Changes in spinal curve during dentistry studies measured with a Spinal Mouse device: A five-year prospective study.

INTRODUCTION: Musculoskeletal disorders (MSDs) often arise and develop during dentistry studies. The most affected regions are related to the spine. Possible associations between spinal curve parameters and MSDs have not yet been investigated amongst dentistry students. This longitudinal observational study aimed to determine whether spinal curve changes during dentistry studies, analyse the relationship between objective findings and subjectively declared MSDs and compare spinal curve parameters with those published in the literature. MATERIALS AND METHODS: Seventy-three dentistry students answered a questionnaire on MSDs, and were examined using the Spinal Mouse® device at the beginning, in the middle, and at the end of their 5-year study. RESULTS: The spinal curve exhibited a gender diversity in the lumbar lordosis angle, sacrum inclination, and thoracolumbar ratio. From the first to fifth study year, we observed an increase in the range of motions in the sagittal and frontal planes, an increase in the maximal extent of right lateral inclination, and a decrease in maximal left lateral inclination. Whole-spine backward inclination increased only in women, and forward sacral inclination decreased. No statistically significant relationships were found between the objective findings and subjectively declared MSDs. CONCLUSIONS: The spinal curve shape differed between men and women and changed during dentistry studies. No objective markers or predictors of MSDs were found amongst the dentistry students. These findings can serve as a benchmark for further studies on the association between MSDs and objective findings.

DNA hypermethylation of tumor suppressor genes TWIST1, GATA4, MUS81 and NTRK1 in endometrial hyperplasia.

OBJECTIVE: To investigate DNA methylation of specific tumor suppressor genes in endometrial hyperplasia compared to normal endometrial tissue. File and methodology: To search for epigenetic events, methylation-specific multiplex ligation-dependent probe amplification was employed to compare the methylation status of 40 tissue samples with atypical endometrial hyperplasia, 40 tissue samples with endometrial hyperplasia without atypia, and 40 control tissue samples with a normal endometrium. RESULTS AND CONCLUSION: Differences in DNA methylation among the groups were found in TWIST1, GATA4, MUS81, and NTRK1 genes (TWIST1: atypical hyperplasia 67.5%, benign hyperplasia 2.5%, normal endometrium 22.5%; P < 0.00001; GATA4: atypical hyperplasia 95%, benign hyperplasia 65%, normal endometrium 22.5%; P < 0.00001; MUS81: atypical hyperplasia 57.5%, benign hyperplasia 22.5%, normal endometrium 5%; P < 0.00001; NTRK1: atypical hyperplasia 65%, benign hyperplasia 27.5%, normal endometrium 10%; P < 0.00001). Higher methylation rates were observed for the tumor suppressor genes of TWIST1, GATA4, MUS81, and NTRK1 in samples with atypical endometrial hyperplasia compared to samples with normal endometrial tissue, and higher methylation rates were found in samples with atypical endometrial hyperplasia compared to samples of benign endometrial hyperplasia. DNA methylation of TWIST1, GATA4, MUS81, and NTRK1 is involved in the pathogenesis of atypical endometrial hyperplasia.

Sex identification in Australian skinks (Egernia stokesii, E. hosmeri, E. striolata) using cloacoscopy.

Thirteen adult healthy captive skinks, six Gidgee spiny-tailed skinks (Egernia stokesii), three Hosmer's spiny-tailed skinks (Egernia hosmeri) and four tree crevice-skinks (Egernia striolata), were submitted to the study. The weight of the animals ranged from 28 g to 146 g. All skinks were explored during their putative mating season, which is December. Lizards were restrained manually and positioned in ventral recumbence. Cloacoscopy was performed with the rigid endoscope, protecting channel, endocamera and recording system Telepack-Pal (Karl Storz Endoskope, Tuttlingen, Germany). While the endoscope was gently introduced into the cloaca and advanced into the coprodeum sterile saline was flushed into the cloaca through the protecting channel. The endoscope was then slowly withdrawn to allow visualization of the main structures of the urodeum and proctodeum. Male skinks are characterized by the presence of urethral papillae and only one horizontal septum which divides the chamber of the urodeum into two subchambers. Female skinks are characterised by the presence of two septa. The central-dorsal fold that divides the urodeum into two pouches in female skinks is absent in males. Cloacoscopy proved an effective method of sex identification and can be considered a valuable method for breeding and conservation in monomorphic skink lizards.

