Experimental autoimmune myocarditis in rats and therapeutic histamine H1 - H4 receptor inhibition.

Myocarditis, a life threatening disease, is still not adequately treated. Histamine plays an important role in physiology and pathophysiology of cardiovascular system. All four histamine receptors (H1R - H4R), are present in the heart. Experimental autoimmune myocarditis (EAM) was used to investigate which histamine receptor had a greater impact on the disease's progression. EAM was evoked in Lewis rats by porcine myosin immunization. Mepyramine, ranitidine and ciproxifan were used to inhibit H1R, H2R and H3R receptors, respectively, and 2,4-diaminopyrimidines: ST994, ST1012, ST1006 were ligands of H4R. Quinapril, an ACE inhibitor, served as a reference drug. Drugs were administered daily, either from 0 - 2 weeks or from 2 to 4 weeks post EAM induction. Cardiac dysfunction developed with significant decreases in left ventricular ejection fraction and fractional shortening due to dilatation and wall thickening. EAM rats treated with mepyramine and ST994 in weeks 0 - 2 had the lowest decreases. These treated with ST994, ST1012 or quinapril performed much better the following 2 weeks without therapy than did the other groups. On autopsy their hearts were smaller, less fibrotic, histopathological changes in them of a lower grade. When the treatment started with 2 weeks' delay, the ST994-treated EAM rats showed the highest median survival. H4 receptor antagonism inhibits heart remodelling, preserves heart contractility, improves survival and may be of potent therapeutic relevance in human clinics. The blockade of H1 receptor inhibits heart dilatation but does not prolong the life.

Parotid gland tumors in children - pre- and postoperative diagnostic difficulties.

Major salivary gland tumors are very rare in the developmental period. Confirming tumor changes in the salivary gland requires precise diagnostic imaging involving an ultrasonography scan, computed tomography and magnetic resonance. Needle aspiration biopsy (NAB) of the tumor is of high importance. Excision is the basic treatment method in cases of parotid gland tumor. The statistical data concerning tumors favor less invasive methods, which seems logical in the population of children. The surgical methods used in tumor treatment feature extracapsular excision of tumor, partial parotidectomy and total parotidectomy, sometimes followed by lymphatic node surgery. The clinical cases presented in the paper show difficulties with pre- and postoperative histopathological diagnosis in major salivary gland tumors in children. A core biopsy of the tumor may improve the accuracy of preoperative diagnosis but it does not exclude the possibility of misdiagnosis.

Prognostic importance of cyclin E1 expression in neuroblastic tumors in children.

A number of studies have indicated that cyclin E plays an important role in a variety of neoplastic processes. In our study we evaluated cyclin E1 expression and the possible prognostic value of this protein in neuroblastic tumors in children. Cyclin E1 expression was investigated by means of immunohistochemical analysis of 25 neuroblastic tumor tissue samples. We found a significant correlation between high cyclin E1 expression and deaths due to neoplastic disease. The mean values of cyclin E1 indexes in fatal cases were twice as high as in other cases. The results indicate that high cyclin E1 expression may have prognostic importance in neuroblastic tumors in children.

Systematic administration of chloroquine in discoid lupus erythematosus reduces skin lesions via inhibition of angiogenesis.

