A case of Covid-19-associated invasive pulmonary mucormycosis in a pediatric patient with a newly diagnosed diabetes.

Mucormycosis is a rare, potentially life-threatening disease that is growing during the Covid-19 pandemic. This study reports a case of an 11-year-old patient with fatal Covid-19-related pulmonary mucormicosis and diabetes mellitus with ketoacidosis. The diagnosis was set post mortem. It was based on histochemical detection of the causative agent. Massive hemoptysis due to erosion of a large pulmonary vessel caused mechanical asphyxia and lethal outcome. Pulmonary mucormycosis may be highly suspected in patients with long-term Covid-19, poorly controlled diabetes mellitus with ketoacidosis, and corticosteroid therapy. Early diagnosis and treatment with Amphotericin B are potentially curative options for this invasive fungal infection and can led to better outcome.

[MICROSATELLITE INSTABILITY AND SURVIVAL IN PATIENTS WITH ENDOMETRIAL CANCER.]

Microsatellite-unstable genotype associated with deficiency mismach repaired enzymes leads to the accumulation of a series of mutations in the coding and regulatory regions of the genes having a role in carcinogenesis in endometrial cancer (ECs). The aim of this study was to investigate the correlation between the expression of four main mismach repaired enzyme (MLH-1, MSH-2, MSH-6 and PMS-2) with patients survival. ECs from 70 women (median of 63 years, range of 43-79 years) were assessed immunohistochemically for microsatellite instability (MSI). In our study we found that the presence of MSI, determined by the absence of immunohistochemically expression of at least one of the test four of the enzyme mismach repaired system is associated, not statistically significant, with longer survival (p = 0.558). In conclusion we may state that the immunohistochemical analysis who indicates MSI in biopsy tissue is a one step forward for the determination of survival and progression of endometrial carcinoma.

[Molecular based targets and endometrial cancer].

In recent years, increasing attention has been paid to the rate of spread of endometrial carcinoma, especially in the postmenopausal period. Along with routine diagnostic methods, giving information on the location and progression of the disease, there are some morphological methods determining very accurately the correlations in the development of this type of cancer and his prognosis. Moreover--in recent years, the accumulated information about the molecular profile of this type of cancer made it possible to implement a number of new drugs against the so-called molecular therapy -'targets' in the neoplastic process. Significant proportion of cases show response rates, it is more hope in the development of more successful formulas and target -based therapy. In this review, we present and discuss the role of certain molecular markers as potential indicators of prognosis and development, as well as determining the target treatment of endometrial carcinoma.

[MICROSATELLITE INSTABILITY IN ENDOMETRIAL CANCER].

Microsatellite instability (MSI) is involved in the pathogenesis of about 30% of endometrial cancer cases. In accumulating data from the past few years MSI is shown to have both clinical and prognostic value. In the present work we are discussing the components and molecular mechanisms of functioning of the mismatch repair system (MMR). MSI is a hallmark of endometrial tumors with DNA MMR deficiency. We summarized the main pathways for accumulating mutations in coding and non-coding DNA sequences, which is a result of errors during replication of microsatellite tandem repeats. A few studies rising valuable conclusions about the correlation between MSI and endometrial cancer are revised.

Quantitative morphology in canine cutaneous soft tissue sarcomas.

Stained cytological specimens from 24 dogs with spontaneous soft tissue sarcomas [fibrosarcoma (n = 8), liposarcoma (n = 8) and haemangiopericytoma (n = 8)], and 24 dogs with reactive connective tissue lesions [granulation tissue (n = 12) and dermal fibrosis (n = 12)] were analysed by computer-assisted nuclear morphometry. The studied morphometric parameters were: mean nuclear area (MNA; µm(2)), mean nuclear perimeter (MNP; µm), mean nuclear diameter (MND mean; µm), minimum nuclear diameter (Dmin; µm) and maximum nuclear diameter (Dmax; µm). The study aimed to evaluate (1) possibility for quantitative differentiation of soft tissue sarcomas from reactive connective tissue lesions and (2) by using cytomorphometry, to differentiate the various histopathological soft tissue sarcomas subtypes in dogs. The mean values of all nuclear cytomorphometric parameters (except for Dmax) were statistically significantly higher in reactive connective tissue processes than in soft tissue sarcomas. At the same time, however, there were no considerable differences among the different sarcoma subtypes. The results demonstrated that the quantitative differentiation of reactive connective tissue processes from soft tissue sarcomas in dogs is possible, but the same was not true for the different canine soft tissue sarcoma subtypes. Further investigations on this topic are necessary for thorough explication of the role of quantitative morphology in the diagnostics of mesenchymal neoplasms and tumour-like fibrous lesions in dogs.

The "mystery" of cutaneous sarcoidosis: facts and controversies.