Malignant ovarian dysgerminoma in a 16-year-old leopard gecko (Eublepharis macularius).

The 16-year-old female leopard gecko (Eublepharis macularius) was presented with distended coelom and cachexia. Examination of the faecal sample ruled out the presence of protozoan parasites. A radiographic examination confirmed the presence of radiopaque foreign material in the intestine. The conservative treatment with tramadol, butylscopolamine, famotidine, vitamin B complex, and supportive fluid therapy with Hartmann solution and Duphalyte, was performed for 14 days. Ultrasonographic examination revealed the presence of a large mass adherent to the liver (with hypoechoic regions), a thin-walled cystic structure close to the liver, and coelomic effusion. Surgical exploration revealed a large mass on the right ovary. The unilateral (right) ovariectomy was performed. Histologic examination of the mass revealed dysgerminoma with an invasion of the ovarian bursa and blood vessels. Nine months after the surgery the patient was active and doing well. In reptiles, dysgerminoma is an uncommon type of neoplasia. To the best of our knowledge, this is the first case of dysgerminoma tumour diagnosed intravitally and treated successfully in lizards.

Effect of GnRH agonist (deslorelin) on reproductive activity in captive female veiled chameleons (Chamaeleo calyptratus).

Eighteen 5 months old veiled chameleon females (Chamaeleo calyptratus) were used in the study. Seven females received subcutaneous implants with gonadotropin-releasing hormone agonist (GnRH) deslorelin acetate whereas eleven females were used as control animals without any implants. Females were kept in five terraria, in groups of four females (in 3 terraria) and groups of three females (in 2 terraria), respectively. A minimum of one female with GnRH implants was present in each terrarium. They were kept under standard husbandry conditions. Females of both groups (females with GnRH and controls, respectively) were monitored for three years. No differences between females with GnRH implants and females without GnRH implants were observed with respect to the presence of large ovarian follicles, number of eggs/female, or number of clutches/female. GnRH implants did not prevent spontaneous reproductive activity in any of the 7 females with implants. Ovariectomy was performed in 8 females (4 females with GnRH implants and 4 control females). Ten females (3 females with GnRH implants and 7 control females) had to be euthanised. In 17 of 18 female veiled chameleons of this study histologic examination revealed heterophilic granulomatous oophoritis. The use of GnRH agonist implants did not prove to be an appropriate method for the control of reproductive function in captive female veiled chameleons.

Plasma bile acids in healthy green iguanas and iguanas with chronic liver diseases.

The aim of the study was to establish reference values for plasma bile acid (BA) concentrations in a collection of healthy green iguanas and to compare the results with BA concentrations in iguana patients presented to the clinic with various types of chronic liver diseases, patients with other chronic diseases and healthy iguanas that were presented for routine or pre-surgical health check-up. The concentration of BA was determined using the enzymatic colorimetric method. Mean plasma bile acid concentration in 110 samples from healthy green iguanas fasted for 24 h was higher (15.89 ± 15.61 µmol/l) than plasma bile acid concentration in the same iguanas fasted for 48 h (9.56 ± 8.52 µmol/l) (P < 0.01). The 3α-hydroxy bile acid concentration was significantly altered in 9 patients suffering from chronic liver diseases (diagnosed by histology) (84.85 ± 22.29 µmol/l). BA concentration in one iguana with hepatocellular adenoma (13.0 µmol/l) was within the interval of BA in healthy iguanas. Mean plasma BA concentration in 10 green iguanas that were suffering from various types of chronic diseases, but without any hepatopathy was 7.85 ± 4.86 µmol/l. The mean plasma BA concentration in 18 samples from green iguanas presented to the clinic for routine health check-ups and 17 green iguana females with preovulatory follicle stasis (POFS) syndrome presented for ovariectomy was 11.95 ± 9.43 µmol/l and 12.97 ± 9.06 µmol/l, respectively. The data collected from this study suggest that plasma bile acids are significantly increased in green iguanas suffering from chronic liver diseases.

