocott and Kongo red reactions tested negatively. In the mucoid surroundings, strong IgM, moderate IgG, weak IgA and IgG4, but no IgD immunoreactivity was present. CONCLUSIONS: PAMRAGs in the maxillary sinus have not been described so far. Despite their extremely rare occurence, they must be taken into consideration in histopathological differential diagnosis of actinomycosis of that anatomic location.

Novel approach to computerized breath detection in lung function diagnostics.

BACKGROUND: Breath detection, i.e. its precise delineation in time is a crucial step in lung function data analysis as obtaining any clinically relevant index is based on the proper localization of breath ends. Current threshold or smoothing algorithms suffer from severe inaccuracy in cases of suboptimal data quality. Especially in infants, the precise analysis is of utmost importance. The key objective of our work is to design an algorithm for accurate breath detection in severely distorted data. METHODS: Flow and gas concentration data from multiple breath washout test were the input information. Based on universal physiological characteristics of the respiratory tract we designed an algorithm for breath detection. Its accuracy was tested on severely distorted data from 19 patients with different types of breathing disorders. Its performance was compared to the performance of currently used algorithms and to the breath counts estimated by human experts. RESULTS: The novel algorithm outperformed the threshold algorithms with respect to their accuracy and had similar performance to human experts. It proved to be a highly robust and efficient approach in severely distorted data. This was demonstrated on patients with different pulmonary disorders. CONCLUSION: Our newly proposed algorithm is highly robust and universal. It works accurately even on severely distorted data, where the other tested algorithms failed. It does not require any pre-set thresholds or other patient-specific inputs. Consequently, it may be used with a broad spectrum of patients. It has the potential to replace current approaches to the breath detection in pulmonary function diagnostics.

Dapsone treatment is efficient against persistent cutaneous and gastrointestinal symptoms in children with Henoch-Schönlein purpura.

BACKGROUND: Henoch-Schönlein purpura (HSP) is a systemic disorder characterized by leukocytoclastic vasculitis involving the capillaries and by the deposition of IgA immune complexes. An association between HSP and atypical bacteria is uncommon in children. METHODS AND RESULTS: Here we report three cases of children, aged 5, 4 and 16 years, who were diagnosed with HSP associated with Mycoplasma pneumoniae or Chlamydia pneumoniae infection. In all presented cases, persistent cutaneous manifestations and abdominal pain were resistant to antibiotics and corticosteroids, but resolved during 48 h after the introduction of dapsone. No adverse effects of treatment were observed. CONCLUSION: Dapsone, a sulphone with an anti-inflammatory activity, showed remarkable therapeutic efficacy against rash and gastrointestinal symptoms in children with HSP. Its administration should be considered particularly in persistent cutaneous form of HSP.

Expression and significance of survivin in colorectal high grade and low grade adenomas.

We examined immunohistochemically the expression pattern of a potential tumor biomarker survivin in a panel of 116 tubular adenomatous polyps to determine its association with clinicomorphological parameters such as age of patients, size of polyps, degree of dysplasia and polyp localization. In each section, the subcellular localization of survivin antigen and the intensity of staining were assessed. Overall, survivin was expressed in 90 cases (77.6%). Cytoplasmic positivity was observed in 46/116 cases (39.7%), while nuclear and combined nuclear and cytoplasmic reaction in 44/116 cases (37.9%). High grade dysplasia was diagnosed in 52 cases (44.8%) and low grade dysplasia in 64 cases (55.2%). Statistical analysis revealed a significant correlation between subcellular survivin localization and the degree of dysplasia, size of polyps and colon localization. On the other hand, survivin expression pattern did not correlate with the age of patients. Statistically significant trend was confirmed between intensity of survivin immunoreaction and tumor size and dysplasia grade, and also the trend between negative/strong survivin intensity and polyp localization. Another statistically significant association was found between the degree of dysplasia and the size of polyps. Our findings revealed that survivin is frequently expressed in different subcellular compartments of adenoma cells. Our recent results suggest that the nuclear and combined nuclear and cytoplasmic survivin localizations are strongly associated with poor prognostic parameters in the assessment of colon adenomas. Thus, survivin may represent a promising biomarker in immunohistochemical evaluation of these lesions.

