Survivin antiapoptotic gene expression as a prognostic factor in non-small cell lung cancer: in situ hybridization study.

Survivin is an inhibitor of apoptosis that plays a significant role in cell cycle regulation and is important for survival prognosis in many neoplasms. Survivin expression was assessed by in situ hybridization (ISH) in 60 consecutive patients (54 males and 4 females) with NSCLC treated between 1993 and 1997. The examined patients had IIB and IIIA stage according to TNM system. In all cases the chemotherapy with cisplatin and etoposide (2 cycles) was administered prior the surgery; in patients responding to the therapy one more cycle was applied. Survivin gene overexpression was observed in 35 patients (58.3%). There was no correlation between survivin mRNA level and histological type of tumor, stage of cell differentiation, stage of disease according to TNM classification, performance status according to WHO and number of chemotherapy regimens administered (p > 0.05). However, the correlation between survivin gene expression and response to the chemotherapy was statistically significant (p = 0.04). Statistical analysis showed that median survival in patients with survivin gene overexpression was shorter (14.0 months) as compared to patients with no expression (60.0 months; p = 0.00002). In survival assessment by means of Kaplan-Meier test, 14.3% of five-year survival was achieved in the former group versus 60% in the latter (p = 0.00003). Univariate analysis (log-rank test) showed that significant independent prognostic factors in NSCLC included: stage of the disease according to TNM classification (p = 0.006), response to chemotherapy (p = 0.005) and pattern of survivin gene expression (p = 0.00003). Multivariate analysis utilizing Cox's model showed that for survival assessment the stage according to TNM, response to the chemotherapy and survivin expression estimated by means of ISH are of statistical significance (p=0.00001). The calculated predictive values showed that ISH technique was quite accurate in assessment of five-year survival. Our data show that survivin expression may be used as a prognostic factor and a target for therapy

Chicken intestinal organoids: a novel method to measure the mode of action of feed additives

There is a rapidly growing interest in how the avian intestine is affected by dietary components and feed additives. The paucity of physiologically relevant models has limited research in this field of poultry gut health and led to an over-reliance on the use of live birds for experiments. The development of complex 3D intestinal organoids or "mini-guts" has created ample opportunities for poultry research in this field. A major advantage of the floating chicken intestinal organoids is the combination of a complex cell system with an easily accessible apical-out orientation grown in a simple culture medium without an extracellular matrix. The objective was to investigate the impact of a commercial proprietary blend of organic acids and essential oils (OA+EO) on the innate immune responses and kinome of chicken intestinal organoids in a Salmonella challenge model. To mimic the in vivo prolonged exposure of the intestine to the product, the intestinal organoids were treated for 2 days with 0.5 or 0.25 mg/mL OA+EO and either uninfected or infected with Salmonella and bacterial load in the organoids was quantified at 3 hours post infection. The bacteria were also treated with OA+EO for 1 day prior to challenge of the organoids to mimic intestinal exposure. The treatment of the organoids with OA+EO resulted in a significant decrease in the bacterial load compared to untreated infected organoids. The expression of 88 innate immune genes was investigated using a high throughput qPCR array, measuring the expression of 88 innate immune genes. Salmonella invasion of the untreated intestinal organoids resulted in a significant increase in the expression of inflammatory cytokine and chemokines as well as genes involved in intracellular signalling. In contrast, when the organoids were treated with OA+EO and challenged with Salmonella, the inflammatory responses were significantly downregulated. The kinome array data suggested decreased phosphorylation elicited by the OA+EO with Salmonella in agreement with the gene expression data sets. This study demonstrates that the in vitro chicken intestinal organoids are a new tool to measure the effect of the feed additives in a bacterial challenge model by measuring innate immune and protein kinases responses

Maseni udjeli polifenola, antocijanina i resveratrola u kultivarima brusnice te njihova antioksidacijska svojstva