Differences in the anatomy of the lower respiratory tract in selected species of the order Testudines.

The lower airways of turtles consist of the larynx, trachea and bronchi. Due to incomplete information about the anatomical structure of the lower respiratory tract in turtles, we decided to explore this topic more deeply. The material for this study included 95 turtle cadavers. Terrestrial tortoises were represented by 63 individuals from 8 species, aquatic and semi-aquatic turtles by 32 individuals from 11 species. The sex ratio was 54 females to 41 males. In addition to the anatomical autopsies and assessments of the macroscopic structures, numerous measurements were performed. The length, width and height of the larynx and trachea, and the length of the bronchi were measured. We found clinical significance in two parameters. The first one is the formation of the rima glottidis, which is significantly shorter and narrower in the aquatic turtles than in the terrestrial tortoises. It follows that for these species, we should use a smaller diameter of endotracheal cannula for intubation. The second parameter is a very short trachea in tortoises of the genus Testudo. The length of the trachea is only a few centimetres, which significantly increases the risk of intubation into one bronchus only.

Congestive heart failure in a veiled chameleon (Chamaeleo calyptratus): A case report.

A two-year-old male veiled chameleon (Chamaeleo calyptratus) was referred for a gular oedema and bilateral blepharoedema. The echocardiography revealed a ventricular hypertrophy, pericardial effusion, and valvular regurgitation of the right atrioventricular valve. Treatment with hydrochlorothiazide, enalapril, and carvedilol was commenced. Within 3 weeks of treatment, the valvular regurgitation was noticeably decreased. In the 4th week of treatment, the echocardiography revealed a reduction in the myocardium hypertrophy. After an additional month of home treatment, the patient was presented with anorexia and decreased activity. Despite the supportive care, the patient died. The histopathology revealed mild to moderate fibrosis of the epicardium. Moderate to severe fibrosis, degeneration of the myofibrils, fatty atrophy, interstitial oedema and mild calcification was seen in the atria. The tunica intima, media and adventitia of the major cardiac vessels were moderately fibrotic, swollen and interfused by myxoedema. The kidney histopathology revealed moderate sclerosis and atrophy of the glomeruli, vacuolation of the tubular epithelium, fibrosis, and infiltration of the leucocytes in the interstitium. The therapeutic protocol with hydrochlorothiazide, ACE inhibitor enalapril and ß-blocker carvedilol reduced the myocardium hypertrophy and the valvular regurgitation; however, the prolonged use of diuretics jeopardized the renal function in our patient. Frequent blood analyses are necessary using diuretics in reptile patients.

Analysis of Results of Specific IgE in 100 Atopic Dermatitis Patients with the Use of Multiplex Examination ALEX2-Allergy Explorer.