BACKGROUND: Discoid lupus erythematosus (DLE) is a chronic cutaneous form of lupus erythematosus, characterized by inflammation and scarring skin lesions, with lymphocyte infiltration and vasodilation. Antimalarial drugs have beneficial therapeutic effects in DLE, partially resulting from their immunomodulating and photoprotective properties. The possible influence of these drugs on angiogenesis has not been previously evaluated. AIMS: To investigate the impact of chloroquine (CQ) treatment on the expression of vascular endothelial growth factor (VEGF, a major regulator of angiogenesis) and CD34 (a transmembrane glycoprotein expressed on endothelial cells and involved in tethering lymphocytes) in patients with DLE. METHODS: A 3-mm skin biopsy was taken from typical skin lesions in 10 people with DLE. Another biopsy was taken from the same area after 3 months of treatment with CQ (250 mg/day). Skin sections were stained with monoclonal antibodies directed against VEGF and CD34. The intensity of epidermal VEGF expression, and the number and area of CD34-positive dermal blood vessels were assessed. RESULTS: CQ treatment induced a reduction in epidermal VEGF expression. It also resulted in a significant decrease in the median number of CD34+ dermal blood vessels (from 219 to 125 vessels per mm(2)). Furthermore the median vessel area was significantly lowered from 9.76 x 10(6) to 6.92 x 10(6) mm(2) per mm(2) of the dermis. CONCLUSIONS: These results indicate that one beneficial effect of CQ treatment in DLE may be due to its antiangiogenic properties.

Chloroquine treatment reduces the number of cutaneous HLA-DR+ and CD1a+ cells in patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that can be exacerbated by exposure to ultraviolet radiation (UVR). The number and phenotype of antigen presenting cells in the skin play a role in cutaneous immune response generation. Although antimalarials are widely used in SLE treatment, their mode of action is not completely elucidated. The aim of our study was to determine the effect of chloroquine treatment on HLA-DR+ and CD1a+ cell number in locally irradiated (three minimal erythema doses of UVB) and normal appearing skin in SLE patients and healthy subjects. A significantly higher number of HLA-DR+ and CD1a+ cells were found in both locations in SLE patients compared with controls. Following three months of daily chloroquine treatment (250 mg), the HLA-DR+ and CD1a+ cell counts were significantly reduced in both irradiated and unirradiated sites of SLE patients, although still higher than in controls. Chloroquine treatment reduces the number of antigen presenting cells in the skin of SLE patients, and this effect may explain the antimalarials beneficial immunoregulatory and anti-inflammatory properties.

Prognostic value of proliferating cell nuclear antigen in Wilms' tumour in children.

AIMS: To evaluate the prognostic value of index Proliferating Cell Nuclear Antigen (PCNA) in Wilms' tumour in children. METHODS: The study comprised 64 children aged from 2 days to 13 years treated according to the SIOP (Society International of Oncology Paediatric) and accepted by the PPGGL (Polish Paediatric Group for the Treatment of Solid Tumours). The studies were conducted on tumour tissue removed during surgery, fixed in formalin and embedded in paraffin blocks. Sections (4 microns) were evaluated by immunohistochemistry, using the peroxidase method to determine the expression of PCNA in Wilms' tumour cells by primary monoclonal antibody NCL-PCNA from Novocastra. RESULTS: The percentage of immunopositive cells in particular fragments of the tumour ranged from 0--93%, mean 30.5%, median 25.5%. Mean and median values enabled division of children into two groups: Group A, where the percentage of cells staining with anti-PCNA was <30% and Group B, where this percentage was >30%. The expression of PCNA was evaluated in various stages of advancement, various histological types and depending on the course of disease. The studies revealed the correlation between index PCNA and stage of advancement P<0.01, index PCNA and histological type of Wilms' tumour P<0.025. Moreover we observed that deaths were found more frequently in tumours with index PCNA >30%, P<0.001. CONCLUSIONS: PCNA is a useful prognostic factor in Wilms' tumour in children.

Prognostic value of p53 expression in Wilms' tumor in children.

