Photodynamic effects of protoporphyrin on the cellular level—Anin vitro approach

Summary The purpose of this study was characterizing the phototoxic action of protoporphyrin and cellular protection mechanisms, as studied on the cellular level. In this process, active oxygen is involved. As a biological system, rat hepatocyte shortterm and primary cultures were used. Phototoxicity of protoporphyrin could be observed, after previous absorption of protoporphyrin to membrane structures. Damaging of several cell organelles occurred, such as mitochondria and lysosomes. Peroxisomes were not affected. Coated vesicles located at the periphery of the cells’ interior suggested that protoporphyrin absorption is mediated by an active uptake (endocytosis), as well as passive diffusion. Lipid peroxidation played a role in protoporphyrin photoxicity. Cellular protection mechanisms such as superoxide dismutase and the scavenger glutathione (GSH) protected the cells from active oxygen toxicity. In conclusion, protoporphyrin entered the cells by diffusion and endocytosis. Previous adsorption to the membrane structures was necessary for the expression of protoporphyrin phototoxicity. However, active oxygen itself could not be demonstrated. Lipid peroxidation was involved in cell-damaging processes. Mechanisms of protoporphyrin phototoxicity on the cellular level were studied. Rat hepatocyte primary and short-term cultures proved to be suitablein vitro systems for studying biochemical and morphological effects on the cellular level. This article is based on PhD research carried out at the Department of Veterinary Pharmacology, Pharmacy and Toxicology, Utrecht University, The Netherlands.     

Objectively Measured Total and Occupational Sedentary Time in Three Work Settings

Background

Sedentary behaviour increases the risk for morbidity. Our primary aim is to determine the proportion and factors associated with objectively measured total and occupational sedentary time in three work settings. Secondary aim is to study the proportion of physical activity and prolonged sedentary bouts.

Methods

Data were obtained using ActiGraph accelerometers from employees of: 1) a financial service provider (n = 49 men, 31 women), 2) two research institutes (n = 30 men, 57 women), and 3) a construction company (n = 38 men). Total (over the whole day) and occupational sedentary time, physical activity and prolonged sedentary bouts (lasting ≥30 minutes) were calculated by work setting. Linear regression analyses were performed to examine general, health and work-related factors associated with sedentary time.

Results

The employees of the financial service provider and the research institutes spent 76–80% of their occupational time in sedentary behaviour, 18–20% in light intensity physical activity and 3–5% in moderate-to-vigorous intensity physical activity. Occupational time in prolonged sedentary bouts was 27–30%. Total time was less sedentary (64–70%), and had more light intensity physical activity (26–33%). The employees of the construction company spent 44% of their occupational time in sedentary behaviour, 49% in light, and 7% in moderate intensity physical activity, and spent 7% in sedentary bouts. Total time spent in sedentary behavior was 56%, 40% in light, and 4% in moderate intensity physical behaviour, and 12% in sedentary bouts. For women, low to intermediate education was the only factor that was negatively associated with occupational sedentary time.

Conclusions

Sedentary behaviour is high among white-collar employees, especially in highly educated women. A relatively small proportion of sedentary time was accrued in sedentary bouts. It is recommended that worksite health promotion efforts should focus on reducing sedentary behaviour through improving light intensity physical activity.     

The 'normalization' of germ-free guineapigs with host-specific caecal microflora

Hysterectomy-derived germ-free guineapigs were given colonization-resistant caecal flora from mice (mCRF) or microflora obtained from the caecum of an antibiotic-decontaminated conventional guineapig (gpCRF) and compared with guineapigs raised conventionally with the sow. Body weight and the following intestinal parameters were determined for the groups: colonization resistance (CR) to Escherichia coli, relative caecal weight (RCW), beta-aspartylglycine (faeces), volatile fatty acids (caecum) and bile acids (faeces). mCRF guineapigs showed values quite different from control animals for CR and RCW, indicating the unsuitability of mouse CRF for normalizing guineapigs. In gpCRF guineapigs CR and RCW values were comparable with controls, indicating the suitability of the guineapig flora for normalizing guineapigs. mCRF guineapigs housed with gpCRF guineapigs, showed an improvement in CR and RCW, yielding values found in control animals.     

KRAS codon 12 mutations occur very frequently in pancreatic adenocarcinomas.