Contribution to differential diagnostics of granulomatous lesions in lung tissue.

Etiopathogenesis of pulmonary granulomatous lesions can be very varied. Pathologists find pulmonary granulomatous lesions in autopsies frequently. Diagnostics of pulmonary granulomatous lesions is based on anamnesis data, clinical examinations and morphological changes. In some cases, especially in long-term ill patients, morphological finding remains the only way to explain the case. We provide a description of an incidental autoptic finding of chronic granulomatous lesion in lung tissue in a man aged 81. Histopathological examination proved aspired granules of maize starch. Aspired granules triggered lesion of granulomatous character. At microscopic examination vegetable particles were defined to be maize starch. Granules of maize starch, which are a part of artificial nutrition labelled Caglusal, may have caused granulomatous lesion in our case.

Relationship of mismatch repair proteins and survivin in colon polyps and carcinomas.

Mismatch repair genes (MMR) play an essential role in DNA repair. MMR mutations predominantly in MLH1, MSH2, MSH6, PMS2, and rarely in PMS1, may cause the production of abnormally short or inactivated proteins. The antiapoptotic protein survivin functions in the inhibition of apoptosis, regulation of cell division and also enhances angiogenesis. Both MMRP and survivin are considered to be powerful prognostic parameters. This study was designed to determine the relationship between MMRP and survivin in colon lesions. The study included 113 cases of colon carcinoma and 51 cases of colon polyps. Survivin expression and MMRP status were assessed by immunohistochemistry. In each section, expression, intensity of immunostaining and percentage of labeled cells were analyzed. In carcinomas, immunoreaction was detected in 100/113 cases for MLH1 (88.5%), 112/113 cases for MSH2 (99.1%), 110/113 cases for MSH6 (97.3%), and 103/113 cases for PMS2 (91.2%). Survivin was shown in 47/113 cases (41.6%). The statistical analysis confirmed a significant correlation between the expression of MMRP and survivin in the assessed parameters. All 51 polyp samples were positive for MLH1, MSH2, MSH6 and PMS2. Only 8 of those (15.7%) were positive for survivin. Statistically significant differences were observed between the expression of MMRP and survivin. In conclusion, this study revealed that MMRP may suppress the antiapoptotic function of survivin through p53 inactivation of its promoter in grade 1 and grade 2 colon carcinomas.

Carbonic anhydrase IX: a promising diagnostic and prognostic biomarker in breast carcinoma.

We examined the expression of carbonic anhydrase IX (CA IX) by immunohistochemical staining using monoclonal antibody M75 (Institute of Virology, Slovak Academy of Sciences, Bratislava) in a group of 38 fibroadenomas and 55 carcinomas of the breast. In each case, the intensity of staining, percentage of labeled cells and subcellular localization of CA IX were assessed. CA IX was detected in 11/38 fibroadenomas (28.9%). Weak cytoplasmic positivity was dominant in these positive cases. Immunohistochemical analysis of 55 carcinomas showed CA IX expression in 34 cases (61.8%). Membrane staining alone was observed in 27/55 carcinomas (49.1%), while cytoplasmic positivity was found in 4/55 cases (7.3%). Combined membrane and cytoplasmic immunostaining of CA IX was detected in 3/55 carcinomas (5.4%). The intensity of immunoreactivity varied from weak to strong. Under 50% of reactive cells were found in 9/38 fibroadenomas (23.6%) and in 29/55 carcinomas (52.7%). More than 50% of reactive cells were found in 2/38 fibroadenomas (5.3%) and in 5/55 carcinomas (9.1%). Statistical analysis confirmed significant differences in the subcellular localization, intensity of immunoreactivity and percentage of labeled cells in fibroadenomas and carcinomas (p<0.05). Our results confirmed the hypothesis that expression of CA IX may represent a valuable tumor biomarker as well as a promising diagnostic and prognostic parameter in breast cancer.

Endoscopic vs open saphenous vein harvest for coronary artery bypass grafting: a leg-related morbidity and histological comparison.