BACKGROUND AND AIM: Progress in laboratory diagnostics of IgE-mediated allergy is the use of component-resolved diagnosis. Our study analyses the results of specific IgE to 295 allergen reagents (117 allergenic extracts and 178 molecular components) in patients suffering from atopic dermatitis (AD) with the use of ALEX2 Allergy Explorer. METHOD: The complete dermatological and allergological examination, including the examination of the sensitization to molecular components with ALEX2 Allergy Explorer testing, was performed. The statistical analysis of results was performed with these methods: TURF (total unduplicated reach and frequency), best reach and frequency by group size, two-sided tests, Fisher's exact test, and chi-square test (at an expected minimum frequency of at least 5). RESULTS: Altogether, 100 atopic dermatitis patients were examined: 48 men, 52 women, the average age 40.9 years, min. age 14 years, max. age 67 years. The high and very high level of specific IgE was reached in 75.0% of patients to 18 molecular components: from PR-10 proteins (Aln g 1, Bet v 1, Cor a1.0103, Cor a1.0401, Fag s 1), lipocalin (Can f 1), NPC2 family (Der f 2, Der p 2), uteroglobin (Fel d 1), from Alternaria alternata (Alt a 1), Beta expansin (Lol p 1, Phl p 1), molecular components from Timothy, cultivated rye (Secc pollen) and peritrophin-like protein domain Der p 23. The high and very high level of specific IgE to other lipocalins (Fel d 7, Can f 4), to arginine kinase (Bla g 9, German cockroach), and to allergen extracts Art v (mugwort), and Cyn d (Bermuda grass) reached 52.0% of patients. The severity of AD is in significant relation to the sensitization to molecular components of storage mites (Gly d 2, Lep d 2-NPC2 family), lipocalins (Can f 1, Can f 2, Can f 4, and Can f 6), arginine kinase (Asp f 6, Bla g 9, Der p 20, Pen m 2), uteroglobin (Fel d 1, Ory c 3), Mn superoxide dismutase (Mala s 11), PR-10 proteins (Fag s 1, Mal d 1, Cor a 1.0401, Cor a 1.0103), molecular components of the peritrophin-like domain (Der p 21, Der p 23), and to Secc pollen. In the subgroup of patients suffering from bronchial asthma, the significant role play molecular components from house dust mites and storage mites (Lep d 2, Der p 2, Der f 2-NPC2 family), cysteine protease (Der p 1), peritrophin-like protein domain (Der p 21, Der p 23), enolase from Alternaria alternata (Alt a 6), and Beta expansin Phl p 1. CONCLUSION: The results of our study demonstrate the detailed profile of sensitization to allergens reagents (allergen extract and molecular components) in patients with atopic dermatitis. We show the significance of disturbed epidermal barrier, resulting in increased penetration of allergens. We confirmed the significant relationship between the severity of AD, the occurrence of bronchial asthma and allergic rhinitis, and high levels of specific IgE to allergen reagents. Our results may be important for regime measures and immunotherapy; Der p 23 shall be considered as an essential component for the diagnosis and specific immunotherapy of house dust mite allergy.

Quality of life of patients with psoriasis.

OBJECTIVE: The two main aims of our study were to assess the quality of life (QOL) of patients with moderate or severe psoriasis treated by Goeckerman therapy and to compare QOL of our patients on the date of admission and 1 month after their discharge. METHODS: We performed a prospective study on 51 patients treated for chronic plaque psoriasis by Goeckerman regimen (GR). The psoriasis area involvement and severity was measured using the Psoriasis Area and Severity Index (PASI). Patients' health-related QOL was evaluated using Psoriasis Disability Index (PDI). RESULTS: In our study we demonstrated that the QOL of this sample of 51 patients with severe forms of psoriasis was relatively good; an average PDI value on the day of admission was 9.02, which represents a small effect on patient's QOL. We did not find any significant differences in QOL between younger and older respondents or between men and women. Despite good treatment efficiency, we did not find improvement in QOL 1 month after discharge. CONCLUSIONS: The QOL of our patients was relatively good. This could be explained by the fact that our patients have been suffering from psoriasis for many years and have probably adapted to their disease. QOL of our patients was not directly dependent on the extent and intensity of skin changes. Further studies on the connection between psoriasis therapy and patients' QOL are still highly needed.

Effects of Amlodipine on Bone Metabolism in Orchidectomised Spontaneously Hypertensive Rats.