BACKGROUND: The aim of this report was to evaluate the prognostic value and clinical correlations of p53 expression in children with Wilms' tumor. MATERIAL AND METHODS: The study comprised 61 children aged from 2 days to 13 years (median 39 months), diagnosed and treated according to SIOP and PPGGL criteria in three centers co-operating with the PPGGL. The studies were conducted on tumor tissue removed during surgery, fixed in formalin and embedded in paraffin blocks. Then 4-micron sections were evaluated by immunohistochemistry, using the peroxidase method to determine the expression of p53 in Wilms' tumor cells by means of primary monoclonal antibody NCL-p53 from Novocastra. RESULTS: The percentage of immunopositive cells in particular fragments of the tumor ranged from 0% to 70% (mean 20.4%, median 16.0%). The mean and median values enabled the children to be divided into two groups: Group A, where the percentage of cells staining with anti-p53 antibody was >20% (23 cases), and Group B, where this percentage did not exceed 20%. The expression of p53 was then evaluated in various stages of advancement and various histological types, depending on the course of the disease. In Group A, tumors at higher stages of advancement stages were more frequent (p<0.05), and showed a higher degree of malignancy (p<0.06; EFS=56.53%). In Group B, lower stages of advancement were more frequent (p<0.05), the degree of malignancy was lower, and the EFS was 81.58%. A discrimination test, however, showed that the determination of p53 expression in Wilms' tumor cells has moderate sensitivity (58.825%), positive prediction (43.47%), and relatively high specificity (70.45%) and negative prediction (81.57%), which means that low indexes of p53 expression have higher prognostic value. CONCLUSIONS: The index of p53 expression is not an independent prognostic factor in Wilms' tumor in children, but this determination may be helpful in identifying high-risk and low-risk patients.

Cherubism--clinical picture and treatment.

UNLABELLED: Cherubism is a rare, painless, disfigurating disease primarily affecting bones of the jaws. OBJECTIVE: To report on five patients with cherubism. The symptoms of the disease, methods of management and possible mode of inheritance are discussed and literature is reviewed. PATIENTS: The study involves five cherubs, members of one family. The diagnoses were based on history, physical examination, laboratory tests, X-ray parameters, and clinical follow-up. One member of the family had surgical intervention. The remaining cases were left for observation. RESULTS: Good aesthetic and long lasting effect was reached in the operated patient. CONCLUSIONS: Treatment is unnecessary unless functional or emotional disturbances develop. An autosomal recessive pattern of inheritance is suggested for these cases, although autosomal dominant transmission has been previously established.

[Significance of angiogenesis in laryngeal cancer]. / Wartosc oceny angiogenezy w raku krtani.

Angiogenesis as multistage process, which is required for forming of new vessels and is essential for the growth of solid tumors and making metastasis. Sixty patients with laryngeal cancer were analyzed. They were treated in ENT Department of Medical University in Lodz and follow-up period was not shorter than 5 years. Intensity of neovascularisation was examined by means of immunohistochemical techniques carried out on paraffin sections using selected antibody against antigens of vessel cells. The value of the prognostic factors of angiogenesis intensity were evaluated against: dissected nodal metastasis, tumor recurrences and metastasis observed in prospective follow-up and after cancer treatment live time. Morphological properities of tumor or hospital treatment of the patients with laryngeal cancer were analyzed as well. Taking into consideration the results, we can make the conclusion that investigation of angiogenesis can be forthcoming prognostic factor for the laryngeal cancer.

[Assessment of cell proliferation antigen Ki-67, protein p53 related to apoptosis and angiogenesis in laryngeal cancer]. / Ocena ekspresji antygenu proliferacji komórkowej Ki-67, bialka zwiazanego z apoptoza p-53 i intensywnosci angiogenezy w raku krtani.

Kinetics (growth fraction of tumour cell populations), death process of cancer cells (apoptosis and necrosis) and neovascularisation in tumour (angiogenesis) have influence on the growth of cancer. Sixty patients with laryngeal cancer treated in ENT Department of Medical Academy of Lodz were analysed. Proliferation activity of cancer cells was examined by means of selection appropriate antigen (Ki-67) characteristic for cell cycle utilising immunohistochemical techniques carried out on laryngeal cancer paraffin samples. Expression of selected protein connected with apoptosis (p-53) and intensity of angiogenesis were examine using selected antibody (anti-CD34) aimed against epithelial antigens. Above-mentioned markers were correlated with: stage of cancer progression, recurrences and metastasis of laryngeal cancer and follow-up of the patients. The morphological properties were examined as well. The researches on apoptosis, angiogenesis and proliferation of cancer cells can be used as prognostic factors for the patients with laryngeal cancer.