AIMS: The aim of this study was to compare coronary artery bypass grafting (CABG) using either endoscopic (EVH) or open harvest (OVH). Leg-related morbidity and histological comparison of the veins were the outcome measures. METHODS: One hundred consecutive patients scheduled for isolated CABG were randomly divided into two goups: an EVH and OVH group. Perioperative data were recorded. Patients were examined 7 days and 1 month postoperatively for leg-related morbidity. Samples for histological examination were taken from each harvested vein during the surgery. RESULTS: Postoperative pain was statistically significantly lower in the EVH group 7 days postoperatively but the incidence of haematoma was non statistically higher in the EVH group while swelling was higher in the OVH group. Almost 40% of all histological samples were described as showing endothelial damage. There was significantly more endothelial damage in the EVH than the OVH group. CONCLUSIONS: We confirmed the advantage of EVH in terms of leg-related morbidity as well as cosmetic effect. This method however, was associated with more acute endothelial damage of the graft. These results support concerns that endoscopic vein harvest may be connected with detrimental effects on vein endothelium which could promote a thrombogenic environment leading to a decrease in graft patency. This could be extremely important. The results suggest that further investigation of the long-term patency of vein grafts harvested endoscopically is required.

Survivin expression in breast lobular carcinoma: correlations with normal breast tissue and clinicomorphological parameters.

The antiapoptotic protein survivin is rarely expressed in normal adult differentiated tissues, but it is often detected in their malignant counterparts. Immunohistochemically, we evaluated survivin expression in 19 cases of normal breast tissue and 64 cases of lobular breast carcinoma. The intensity of staining, percentage of labeled cells and subcellular location of survivin were assessed. We analyzed the quantitative differences of survivin expression between normal breast tissue and carcinomas. We also correlated survivin expression pattern in carcinomas with clinicomorphological parameters such as age of patients, grade, stage and size of primary tumor, lymph node metastasis, vascular invasion as well as estrogen and progesterone status. Survivin was detected in 10/19 cases of normal breast tissue (52.6%) and in 55/64 cases of lobular breast carcinoma (86%). The statistical analysis confirmed significant correlations between the assessed parameters in normal breast and lobular carcinoma. Furthermore, the expression of estrogen correlated significantly with the subcellular localization and intensity of survivin in carcinoma. However, no significant correlation was shown with regard to other clinicomorphological parameters. Our results suggest that survivin may be a valuable diagnostic marker, as well as a new independent prognostic parameter, in lobular breast carcinoma. Finally, our data support the hypothesis that lobular and ductal breast carcinomas seem to be different clinicomorphological entities.

Plasma levels of vascular endothelial growth factor during and after radiotherapy in combination with celecoxib in patients with advanced head and neck cancer.

Celebrex and radiotherapy in advanced head and neck cancer. This phase I dose-escalation study seeks to determine the phase II recommended dose of cyclooxygenase type 2 (COX-2) inhibitor in patients with locally advanced squamous cell head and neck (H&N) cancer, treated with accelerated radiotherapy. Anti-vasculogenic effect of this treatment on serum vascular endothelial growth factor (VEGF) is examined. Patients were irradiated with curative intent (72Gy in 6weeks). Celecoxib was administered throughout the radiotherapy course. Serum VEGF level were tested during radiotherapy and in follow-up. Tumor specimens were stained to quantify the COX-2 expression. Thirty-two patients completed the treatment. The dose of celecoxib was escalated (200, 400 and 800mg bid, then de-escalated to 600mg bid). The acute toxicity related to the treatment in the first and second cohort did not reach grade III; in the third cohort three patients had grade III radiation toxicity and one had celecoxib-related toxicity. In the last fourth cohort the toxicity was acceptable. Significant VEGF level drop (p=0.011) was found between radiation day 1 and post-treatment visit. Significant decrease (p=0.022) of the VEGF level was shown in patients with high COX-2 expression in the tumor. Phase II recommended dose of celecoxib combined with accelerated radiotherapy in advanced H&N cancer was 600mg bid. A significant decrease of the post-treatment serum VEGF level compared to the initial level was noticed only in patients with high COX-2 expression in tumors.