The study involved an evaluation of five cranberry cultivars grown in Poland: Ben Lear, Pilgrim, Stevens, Early Richard and Bergman. The reference sample comprised wild-grown common cranberry (Vaccinium oxycoccus). The mass fractions of total phenolic compounds, anthocyanins and resveratrol (HPLC-DAD), as well as the antioxidant properties (DPPH·, ·OH and ABTS+ radical scavenging capacity) were determined. Statistically significant differences (p<0.05) were reported as regards the mass fractions of polyphenols and anthocyanins in the fruit of the analysed cultivars. The highest polyphenol mass fraction was determined in Ben Lear (374.2 mg per 100 g of fresh mass), while Early Richard was the richest source of anthocyanins (77.1 mg per 100 g of fresh mass). The fruit of common cranberry contained the highest quantities of resveratrol (712.3 ng/g of fresh mass), and its mass fraction in the investigated cultivars ranged from 533.4 (cv. Stevens) to 598.2 ng/g of fresh mass (Ben Lear). Common cranberry was also marked by the highest ABTS+ scavenging capacity. Stevens and Pilgrim were characterised by a strong capability to scavenge DPPH· and ·OH free radicals. Statistically significant differences (p<0.05) were observed in respect of the free radical scavenging capacity of most investigated cranberry cultivars.Ispitano je pet kultivara brusnica koji se uzgajaju u Poljskoj: Ben Lear, Pilgrim, Stevens, Early Richard i Bergman. Referentni je uzorak bila divlja brusnica (Vaccinium oxycoccus). Određeni su maseni udjeli ukupnih fenola, antocijanina i resveratrola (pomoću HPLC-DAD metode), te antioksidacijska svojstva brusnica (tj. sposobnost uklanjanja DPPH˙, OH˙ i ABTS+ radikala). Utvrđene su statistički značajne razlike (p<0,05) između masenih udjela polifenola i antocijanina u plodovima ispitanih kultivara. Najveći maseni udio polifenola ima kultivar Ben Lear (374,2 mg/100 g svježih plodova), dok su plodovi kultivara Early Richard najbogatiji antocijaninima (77,1 mg/100 g svježih plodova). Maseni udio resveratrola u plodovima kultivara varira od 533,4 (u kultivaru Stevens) do 598,2 ng/g svježih plodova (u kultivaru Ben Lear). Plodovi divlje brusnice sadrže najviše resveratrola (712,3 ng/g svježih plodova). Divlja je brusnica također pokazala najbolju sposobnost uklanjanja ABTS+ radikala, dok kultivari Stevens i Pilgrim imaju znatnu moć uklanjanja DPPH˙ i OH˙ radikala. Utvrđene su statistički značajne razlike (p<0,05) u sposobnosti uklanjanja slobodnih radikla između većine kultivara brusnice

Surgery followed by irradiation in glioblastoma multiforme. A report of 28 cases

Treatment results in glioblastoma multiforme, irrespective of the management, are poor. Median survival in patients managed with surgery alone is 4 months and in those treated with surgery and adjuvant radiotherpy -9 months.Twenty eight patients with glioblastoma multiforme were treated at the Department of Oncology and Radiotherapy, Medical University of Gdańsk between 1991 to 1995. There were females and 20 males and the median age was 58 years (range 18 to 75 years). In 22 cases (78%) diagnosis was confirmed by histology, and in the remaining six cases biopsy was not taken due to the deep localization of the tumour; in all these patients diagnosis was based on CT imaging. All patients were irradiated with cobalt unit and received conventional radiotherapy, 5 days a week, 1.8 Gy per fraction. The first part of treatment included whole brain irradiation (40 Gy) delivered through lateral parallel opposed fields. Thereafter in all instances a brain CT was done and in case of regression or stabilisation (23 pts), a boost dose of 15–22 Gy with reduced portals was delivered. Total dose delivered to the tumor bed was 55–62 Gy. Radiotherapy tolerance was satisfactory and there were no serious complications and interruptions of treatment.Median local recurrence-free survival was 5.3 months, and a median survival – 9.9 months (range, 1.6 to 31.2 months). There was no correlation between survival and radiotherapy dose, sex, pretreatment WHO performance status and tumor localization.Our results confirm poor prognosis in glioblastoma multitorme. New more effective therapeutic approaches are sorely needed in this tumour