[Programmed cell death research in laryngeal cancer]. / Badania nad programowana smiercia komórki w raku krtani.

Apoptosis--the programmed sell death is the process of characteristic events on morphological, biochemical and molecular level which lead consequently to cell death. This process require activation of some genes i.e. p-53, mdm2 and inhibiting others i.e. bcl-2. Sixty patients with laryngeal cancer treated in ENT Department of Medical Academy of Lodz were analysed. Expression of the p-53 and bcl-2 genes' products was examined by means immunohistochemical techniques carried out on laryngeal cancer paraffin samples. Above-mentioned markers were correlated with: stage of cancer progression, recurrences and metastasis of laryngeal cancer and follow-up of the patients. Initial results indicate the possible utilisation of apoptosis as prognostic factors for the patients with laryngeal cancer.

[Evaluation of CD44 adhesion molecule, nm23 gene product expression and intensity of angiogenesis in patients with laryngeal cancer]. / Ocena ekspresji czasteczki adhezyjnej CD44, produktu genu nm23 i intensywnosci angiogenezy u chorych z rakiem krtani.

One of the most important factor in prognosis of the patients with laryngeal cancer is presence of the metastases in lymph nodes of the neck. The main purpose of the paper was the evaluation of CD34 and FVIII antigens as angiogenesis markers, and nm23 protein and CD44 antigen expression as metastasis potential markers and description of their role in the tumour progression and making metastasis in the patients with laryngeal cancer. Paraffin-embedded tissue sections from 89 patients with laryngeal cancer were stained with a monoclonal antibody raised against CD34 and FVIII antigens, against nm 23 protein and against CD44 antigen. Measuring the density of the microvasculature in tumour was investigated. We found significant dependence between intensity of angiogenesis (IA) and pT, nodal metastasis, histological grading and survival. There were also significant correlation between nm23 protein expression and nodal metastasis, and between CD44 antigen expression and pT, nm23 protein expression and FVIII antigen expression. Evaluation of mentioned markers allowed to asses the aggressiveness of tumour cells and anticipate neck metastasis in the patients with laryngeal cancer.

[Expression of selected markers for apoptosis, proliferation and metastasis in evaluation of laryngeal cancer invasiveness dynamics]. / Ekspresja wybranych wskazników apoptozy, proliferacji i przerzutowosci w ocenie dynamiki rozrostu raka krtani.

Evaluation of the biology of laryngeal cancer cell is connected either with many process inside the cell or reactions between cancer cell itself and extracellular matrix. The main purpose in this paper was the evaluation of p53 protein, bcl-2 protein, Ki-67 antigen and CD44 adhesive molecule expressions in comparison to clinical and histopathological features in patients with laryngeal cancer. Paraffin-embedded tissue sections from 89 patients with laryngeal cancer were stained with a monoclonal antibody raised against p53 and bcl-2 proteins, Ki-67 and CD44 antigens using a peroxidase-labelled streptavidin-biotin kit. There were statistically significant relationships between p-53 protein over-expression and pT, histological grading, survival and Ki-67 and CD44 antigens expressions. There were no correlation between bcl-2 protein expression and clinical and histopathological features. We observed statistically significant correlation between Ki-67 expression and pT, histological grading, recurrences and survival. Expression of CD44 statistically significant correlated only with tumour size. We conclude that comparison of data covering mentioned tumour markers expression gives valuable evaluation of biological activity of cancer cells and may allow to create the immunological panel of tumour markers which simplify the prognosis about nodal metastases, recurrences and survival in patients with laryngeal cancer.