The reason why the cutaneous form of sarcoidosis is well known in the literature is because of its spectrum of manifestations granting it the fame of a Great Imitator. The mystery shrouding the pathogenesis of this rare cutaneous disease is still there (in spite of the fundamental progress of the various diagnostic methods in current day medicine). The production of the morphological substrate - the epithelioid cell granuloma - which is considered to be characteristic of skin sarcoidosis, could, however, also be the end result of a reaction to i) various specific infectious agents such as Leishmaniasis cutis, coccidioidomycosis, etc., ii) certain residual bacterial or other mycobacterial antigens which, at the moment of setting the diagnosis are - by definition - non-infectious but still immunogenic, as well as iii) different tumor antigens in lesional tissue or other location. Often, differentiating between sarcodiosis and a sarcoid-like reaction, based on the updated criteria for cutaneous sarcoidosis, is problematic to downright impossible. A future characterization of the genetic signature of the two conditions, as well as the implementation of additional mandatory panels for i) the identification of certain infectious or ii) non-infectious but immunogenic and iii) tumor antigens in the epithelioid cell granuloma (or in another location in the organism), could be a considerable contribution to the process of differentiating between the two above-mentioned conditions. This will create conditions for greater accuracy when setting the subsequent therapeutic approaches.

Macrophages and dendritic cells in the development of liver injury leading to liver failure.

Liver failure (LF) continues to be a serious problem due to different underlying disorders. Not only hepatocytes but Kupffer cells (KCs) and dendritic cells (DCs) are of importance in this instance. We wanted to investigate the possible role of KCs and liver DCs in the development of liver injury in patients with liver failure. Liver specimens from 23 patients who died after liver failure were examined for the presence and distribution of CD68-positive KCs and CD83-positive DCs by immunohistochemistry. The distribution of the CD83-positive DC in the sinusoidal and the periportal spaces was not even. While 39.1% of patients had a high sinusoidal density of CD83-positive cells, 60.9% demonstrated a high density of CD83-positive cells in the periportal tract. The number of CD83-positive DCs in periportal tracts in patients with advanced liver fibrosis (n=5) were high, while those with mild liver fibrosis (n=18) had low numbers of mature dendritic cells (χ2=4.107; p=0.043). In addition, all patients with intensive fibrosis had low counts of CD68-positive KC’s in portal tracts vs patients with mild fibrosis of which 67% had high counts (χ2=6.97; p=0.008). In seven of the patients with moderate steatosis (87.5%) low numbers of CD68-positive KCs were found in sinusoids, in contrast to those with severe steatosis, where 12 patients (80%) had high KC counts (χ2=13.4; p less than 0.001). The distribution and number of CD68-positive KC and CD83-positive DC reflect the progression of liver fibrosis leading to liver failure.

Angiomyolipoma of the helix without signs of systemic involvement: successful surgical approach.

Cutaneous angiomyolipoma (AMP) of the ear is an extremely rare benign mesenchymal tumour. Angiomyolipomas are the most common benign tumour of the kidney and could be associated in 20% of the cases with the tuberous sclerosis complex (TBC), in which condition, most of the patients have several angiomyolipomas affecting both of the kidneys. We report the rare case of a 66-year-old female who had an asymptomatic, solitary, nodule on the helix of the right ear for several years, which was histologically assessed as angiomyolipoma. There were no clinical signs of the tuberous sclerosis complex (TBC) or renal AML. A surgical excision was performed with an excellent therapeutic result. In contrast to renal AMLs or PEComas, which are often invasive and may involve regional nodes, cutaneous AMLs are solitary, non-invasive, and not associated with tuberous sclerosis, and are curable by simple elliptical excision.

Microsatellite instability and promoter hypermethylation of MLH1 and MSH2 in patients with sporadic colorectal cancer.

PURPOSE: Inactivation of the genes involved in DNA mismatch repair (MMR) is associated with microsatellite instability (MSI) and loss of heterozygosity (LOH). The aim of the current study was to assess the presence of MSI and promoter hypermethylation of MLH1 and MSH2 in Bulgarian PATIENTS WITH SPORADIC COLORECTAL CANCER (CRC) AND TO ANALYZE THEIR POSSIBLE EFFECT ON THE DEVELOPMENT, PROGRESSION AND PROGNOSIS OF THE DISEASE. METHODS: We examined MSI in 126 patients with sporadic CRC and the methylation status of the MLH1 and MSH2 promoter regions in the cases with MSI/LOH by using a panel of 5 microsatellite markers (BAT26, D5S346, D18S35, D2S123 and FGA) and methyl-specific PCR (MSP) of bisulfite converted DNA. RESULTS: MSI/LOH was found in 36 (28.6%) patients. Among them, 30 were analyzed for promoter hypermethylation of MLH1 and we detected hypermethylation in 15 (50%) of them, whereas promoter hypermethylation of MSH2 was observed only in one case. The presence of MSI/LOH was associated with younger age (p=0.002), more advanced stage (III/IV stage) (p=0.029), lower degree of differentiation (p=0.001), and right-sided tumor localization (p=0.0002), but not with overall survival (log rank, p=0.566). CONCLUSION: Our data suggest that sporadic CRCs with MSI/LOH are more aggressive, develop earlier and progress faster to more advanced stage. The most frequent cause of failure of DNA MMR system appeared to be the hypermethylation of CpG islands of the promoter region of MLH1, whereas the methylation of MSH2 was a rare event.