Spontaneously hypertensive rats (SHR) represent a model of essential hypertension. We studied the effect of amlodipine (AML) on bone markers, bone mineral density (BMD), and biomechanical properties of osteopenic bone induced by orchidectomy in male SHR. Rats were allocated to 3 groups and were sacrificed after 12 weeks: sham-operated control; orchidectomised control; and orchidectomised receiving a diet supplemented with AML. Indicators of bone turnover were assessed in bone homogenate, BMD was measured by dual energy X-ray absorptiometry, and the femurs were subjected to biomechanical testing. Long-term AML administration does not have a negative impact on bone metabolism and density in male SHR.

Observational cohort study of hepatic encephalopathy after transjugular intrahepatic portosystemic shunt (TIPS).

Introduction and Aim: Hepatic encephalopathy (HE) is a common complication of transjugular intrahepatic portosystemic shunting (TIPS). It is associated with a reduced quality of life and poor prognosis. The aim of this study was to compare two groups of patients who did and did not develop overt HE after TIPS. We looked for differences between these groups before TIPS. MATERIALS AND METHODS: A study of 895 patients was conducted based on a retrospective analysis of clinical data. Data was analyzed using Fisher's exact test, Chi-square, Mann Whitney test, unpaired t-test and logistic regression. After the initial analyses, we have looked at a regression models for the factors associated with development of HE after TIPS. RESULTS: 257 (37.9%) patients developed HE after TIPS. Patients' age, pre-TIPS portal venous pressure, serum creatinine, aspartate transaminase, albumin, presence of diabetes mellitus and etiology of portal hypertension were statistically significantly associated with the occurrence of HE after TIPS (p < 0.01). However, only the age, pre-TIPS portal venous pressure, serum creatinine, presence of diabetes mellitus and etiology of portal hypertension contributed to the regression model. Patients age, serum creatinine, presence of diabetes mellitus and portal vein pressure formed the model describing development of HE after TIPS for a subgroup of patients with refractory ascites. CONCLUSION: we have identified, using a substantial sample, several factors associated with the development of HE after TIPS. This could be helpful in further research.

Elution kinetics of vancomycin and gentamicin from carriers and their effects on mesenchymal stem cell proliferation: an in vitro study.

BACKGROUND: Musculoskeletal infections remain a major complication in orthopedic surgery. The local delivery of antibiotics provides the high levels required to treat an infection without systemic toxicity. However, the local toxicity of antibiotic carriers to the mesenchymal stem cells, as a result of both the peak concentrations and the type of carrier, may be significant. METHODS: To address this concern, the elution kinetics of vancomycin and gentamicin from several commercially available antibiotic carriers and several carriers impregnated by a surgeon (10 ml of each sterile carrier were manually mixed with a 500 mg vancomycin and an 80 mg gentamicin solution, and the duration of impregnation was 30 min) were assessed. Moreover, the effects of these antibiotic carriers on stem cell proliferation were investigated. The following two types of stem cells were used: bone marrow and dental pulp stem cells. RESULTS: The high eluted initial concentrations from antibiotic impregnated cancellous allogeneic bone grafts (which may be increased with the addition of fibrin glue) did not adversely affect stem cell proliferation. Moreover, an increased dental pulp stem cell proliferation rate in the presence of antibiotics was identified. In contrast to allogeneic bone grafts, a significant amount of antibiotics remained in the cement. Despite the favorable elution kinetics, the calcium carriers, bovine collagen carrier and freeze-dried bone exhibited decreased stem cell proliferation activity even in lower antibiotic concentrations compared with an allogeneic graft. CONCLUSIONS: This study demonstrated the benefits of antibiotic impregnated cancellous allogeneic bone grafts versus other carriers.

Experience of Czech Children with Alcohol Consumption in Relation to Selected Family Indicators.