Gastrointestinal sutureless anastomosis in pigs using absorbable intraluminal stents, stent placement devices, and fibrin glue - a summary.

INTRODUCTION AND METHODOLOGY: To develop a practical gastrointestinal sutureless anastomosis technique, 164 end-to-end and end-to-side anastomoses were performed on the small intestine (SI), large intestine (LI), rectum, esophagus and gallbladder in 109 female Landrace pigs weighing 25 kg and 35 kg. There were 116 fibrin glue (FG) and 48 sutured anastomoses. The end-to-end SI and LI anastomoses were divided into five groups: sliding absorbable intraluminal nontoxic stent (SAINT); SAINT placement device (SAINT-PD); nonsliding SAINT (nST); sutureless stapler (SS); and sutured controls. The SAINT had a sucrose base, with some having reinforcing fibers. RESULTS: No deaths from dehiscence occurred in any group except one FG-cylinder attempt in the colon (technique abandoned). Statistical analysis of gross pathology indices showed no significant group differences. However, trends favored the SAINT in many indices, including grade-0 intraluminal tissue ridge formation (70.8% SI, 84.4% LI) and grade-0 adhesion rates (45.8% SI, 73.1% LI). Histologic examination showed fewer giant cells, less inflammation, less scar tissue formation and faster healing in the SAINT and nST anastomoses than controls. Follow-up of 300-540 days demonstrated no signs of necrosis or stenosis in the SAINT anastomoses. The nST had excellent results; however, it seems impractical in SI anastomoses and unsuitable for LI. CONCLUSIONS: The SAINT-PD has potential for all gastrointestinal sites, but needs larger experimental trials. The SS technique is impractical and had high tissue ridge formation and adhesion rates. These preliminary trials suggest the simplicity, versatility and safety of the SAINT technique; however, the small groups limit result interpretation. The results present a starting point for sutureless FG gastrointestinal anastomosis, and future experimental evaluation with more extensive statistical analyses in larger studies are needed.

Relationship between passive smoking, recurrent respiratory tract infections and otitis media in children.

UNLABELLED: The cause of upper respiratory tract infections (URTI) are multifactorial (enlarged adenoid, environmental conditions, staying at the care centers, smoking parents, allergy). Directly, viral infection causes damage to the ciliary cells and mucociliary clearance in the nasopharynx and Eustachian tube, promotes tubal occlusion and provokes otitis media. Enlarged adenoids reduce ventilation to the nasopharynx, increase accumulation of the secretion and provide a good condition for bacteria. AIMS OF THE STUDY: Evaluation of the factors playing a role in recurrent URTI and otitis media in children. Clinical and histopathological examination of adenoid tissue of children who were passive smokers and children who were not exposed to cigarette smoke. Evaluation of the difference between ciliary-mucous transport among passive smokers and children not exposed to cigarette smoke. METHODS: The analysis of interview questionnaires in 1000 children aged 3-14 years. Histopathological examinations of adenoid tissue excised in the group of children of recurrent upper respiratory tract infections and serous otitis media exposed and not exposed to cigarette smoke. CONCLUSIONS: Among the risk factors for URTI, the most important are: (1) socio-economic conditions; (2) staying at day care centers; and (3) passive smoking. Allergy was confirmed in 35-38% of URTI children. Surgical treatment was undertaken in 11.4-32.5% of URTI children (tonsilloadenoidectomy). Histopathological and ultrastructural evaluation of adenoid tissue in passive smoking children indicates significant differences to children not exposed to cigarette smoke.

Immunohistochemical evaluation of tumour angiogenesis in adenocarcinoma and squamous cell carcinoma of lung.