Inference in Cardiovascular Modelling Subject to Medical Interventions

Pulmonary hypertension (PH), i.e., high blood pressure in the lungs, is a serious medical condition that can damage the right ventricle of the heart and ultimately lead to heart failure. Standard diagnostic procedures are based on right-heart catheterization, which is an invasive technique that can potentially have serious side effects. Recent methodological advancements in fluid dynamics modelling of the pulmonary blood circulation system promise to mathematically predict the blood pressure based on non-invasive measurements of the blood flow. Thus, subsequent to PH diagnostication, further investigations would no longer require catheterization. However, in order for these alternative techniques to be applicable in the clinic, accurate model calibration and parameter estimation are paramount. Medical interventions taken to combat high blood pressure (as predicted from the mathematical model) alter the underlying cardiovascular physiology, thus interfering with the parameter estimation procedure. In the present study, we have carried out a series of cardiovascular simulations to assess the reliability of cardiovascular physiological parameter estimation in the presence of medical interventions. Our principal result is that if the closed-loop effect of medical interventions is accounted for, the model calibration provides accurate parameter estimates. This finding has important implications for the applicability of cardio-physiological modelling in the clinical practice

Elimination of micropollutants in activated sludge reactors with a special focus on the effect of biomass concentration

This study aimed to investigate the effects of sludge retention time (SRT), hydraulic retention time (HRT), and biomass concentration (CTSS) in activated sludge systems on removal of various micropollutants (MPs), covering a wide spectrum of biodegradability. The influence of biomass concentration on the classical pseudo-first-order rate constant was verified. Results showed that the removal rate constants were affected by both the HRT and SRT. The enhancement of the SRT increased the removal of all the MPs except for two macrolide antibiotics. Application of a higher HRT also improved MP removal, as was expected from the measured removal rate constants. More interesting, our results indicated that, logically, the increase of biomass concentration (expressed as total suspended solids CTSS) from 3 to 5 gTSS L−1 significantly enhanced the removal rate of the highly and moderately degradable compounds. Conversely, a further increase to 8 gTSS L−1 produced only an unexpected moderate effect, showing that the rate was not proportional to biomass concentration, contrary to what is generally postulated. Therefore, the use of classical kinetic models is questionable, since they do not cover the entire range of boundary conditions in activated sludge systems. This work opens new research paths and suggests potential improvements to processe

A versatile panel of reference gene assays for the measurement of chicken mRNA by quantitative PCR

Quantitative real-time PCR assays are widely used for the quantification of mRNA within avian experimental samples. Multiple stably-expressed reference genes, selected for the lowest variation in representative samples, can be used to control random technical variation. Reference gene assays must be reliable, have high amplification specificity and efficiency, and not produce signals from contaminating DNA. Whilst recent research papers identify specific genes that are stable in particular tissues and experimental treatments, here we describe a panel of ten avian gene primer and probe sets that can be used to identify suitable reference genes in many experimental contexts. The panel was tested with TaqMan and SYBR Green systems in two experimental scenarios: a tissue collection and virus infection of cultured fibroblasts. GeNorm and NormFinder algorithms were able to select appropriate reference gene sets in each case. We show the effects of using the selected genes on the detection of statistically significant differences in expression. The results are compared with those obtained using 28s ribosomal RNA, the present most widely accepted reference gene in chicken work, identifying circumstances where its use might provide misleading results. Methods for eliminating DNA contamination of RNA reduced, but did not completely remove, detectable DNA. We therefore attached special importance to testing each qPCR assay for absence of signal using DNA template. The assays and analyses developed here provide a useful resource for selecting reference genes for investigations of avian biology