OBJECTIVE: The aim of this study is to monitor and analyze the influence of chosen family indicators related to alcohol consumption and experience of drunkenness in a representative sample of Czech children. METHODS: Data for our work were obtained from the Health Behaviour in School Aged Children (HBSC) study - a World Health Organization (WHO) cross national study. The sample consisted of 4,293 children aged 11, 13 and 15 years. Data collection was conducted in June 2010 in 88 randomly selected schools in the Czech Republic using standardized questionnaires. Statistical analysis was performed in the program NCSS 9, methods of descriptive statistics, χ2 test of independence in the contingency tables and logistic regression analysis were used. RESULTS: Regular alcohol consumption (at least weekly) was reported by 7.4% of children aged eleven, 19.3% of children aged thirteen and 38.4% of children aged fifteen years. Drunkenness at least twice in their life was admitted by 3% of children aged eleven, 15% of children aged thirteen and 43% of children aged fifteen years. Both alcohol consumption and drunkenness were significantly associated (p<0.001) with age and gender. Formal structure of family was found to have significant effect on the experience of drunkenness, but no effect on alcohol consumption was recorded. Based on our results, neither the amount of time spent together, nor the children communication with parents had a statistically significant influence on alcohol consumption or drunkenness experience within each family type. However, statistically significant differences were observed between different family types (p<0.001). CONCLUSIONS: These findings indicate a high degree of liberalism of Czech society towards alcohol. It appears that alcohol consumption will remain a serious problem in Czech society, therefore, more attention should be paid to this phenomenon in the future.

The prognostic effect of neoadjuvant chemoradiotherapy on the change of PD-L1 expression in patients with locally advanced rectal adenocarcinoma.

PURPOSE: To evaluate the prognostic effect of neoadjuvant chemoradiotherapy on the change of programmed death ligand 1 (PD-L1) expression in patients with locally advanced rectal adenocarcinoma, by comparing PD-L1 expression in pretreatment biopsies and PD-L1 expression in pathological specimens after neoadjuvant chemoradiotherapy. METHODS: A total of 25 patients with rectal adenocarcinoma were evaluated. Patients were treated by neoadjuvant chemoradiotherapy (radiotherapy:44Gy normofraxionation; chemotherapy: capecitabine 825 mg/m2 in two daily doses). Surgery was performed 6-8 weeks after the chemoradiotherapy completion. PD-L1 expression was determined in endoscopic biopsies and in resected specimens with immunohistochemistry. RESULTS: All 25 patients received radiotherapy without interruption, while concomitant chemotherapy was discontinued prematurely in one patient because of hematological toxicity. In 13 patients sphincter-saving surgery were performed, and 12 patients underwent rectum resection. Downstaging was noticed in 17 patients. Stable disease was found in 5 patients, and progression in 3. The median disease free survival (DFS) was not reached. Three-year DFS was 54.3% (95% CI 34.3-74.2). The median overall survival (OS) was 60 months (95% CI 48-60). Three-year OS was 75 % (95% CI 57.7-92.3). No PD-L1 expression was noticed in pretreatment biopsy and in resected tissue after chemoradiotherapy. CONCLUSION: No prognostic effect of neoadjuvant chemoradiotherapy on the change of PD-L1 expression was demonstrated in patients with locally advanced rectal adenocarcinoma.

Chlorambucil conjugates of dinuclear p-cymene ruthenium trithiolato complexes: synthesis, characterization and cytotoxicity study in vitro and in vivo.