Experimental studies revealed that growth and expansion of solid tumours depend on angiogenesis. Angiogenesis is very important factor for neoplastic metastasis. The presence of the metastasis is an ominous prognostic factor for many tumours, also for lung cancer. Studies of tumour microvessel density in resected non-small lung cancers have not given convincing data about value of angiogenesis. Only few reports regarded the association with angiogenesis in different histological types in lung carcinoma. Samples of 35 adenocarcinomas and 41 squamous cell resected, primary lung carcinomas were studied. Paraffin sections of tumours were stained immunohistochemically by antibody against endothelial marker CD34. Angiogenesis intensity was measured in the areas of the most active fields of tumour neovascularization. Microvessel density (MD) was higher in adenocarcinoma comparing to squamous cell cancer, but the difference was not statistically significant (p = 0,095). The groups of various stage of extension of disease in each histological type were compared-MD correlated with lymph node metastasis (p = 0,003) in the adenocarcinoma, whilst in squamous cell can cer differences between various groups of nodal involvement were not statistically significant (p = 0,53 and p = 0,22 respectively). Our results suggest that more intensive angiogenesis in adenocarcinoma could be more important factor for metastasis of adenocarcinoma than for squamous tumours. In the latter group angiogenesis may be more important for growth of squamous cell cancers, while the spread of squamous tumours may depend on other mechanisms.

Sliding, absorbable, reinforced ring and an axially driven stent placement device for sutureless fibrin glue gastrointestinal anastomosis.

Reduced blood flow of from 43 to 71% has been reported in sutured and stapled anastomoses. The sutureless sliding, absorbable, intraluminal, nontoxic stent (SAINT)-fibrin glue anastomotic method, which clamps the stump margins between 2 dissolving surfaces, includes only two stages of temporary compression (about 6 min total using 4 IU/mL thrombin) during the glue application in order to promote vascularization. A SAINT placement device (SAINT-PD) was introduced to facilitate low rectal anastomoses. Morphohistologic results from limited trials using fibrin glue with an untied sutureless stapler technique and a prototype non-gear-driven SAINT-PD, neither having the two dissolvable clamping surfaces of the SAINT, showed a 29 and 25% incidence of intraluminal tissue ridges, respectively. Since these tissue ridges could result in subclinical dilatation or frank stenosis, and the more extensive SAINT trials had an 8% incidence of tissue ridges, redesign of the SAINT-PD was undertaken. Consequently, to improve the anastomotic quality of the SAINT-PD, the sliding absorbable reinforced ring (sucrose base) acting as the second dissolvable surface for the SAINT-PD and a new axially controlled geared SAINT-PD design are described.

Gastrointestinal sutureless anastomosis using fibrin glue: reinforcement of the sliding absorbable intraluminal nontoxic stent and development of a stent placement device.

Sutureless anastomosis of the gastrointestinal tract using fibrin glue and sliding absorbable intraluminal nontoxic stents (SAINTs) has two shortcomings, stent shaft breakage and the lack of a transanal insertion device (TID) for low anterior resection. Reinforcement of the sucrose base SAINT (R-SAINT) is described. Sutureless anastomosis is attempted using a stapleless mechanical stapler (SS) and used as preprototype to screen histologically and mechanically for TID anastomoses in the small intestine. Finally, a prototype absorbable head SAINT placement device (SAINT-PD) intended for TID, similar to the SS, is utilized on the small intestine. Fifty-seven Landrace pigs weighing 25-35 kg were used to perform 58 anastomoses, including the small intestine (15 manual, 19 SAINT, 11 SS, 5 R-SAINT, 6 SAINT-PD) and large intestine (2 R-SAINT). All anastomoses performed with the R-SAINT succeeded on the first attempt even if the shaft cracked. The SS technique proved impractical, but the histological screen results from 7 to 60 days did approximate those of corresponding SAINT anastomoses. The SAINT-PD demonstrated operational improvement over the SS, but the histological results were similar to both the SS and SAINT. The advantages of the R-SAINT and SAINT-PD are that they leave no foreign bodies or pressure clamping devices at the anastomostic site. Larger studies may show the R-SAINT and the SAINT-PD to be practical, new surgical tools in sutureless fibrin glue anastomosis.