DNAs from human pancreatic adenocarcinomas were analyzed for the presence of mutations in codons 12, 13 and 61 of the NRAS, KRAS and HRAS gene. Formalin-fixed and paraffin-embedded tissue was used directly in an in vitro amplification reaction to expand the relevant RAS sequences. The mutations were detected by selective hybridization using mutation-specific synthetic oligonucleotides. In 28 of the 30 patients we found a mutation in codon 12 of the KRAS gene. This result confirms the findings of Almoguera et al. [Cell 53 (1988) 549-554] that KRAS mutations occur frequently in adenocarcinomas of the exocrine pancreas. The mutations are predominantly G-T transversions, in contrast to the KRAS mutations in colon tumors which are mainly G-A transitions. Furthermore, in a portion of the tumors the mutation appears to be homozygous.     

Relation of out of hours activity by general practice and accident and emergency services with deprivation in Nottingham: longitudinal survey.

OBJECTIVES: To investigate the relation between out of hours activity of general practice and accident and emergency services with deprivation and distance from accident and emergency department. DESIGN: Six month longitudinal study. SETTING: Six general practices and the sole accident and emergency department in Nottingham. SUBJECTS: 4745 out of hours contacts generated by 45,182 patients from 23 electoral wards registered with six practices. MAIN OUTCOME MEASURES: Rates of out of hours contacts for general practice and accident and emergency services calculated by electoral ward; Jarman and Townsend deprivation scores and distance from accident and emergency department of electoral wards. RESULTS: Distances of wards from accident and emergency department ranged from 0.8 to 9 km, and Jarman deprivation scores ranged from -23.4 to 51.8. Out of hours contacts varied by ward from 110 to 350 events/1000 patients/year, and 58% of this variation was explained by the Jarman score. General practice and accident and emergency rates were positively correlated (Pearson coefficient 0.50, P = 0.015). Proximity to accident and emergency department was not significantly associated with increased activity when deprivation was included in regression analysis. One practice had substantially higher out of hours activity (B coefficient 124 (95% confidence interval 67 to 181)) even when deprivation was included in regression analysis. CONCLUSIONS: A disproportionate amount of out of hours workload fell on deprived inner city practices. High general practice and high accident and emergency activity occurred in the same areas rather than one service substituting for the other.     

Evaluation of the prevalence and economic burden of adverse drug reactions presenting to the medical emergency department of a tertiary referral centre: a prospective study

Background  

Adverse drug reactions (ADRs) are now recognized as an important cause of hospital admissions, with a proportion ranging from 0.9–7.9%. They also constitute a significant economic burden. We thus aimed at determining the prevalence and the economic burden of ADRs presenting to Medical Emergency Department (ED) of a tertiary referral center in India     

ENOD40 affects elongation growth in tobacco Bright Yellow-2 cells by alteration of ethylene biosynthesis kinetics

Plant developmental processes are controlled by co-ordinated action of phytohormones and plant genes encoding components of developmental response pathways. ENOD40 was identified as a candidate for such a plant factor with a regulatory role during nodulation. Although its mode of action is poorly understood, several lines of evidence suggest interaction with phytohormone response pathways. This hypothesis was investigated by analysing cytokinin-, auxin-, and ethylene-induced responses on cell growth and cell division in transgenic 35S:NtENOD40 Bright Yellow-2 (BY-2) tobacco cell suspensions. It was found that cell division frequency is controlled by the balance between cytokinin and auxin in wild-type cells and that this regulation is not affected in 35S:NtENOD40 lines. Elongation growth, on the other hand, is reduced upon overexpression of NtENOD40. Analysis of ethylene homeostasis shows that ethylene accumulation is accelerated in 35S:NtENOD40 lines. ENOD40 action can be counteracted by an ethylene perception blocker, indicating that ethylene is a negative regulator of elongation growth in 35S:NtENOD40 cells, and that the NtENOD40-induced response is mediated by alteration of ethylene biosynthesis kinetics.     

N-Alkyl-N-arylmethylpiperidin-4-amines: novel dual inhibitors of serotonin and norepinephrine reuptake

A series of N-alkyl-N-arylmethylpiperidin-4-amines have been prepared and are demonstrated to be inhibitors of both serotonin and norepinephrine reuptake.