Four diruthenium trithiolato chlorambucil conjugates have been prepared via Steglich esterification from chlorambucil and the corresponding trithiolato precursors. All conjugates are highly cytotoxic towards human ovarian A2780 and A2780cisR cancer cell lines with IC50 values in the nanomolar range. The conjugates exhibit selectivity towards A2780 cells as compared to non-cancerous HEK293 cells, while being only slightly selective for RF24 and A2780cisR cells. In vivo, the conjugate [10]BF4 suppressed the growth of a solid Ehrlich tumor in immunocompetent NMRI mice but did not prolong their overall survival. The reactivity of the chlorambucil conjugates with glutathione, a potential target of the dinuclear ruthenium motive, and with the 2-deoxyguanosine 5'-monophosphate (dGMP-a model target of chlorambucil) was studied by mass spectrometry and NMR spectroscopy. The conjugates did not show catalytic activity for the oxidation of glutathione nor binding to nucleotides, indicating that glutathione oxidation and DNA alkylation are not key mechanisms of action. Four highly cytotoxic diruthenium trithiolato chlorambucil conjugates have been prepared. All conjugates exhibit selectivity towards A2780 cells as compared to HEK293 cells, while being only slightly active in RF24 and A2780cisR cells. In vivo, the best candidate suppressed the growth of a solid Ehrlich tumor in immunocompetent NMRI mice but did not prolong their overall survival.

In-vitro and in-vivo evaluation of the anticancer activity of diruthenium-2, a new trithiolato arene ruthenium complex [(η6-p-MeC6H4Pri)2Ru2(µ-S-p-C6H4OH)3]Cl.

In the present study, we investigated the anticancer action of the trithiolato arene ruthenium complex, [(η-p-MeC6H4Pr)2Ru2(µ-S-p-C6H4OH)3]Cl, named diruthenium-2, both in vitro and in vivo. The mechanism of antiproliferative, cytotoxic, and DNA-damaging activity, and the effect on expressions of cell cycle regulatory proteins were investigated using a WST-1-based proliferation assay, lactate dehydrogenase leakage assay, comet assay, flow cytometry, and western blot analysis. In-vivo anticancer activity was evaluated using Ehrlich tumor-bearing NMRI mice. Diruthenium-2 inhibited the growth of all cancer cell lines used, the most sensitive being gastric (AGS), breast cancer (BT-549, MCF-7, MDA-MB-231), and leukemic (HL-60, MOLT-4) cells. In MCF-7 cells, it caused a G1/S cell cycle arrest, along with an increase in the expression of protein p21 and cyclin B1. We also observed increased levels of MRN complex proteins, which, together with the results from the comet assay, indicate the formation of DNA double-strand breaks. In tumor-bearing mice, diruthenium-2 at doses of 3 and 5 mg/kg inhibits the growth of solid Ehrlich tumor, although weaker than cisplatin. However, it did not prolong the post-therapeutic survival. Our results suggest the in-vitro potential of diruthenium-2 should be further evaluated in studies using other in-vivo models.

Boldine Inhibits Mouse Mammary Carcinoma In Vivo and Human MCF-7 Breast Cancer Cells In Vitro.

Boldine is an aporphine alkaloid widely consumed in the folk medicine of some regions. Its anticancer potential has been shown but not yet elucidated. We compared the antitumor effect of orally and parenterally applied boldine in mice bearing solid Ehrlich tumor. We also explored the effects of boldine on breast adenocarcinoma MCF-7 cells in vitro. Repeated i. p. injections of 30, 60, or 90 mg boldine/kg, either alone or combined with doxorubicin, slowed tumor growth in vivo. The latter two doses also prolonged the post-therapeutic survival of the mice. When fed food supplemented with boldine at a dose of 90 mg/kg, the tumor-bearing mice survived significantly longer, but there was no effect on tumor size. Interestingly, continuous p. o. administration did not produce detectable levels of boldine in plasma or tissue samples, in contrast to high but short-lived concentrations after i. p. injections. There was neither antagonism nor synergism between boldine and doxorubicin, except a possible synergism of i. p. boldine 90 mg/kg combined with doxorubicin when compared with doxorubicin alone.Boldine was cytotoxic to MCF-7 cells and reduced their viability and proliferation in vitro. Exposure to boldine decreased bromodeoxyuridine incorporation and histone H3 phosphorylation but did not induce apoptosis. Boldine treatment resulted in p38, ERK, and JNK activation in the mitogen-activated protein kinase pathway in a dose-dependent manner. Since bioavailability in mice seems to be different from that reported in rats, pharmacokinetic studies in humans are needed to evaluate the role of boldine in the beneficial effects of Boldo infusions.