Immunohistochemical study of 3 beta-hydroxysteroid dehydrogenase in dog's perianal sinus.

In this study, the localization of 3 beta-hydroxysteroid dehydrogenase (3beta-HSD) activity in the wall of canine perianal sinus (PS) was determined. The 3 beta-HSD activity was found out both in the cytoplasm of cells, situated in the propria and forming clusters adjacently to apocrine glands and in the cytoplasm of some epithelial cells in apocrine cells' glands. The results obtained about the 3 beta-HSD activity allowed us to propose a role of this enzyme in PS development and possibly, in tumourogenesis.

Recruitment of dendritic cells in human liver with metastases.

Dendritic cells (DCs) play a key role in the generation of antitumor immune responses as the most potent professional antigen-presenting cells. In this study we examined the distribution of DCs subsets in selected areas of liver metastases and adjacent liver tissue of 74 patients with gastrointestinal cancers (14 gastric, 47 colon, and 13 rectal) using immunohistochemistry for the DCs markers S-100 protein, HLA-DR, CD1a, and CD83. S-100 protein-positive DCs were localized mainly in clusters in metastases and at the tumor border with the surrounding liver tissue, while HLA-DR-positive DCs were significantly more in number (P<0.0001) and were diffusely distributed in metastasis stroma and at the tumor border. S-100 protein-positive DCs with mature phenotype were presented around metastases and in the sinusoidal lumena, whereas S-100 protein-positive DCs with less mature phenotype based on their ultrastructure were scattered in the tumor stroma. CD1a- and CD83-positive DCs were observed predominantly in small groups or as single cells in the tumor stroma and in the invasive margin. The numbers of CD1a-positive DCs (immature) and CD83-positive DCs (mature) were comparable, but significantly lower than that of S-100 protein-positive (P<0.0001) and HLA-DR-positive cells (P<0.0001).We observed more S-100 protein-positive DCs and HLA-DR-expressing cells in the sinusoids and portal tracts of the liver tissue, surrounding metastases, than in control liver tissue. In conclusion, this study provides additional information on the functional subtypes and distribution of DCs infiltrating metastatic tissue and local liver environment in patients with liver metastases from gastrointestinal cancers.

Microsatellite instability in Bulgarian patients with endometrial cancer.

PURPOSE: Microsatellite instability (MSI) is a frequent event in different types of cancer. In several studies MSI was shown to have both clinical and prognostic value. The aim of our study was to determine the frequency of MSI in Bulgarian patients with endometrial cancer (EC) and the possible relation of this phenomenon to their clinicopathological characteristics. PATIENTS AND METHODS: A total of 33 histologically confirmed EC patients were analyzed for tumor MSI using a panel of 6 microsatellite markers. RESULTS: We identified MSI in 30% of endometrial cancer cases. Six of them had high degree of MSI (MSI-H), and 4 displayed low degree of MSI (MSI-L). CONCLUSION: The frequency of MSI in Bulgarian EC patients does not differ significantly from that reported in other European studies.

[Cytokeratin-19 in diagnosis of bone marrow metastases in patients with breast cancer - clinical results].

A lot of information about the structure and the clinical significance of citokeratins in diagnosis of malignances has been found in literature during last years. Some questions about their significance in tumor genesis and in prognosis when tumor cells are found far from the primary tumor are cleared. Our aim was to investigate bone marrow samples from 79 women with breast cancer and to detect the presence or the absence of citokeratin positive tumor cells and their significance for the prognosis of the disease. We found that citokeratin 19 provides a good estimation for presence of tumor cells in bone marrow of patients with breast cancer. Citokeratin positive samples are a marker for worse prognosis in patients with breast cancer than citokeratin 19 negative ones.

Significance of tenascin-C, fibronectin, laminin, collagen IV, alpha5beta1 and alpha9beta1 integrins and fibrotic capsule formation around liver metastases originating from cancers of the digestive tract.

