Effect of calcium on the membrane potential ofAmphiuma red cells

Summary An increase in extracellular Ca concentration causes the membrane of giant red cells of the salamander,Amphiuma means, to undergo a marked, transient hyperpolarization. This hyperpolarization is caused by an increase in K permeability of the membrane as judged from the K sensitivity of the membrane potential and from the rate of K loss under influence of raised extracellular Ca concentration. At constant external pH, the induction of hyperpolarization by increased extracellular Ca has a relatively well-defined threshold concentration. Furthermore the phenomenon is of an all or none type with most of the cells having membrane potential values either in the normal range (about –15 mV) or in the range –40 to –70 mV. Shortly after suspension in Ringer's with 15mM Ca, most if not all of the individual cells are hyperpolarized. Upon continued exposure (5–20 min) to the higher Ca concentration the membrane potential returns to the normal value in a fashion compatible with an all or none response. The observed Ca effect is sensitive to the pH of the suspending medium. At pH 6.2 the response is absent whereas the hyperpolarization is markedly stronger at pH 8.2 than at pH 7.2. It is argued that a reliable transport number for K under influence of Ca cannot be estimated from the slope of membrane potentialvs. log (extracellular K concentration). This is probably related to the fact that the membrane potentials of the cells in the population do not stay constant in time. The above phenomenon is compared with the Ca-induced K permeability in poisoned human red cells or red cell ghosts. It is important to note that the cells employed in the present study are neither poisoned nor mechanically disrupted. This study emphasizes that the role of Ca in regulating cell membrane permeability to K seems to be a general feature.     

Measurement of digital blood pressure after local cooling

A double-inlet plastic cuff was designed for local cooling and systolic blood pressure measurement on the middle phalanx of the fingers. With a tourniquet on the proximal phalanx of one finger, cooling for 5 min made the digital artery temperature equal the skin temperature. The difference between the systolic pressure in a control finger and in the cooled finger give the reopening pressure in the digital arteries. At 30, 25, 20, 15, and 10 degrees C, respectively the percent decrease of the finger pressure was 0.2 (0.2), 1.5 (2.5), 8.5 (3.7), 11.4 (3.4), and 15.3 (3.1) in normal young women. In patients with primary or secondary Raynaud's phenomenon, the arterial tone showed an abrupt increase that most often led to complete closure of the digital arteries. The pathological response was expressed as an increased threshold temperature or a well-defined closing temperature that showed only small variations during standardized conditions.     

Stereochemical determination and bioactivity assessment of (S)-(+)-curcuphenol dimers isolated from the marine sponge Didiscus aceratus and synthesized through laccase biocatalysis

Electrospray ionization mass spectrometry-guided isolation of extracts from Didiscus aceratus led to the discovery of several new derivatives of the bioactive bisabolene-type sponge metabolite (S)-(+)-curcuphenol (1). The compounds obtained by this method included a mixture of known (2) and new (3) dihydroxylated analogs as well as a novel family of dimeric derivatives, dicurcuphenols A-E (4-8), and dicurcuphenol ether F (9). Dimers 4-9 were also subsequently obtained through a hemisynthetic method in which 1 was incubated with the enzyme laccase. Atropisomeric dimers 5 and 6 were subjected to vibrational circular dichroism analysis thereby establishing their absolute biaryl axial chirality as P and M, respectively. In contrast to 1, metabolites 2-9 exhibited weak or no cytotoxic or lipoxygenase inhibitory effects.     

Distribution of elements binding to molecules with different molecular weights in aqueous extract of Antarctic krill by size-exclusion chromatography coupled with inductively coupled plasma mass spectrometry

The distribution of silver, arsenic, cadmium, cobalt, chromium, copper, iron, manganese, nickel, lead, selenium and zinc binding to species with different molecular weight in aqueous extract of krill was studied by on-line size-exclusion chromatography (SEC)/inductively coupled plasma mass spectrometry (ICP-MS). The extract was fractionated in three fractions with different molecular weight (MW) ranges (>20,000 relative molecular mass (rel. mol. mass), 2000-20,000 rel. mol. mass and <2000 rel. mol. mass), which were further analyzed by SEC with columns having different optimum fractionation ranges in order to obtain more detailed information about the MW distribution of the elements. Various distribution profiles for the target elements among different MW ranges were observed. The results obtained indicated that manganese, zinc, silver, cadmium and lead species were mostly distributed in the higher MW range (>20,000 rel. mol. mass). In the case of chromium, iron, cobalt, arsenic and selenium, most of them bind to species with lower MW (<2000 rel. mol. mass). Only copper and nickel species was predominantly present in middle MW range (2000-20,000 rel. mol. mass). Further speciation of arsenic compounds in the small MW fraction was carried out with anion exchange chromatography (AEC) coupled with ICP-MS. The results showed that the dominant arsenic species in this fraction is As(III) (63% of extractable arsenic), while As(V) (13%) and two unknown arsenic species (19% and 5%, respectively) are present in lower amounts.     