Comparative study of various subpopulations of cytotoxic cells in blood and ascites from patients with ovarian carcinoma.

AIM OF THE STUDY: A number of observations have indicated that the immune system plays a significant role in patients with epithelial ovarian cancer (EOC). In cases of EOC, the prognostic significance of tumour infiltrating lymphocytes has not been clearly explained yet. The aim is to determine the phenotype and activation molecules of cytotoxic T cell and NK cell subpopulations and to compare their representation in malignant ascites and peripheral blood in patients with ovarian cancer. MATERIAL AND METHODS: Cytotoxic cells taken from blood samples of the cubital vein and malignant ascites were obtained from 53 patients with EOC. Their surface and activation characteristics were determined by means of a flow cytometer. Immunophenotype multiparametric analysis of peripheral blood lymphocytes (PBLs) and tumour infiltrating lymphocytes (TILs) was carried out. RESULTS: CD3(+) T lymphocytes were the main population of TILs (75.9%) and PBLs (70.9%). The number of activating T cells was significantly higher in TILs: CD3(+)/69(+) 6.7% vs. 0.8% (p < 0.001). The representation of (CD3(-)/16(+)56(+)) NK cells in TILs was significantly higher: 11.0% vs. 5.6% (p = 0.041); likewise CD56(bright) and CD-56(bright) from CD56(+) cells were higher in TILs (both p < 0.001). The activation receptor NKG2D was present in 45.1% of TILs vs. 32.3% of PBLs (p = 0.034), but we did not find a significant difference in the numbers of CD56(+)/NKG2D(+) in TILs and PBLs. CONCLUSIONS: These results prove that the characteristics and intensity of anti-tumour responses are different in compared compartments (ascites/PBLs). The knowledge of phenotype and functions of effector cells is the basic precondition for understanding the anti-tumour immune response.

The prognostic significance of tumor epidermal growth factor receptor (EGFR) expression change after neoadjuvant chemoradiation in patients with rectal adenocarcinoma.

AIM OF THE STUDY: The aim of this retrospective study was to determine the prognostic impact of epidermal growth factor receptor (EGFR) expression changes during neoadjuvant chemoradiotherapy in patients with locally advanced rectal cancer. MATERIAL AND METHODS: Fifty patients with locally advanced rectal cancer were evaluated. All the patients were administered the total dose of 44 Gy. Capecitabine has been concomitantly administered in the dose 825 mg/m(2) in two daily oral administrations. Surgery was indicated 4-8 weeks from the chemoradiotherapy completion. Epidermal growth factor receptor expression in the pretreatment biopsies and in the resected specimens was assessed with immunohistochemistry. RESULTS: All of 50 patients received radiotherapy without interruption up to the total planned dose. In 30 patients sphincter-saving surgery was performed, 20 patients underwent amputation of the rectum. Downstaging was described in 30 patients. Four patients have had complete pathologic remission. Twenty-six patients have had partial remission, the disease was stable in 15 patients. Progression was reported in 5 patients. The median disease-free survival was 64.9 months, median overall survival was 76.4 months. Increased EGFR expression was found in 12 patients (26.1%). A statistically significantly shorter overall survival (p < 0.0001) and disease-free survival (p < 0.0001) was found in patients with increased expression of EGFR compared with patients where no increase in the expression of EGFR during neoadjuvant chemoradiotherapy was observed. CONCLUSIONS: The overexpression of EGFR during neoadjuvant chemoradiotherapy for locally advanced rectal adenokarcinoma associated with significant shorter overall survival and disease free survival.