The formation of a fibrotic capsule around liver metastases may functionally act as a barrier to local invasion. However, the prognostic significance of exstracellular matrix (ECM) and of some integrins' deposition around liver metastases remains unclear. An immunohistochemical investigation was carried out on 55 patients with synchronous liver metastases from colorectal and gastric cancers. Encapsulated metastases were detected in 60% of the cases. The 'non-capsular' cases showed clear immunostaining for tenascin-C, fibronectin, collagen IV, laminin, alphaSMA and integrins. On opposite, most of the cases with 'capsule' were negative for the studied ECM proteins and the two integrins. The patients with 'capsular' pattern had significantly longer median survival after the surgery compared to those with non-encapsulated metastases. The presence of tenascin, fibronectin, fibronectin receptor and laminin, as well as the strong immune signal for alphaSMA and collagen type IV in the sinusoids attached to the liver metastases was associated with a worse prognosis. The cells, forming ECM in the sinusoids attached to metastases in the 'non-capsular' pattern were alphaSMA-positive myofibroblasts. It was shown ultrastructurally that they were HSCs. The results indicate that fibrotic capsule formation is associated with longer survival after surgery. The appearance of tenascin-C and of its receptor at the periphery of liver metastases could be used as a sign of invasiveness.

[Expression of human papilloma virus (HPV) and P16ink4a in cervical intraepithelial neoplasia (CIN) and papilloma].

UNLABELLED: p16ink4a Is a tumor-supressor gene and a key regulator of the cell cycle. The neoplastic cervical epithelium (squamous and cylindric) intensely expresses the cyclin-dependent kinase inhibitor p16ink4a. It was shown that the expression of p16ink4a in the squamous cervical cancer was induced by HPV. The aim of our study was to reveal the frequency of expression of p16ink4a and of HPV in the different grades of CIN and in the papilomatous cervical lesions. We used biopsy material (n=11) and cone biopsies (n=10), collected from women with different grades of CIN and with papillomatous lesions, having different grades of koilocytotic dysplasia. The biopsy material, collected from one woman with squamous cervical cancer was also studied. Using the immunohistochemical procedure we determined the expression of p16ink4a and HPV in the changed squamous cervical epithelium. RESULTS: We found that the neoplastic epithelium in CIN II and CIN III intensely expressed p16ink4a. We observed positive immune reaction for HPV in only 22.7% from the cases with papillomatous lesions, having koilocytic dysplasia. In conclusion we can state that p16ink4a could be used as a marker for the exact determination of CIN lesions in the cervix uteri. The koilocytic dysplasia, determined histologically was not always HPV-positive.

Morphological and immunocytochemical investigations on mast cells in porcine ureter.

Morphological, morphometric, histochemical and immunocytochemical investigations on mast cells, located in the wall of ureter of 8 months aged pigs were performed. Mast cells were found in all three layers of ureteral wall, but their distribution was irregular and the number unequal. It was established that alcian blue (AB)-positive mast cells were significantly more than toluidine blue (TB)-positive mast cells. A statistically significant smaller number of both AB and TB-stained mast cells were observed in the tunica mucosa. The largest number of mast cells was found in the tunica muscularis. In the adventitia, mast cells were higher in number in the main connective tissue than in the connective tissue near the blood vessels. Mast cells stained with TB showed variably expressed gamma-metachromasia, which was best visible in those situated in the lamina propria of the mucosa. The prevailing parts of mast cells, however, were AB-positive after AB-safranin staining. This was mostly found in mast cells of the tunica muscularis and in mast cells of perivascular location in the tunica adventitia. Immunocytochemically, mast cells were found to be positive for histamine and vasoactive intestinal polypeptide in the muscle coat, and to histamine in the adventitia, as well. On the basis of obtained results it was presumed that the mast cells in porcine ureter most probably took part not only in keeping of local homeostasis, but played also an important role of mobility of smooth muscle cells in the middle layer of ureter on one hand, and, on the other, in the adventitial blood vessels.

[A case of a papilloferous cystadenoma of boundary malignancy in a 29-year-old female patient with high surgical risk].

A case of papilloferous serous cystadenoma of boundary malignancy in a 29-year old female patient, with VCS syndrome--status post phlebothrombosis ileofemoralis dextra. Thrombosis vena cava superior is presented.

[A case of a papillary cystadenoma of boundary malignancy in a 29-year old female patient with high surgical risk].

A case of papilloferous serous cystadenoma of boundary malignancy in a 29-year old female patient, with VCS syndrome--status post phlebothrombosis ileofemoralis dextra. Thrombosis vena cava superior is presented.

[Application of Nd YAG laser for the treatment of preneoplastic lesions of the uterine cervix]. / Prilozhenie na Nd YAG laser pri terapiiata na preneoplaziite na matochnata shiika.

The authors represented the data of treatment for CIN with Nd YAG laser by 266 women Two laser methods was use--direct by 93 women and indirect by 173. The effect of treatment with direct Nd YAG laser application for all cervieaf lesions ist 100%. There was no one patients with relapse by use the second method the effect ist for 92 to 96.7%. The persistence of lesion ist 3 to 7%, and relapse--3%.