Mechanisms and function of autophagy in intestinal disease

The discovery of numerous genetic variants in the human genome that are associated with inflammatory bowel disease (IBD) has revealed critical pathways that play important roles in intestinal homeostasis. These genetic studies have identified a critical role for macroautophagy/autophagy and more recently, lysosomal function, in maintaining the intestinal barrier and mucosal homeostasis. This review highlights recent work on the functional characterization of IBD-associated human genetic variants in cell type-specific functions for autophagy.     

Low Completeness of Bacteraemia Registration in the Danish National Patient Registry

Bacteraemia is associated with significant morbidity and mortality and timely access to relia-ble information is essential for health care administrators. Therefore, we investigated the complete-ness of bacteraemia registration in the Danish National Patient Registry (DNPR) containing hospital discharge diagnoses and surgical procedures for all non-psychiatric patients. As gold standard we identified bacteraemia patients in three defined areas of Denmark (~2.3 million inhabitants) from 2000 through 2011 by use of blood culture data retrieved from electronic microbiology databases. Diagnoses coded according to the International Classification of Diseases, version 10, and surgical procedure codes were retrieved from the DNPR. The codes were categorized into seven groups, ranked a priori according to the likelihood of bacteraemia. Completeness was analysed by contin-gency tables, for all patients and subgroups. We identified 58,139 bacteraemic episodes in 48,450 patients; 37,740 episodes (64.9%) were covered by one or more discharge diagnoses within the sev-en diagnosis/surgery groups and 18,786 episodes (32.3%) had a code within the highest priority group. Completeness varied substantially according to speciality (from 17.9% for surgical to 36.4% for medical), place of acquisition (from 26.0% for nosocomial to 36.2% for community), and mi-croorganism (from 19.5% for anaerobic Gram-negative bacteria to 36.8% for haemolytic strepto-cocci). The completeness increased from 25.1% in 2000 to 35.1% in 2011. In conclusion, one third of the bacteraemic episodes did not have a relevant diagnosis in the Danish administrative registry recording all non-psychiatric contacts. This source of information should be used cautiously to iden-tify patients with bacteraemia.     

Action spectra of microalgal photosynthesis and depth distribution of spectral scalar irradiance in a coastal marine sediment of Limfjorden, Denmark

Abstract The role of complementary spectral utilization of light for the zonation of different groups of oxygenic phototrophic organisms in sediments was studied. The marine sediment was covered by a dense population of diatoms with an underlying population of cyanobacteria. Action spectra for photosynthesis and spectral scalar irradiance, E ₀, were measured directly in the sediment at a spatial resolution of 0.1 mm by the use of oxygen and light microsensors. The action spectrum for the diatoms was similar to the attenuation spectrum of the scalar irradiance, K ₀, in the diatom layer with Chl. a and carotenoids being the major photosynthetic pigments. The action spectrum of the cyanobacteria showed photosynthesis maxima at the absorption regions of Chl. a and phycocyanin. The measured depth distribution of spectral scalar irradiance and the action spectra of diatoms and cyanobacteria were used to calculate the spectral quality for photosynthesis of the 400–700 nm light to which the two populations were exposed. This spectral quality was compared to that of the light incident on the sediment surface. Due to preferential extinction of wavelengths, at which their photosynthetically active pigments had maximal absorption, the relative light quality for diatoms was reduced to 85% of the quality of incident light at a similar total quantum flux. This effect was partly due to spectral alterations of light backscattered from the underlying sediment with cyanobacteria. The cyanobacteria at the bottom of the euphotic zone, in contrast, experienced a light spectrum which was favorably altered, to 107% in quality, due to absorption by the overlying diatoms. It was concluded that these changes in spectral light quality can be considered as only one of more factors explaining the zonation of the two phototrophic populations, and that total light intensity and the chemical microenvironment are probably more important factors.     

The Application of New Molecular Methods in the Investigation of a Waterborne Outbreak of Norovirus in Denmark, 2012

In December 2012, an outbreak of acute gastrointestinal illness occurred in a geographical distinct area in Denmark covering 368 households. A combined microbiological, epidemiological and environmental investigation was initiated to understand the outbreak magnitude, pathogen(s) and vehicle in order to control the outbreak. Norovirus GII.4 New Orleans 2009 variant was detected in 15 of 17 individual stool samples from 14 households. Norovirus genomic material from water samples was detected and quantified and sequencing of longer parts of the viral capsid region (>1000 nt) were applied to patient and water samples. All five purposely selected water samples tested positive for norovirus GII in levels up to 1.8×10⁴ genomic units per 200 ml. Identical norovirus sequences were found in all 5 sequenced stool samples and 1 sequenced water sample, a second sequenced water sample showed 1 nt (<0.1%) difference. In a cohort study, including 256 participants, cases were defined as residents of the area experiencing diarrhoea or vomiting onset on 12–14 December 2012. We found an attack rate of 51%. Being a case was associated with drinking tap-water on 12–13 December (relative risk = 6.0, 95%CI: 1.6–22) and a dose-response relation for the mean glasses of tap-water consumed was observed. Environmental investigations suggested contamination from a sewage pipe to the drinking water due to fall in pressure during water supply system renovations. The combined microbiological, epidemiological and environmental investigations strongly indicates the outbreak was caused by norovirus contamination of the water supply system.