Correlation of open cell-attached and excised patch clamp techniques

The excised patch clamp configuration provides a unique technique for some types of single channel analyses, but maintenance of stable, long-lasting preparations may be confounded by rundown and/or rapid loss of seal. Studies were performed on the amiloride-sensitive Na⁺ channel, located on the apical surface of A₆ cells, to determine whether the nystatininduced open cell-attached patch could serve as an alternative configuration.Compared to excised inside-out patches, stable preparations were achieved more readily with the open cell-attached patch (9% vs. 56% of attempts). In both preparations, the current voltage (I-V) relation was linear, current amplitudes were equal at opposite equivalent clamped voltages, and E _rev was zero in symmetrical Na⁺ solutions, indicating similar Na⁺ activities on the cytosolic and external surfaces of the patch. Moreover, there was no evidence that nystatin altered channel activity in the patch because slope conductance (3–4pS) and E _rev (75 mV), when the bath was perfused with a high K:low Na solution (E _Na=80 mV), were nearly equal in both patch configurations.Our results therefore indicate that the nystatininduced open cell-attached patch can serve as an alternative approach to the excised inside-out patch when experiments require modulation of univalent ions in the cytosol.We thank Dr. Olaf S. Andersen for his suggestions in the development of the open cell-attached recording technique. This work was supported by a National Institutes of Health grant (DK-18061)     

Autofluorescence imaging for recurrence detection in skin cancer postoperative scars

This clinical study is a first attempt to use autofluorescence for recurrence diagnosis of skin cancer in postoperative scars. The proposed diagnostic parameter is based on a reduction in scar autofluorescence, evaluated in the green spectral channel. The validity of the method has been tested on 110 postoperative scars from 56 patients suspected of non‐melanoma skin cancer, with eight patients (13 scars) available for the repeated examination. The recurrence diagnosis within a scar has been made after two subsequent autofluorescence check‐ups, representing the temporal difference between the scar autofluorescence amplitudes as a vector. The recognition of recurrence has been discussed to represent the significant deviations from the value of vector angle θ. This new autofluorescence‐based method can be easily integrated into the postoperative monitoring of surgical scars and can help diagnose the recurrence of skin cancer from the early stage of scar development.     

The 5′ untranslated region of Arabidopsis thaliana calmodulin cDNA is an independent cDNA containing an open reading frame

A cDNA clone, pAUK1, with an open reading frame (ORF) coding for a hypothetical 164-amino-acid protein was isolated from an Arabidopsis thaliana (L.) Heynh cDNA library. The clone was attached, tail to tail, to the 3′ end of A. thaliana hexokinase cDNA. An almost identical sequence had been previously described as the 5′ untranslated region (5′ UTR) of A. thaliana calmodulin cDNA (ACaM-2). Sequence comparison with three additional A. thaliana truncated cDNA clones which appear in a database (GenBank) supports the conclusion that pAUKl is identical to the 5′ UTR of ACaM-2 and that the 5′ UTR of ACaM-2 is an independent cDNA artificially linked to A. thaliana calmodulin cDNA.     

Le Jas d’en Biel 2, nouveau site gravettien du piémont est-méditerranéen des Pyrénées

The Corbières massif forms the lowest part of the Eastern Pyrenees, and as such is the easiest natural passage to connect the coastal plains of Languedoc and Roussillon in Southern France with those of Gironès and Barcelona in Northern Spanish Catalonia. This geographical situation confers on each of the sites discovered in this passage a role of milestone that can allow us to apprehend the dynamics of anthropic movements and relationships across these spaces. In 2007, the presence of a first Gravettian site in this Northern Pyrenean foothill was published: Jas d’en Biel 1. This discovery filled an “archaeological void” between the Gravettian sites of Languedoc and those of Spanish Catalonia. The discovery of a new site: Jas d’en Biel 2, 300 m from the first, reinforces the Gravettian presence in this region. The fact that the two sites present similar characteristics in terms of anthropic choices (solar orientation, proximity to a watercourse, lightness of the soil, protection from the wind, etc.) are all parameters that demonstrate the choices made by Gravettian men. The respective compositions of the lithic industries of the series do not show significant differences, allowing us to imagine a certain synchronism between the two sites. Burins are the most numerous tools, but at Jas d’en Biel 2 we find specimens close to the Raysse type that were absent from Jas d’en Biel 1. Jas d’en Biel 2, as well as its neighbor Jas d’en Biel 1, shows similarities with certain sites in Languedoc, such as Bois-des-Brousses, La Treille, La Verrière, the caves of Bize or La Crouzade. These are usually small deposits that belong to terminal moments of the Gravettian period. All these parameters point to similarities between the Languedoc sites and Jas d’en Biel 1 and 2. The geographical position of the Corbières massif allows us to consider the Jas d’en Biel sites as the last milestones of the Gravettian period upstream from the crossing of the Pyrenean passes, before the sites of the Serinyà region, such as l’Arbreda. Recent surveys carried out in the same valley of the Ravin d’en Saman have made it possible to locate and identify several other open-air settlements related to the Gravettian lato sensu. An analysis of intra- and extra-site characteristics will aim to understand the parameters that led men to settle in this micro-zone, as well as the reasons for this “concentration”.     

Sense in antisense?

A correspondence between open reading frames in sense and antisense strands is expected from the hypothesis that the prototypic triplet code was of general form RNY, where R is a purine base, N is any base, and Y is a pyrimidine. A deficit of stop codons in the antisense strand (and thus long open reading frames) is predicted for organisms with high G + C percentages; however, two bacteria (Azotobacter vinelandii, Rhodobacter capsulatum) have larger average antisense strand open reading frames than predicted from (G + C)%. The similar Codon frequencies found in sense and antisense strands can be attributed to the wide distribution of inverted repeats (stem-loop potential) in natural DNA sequences.     

The response of plasma gastric-inhibitory polypeptide (GIP) to slow and fast glucose ingestion in Billroth II resected patients and normal controls

Eight Billroth II resected patients and 8 normal controls were given two oral glucose loads, one ingested within 2 min, and the other ingested slowly over 80 min. In the Billroth II resected group, the integrated plasma GIP release was significantly higher after the fast than after the slow glucose ingestion. In this group the integrated plasma GIP release was also significantly higher than in the control group, but only after the fast glucose ingestion. These findings indicate that the rate of glucose delivery into the intestine may be of importance in the plasma GIP response to oral glucose.     

Secretin modulation of behavioral and physiological functions in the rat

The effect of secretin on behavioral and physiological functions in the rat was investigated. Secretin injected intracerebroventricularly (ICV) significantly increased defecation and decreased novel-object approaches in rats. The peptide showed no significant effects on stereotypic behavior (gnawing, grooming and rearing), open-field locomotor activity however was significantly decreased, an effect that was probably due to a decreased propensity for the rats to initiate locomotor responses. In addition, secretin showed significant effects on respiration rate in anesthetized rats. When the peptide was injected in the lateral ventricle a decrease in respiration rate occurred, but when the brain was perfused from the lateral ventricle to the cisterna magna increases in respiration rate occurred. These data, combined with the facts that secretin and secretin receptors have been identified in the brain indicate that secretin may play a neurotransmitter or neuroregulator role in the central nervous system.     

Fine structural distribution of the surface-connected canalicular system in frog thrombocytes

Summary The existence of the surface-connected canalicular system (SCCS) has been demonstrated in semithick sections of the frog thrombocytes by the use of a high voltage electron microscope. The SCCS of the thrombocytes in Rana catesbeiana and Rana nigromaculata consists of numerous canaliculi and vesicles with a diameter of 250 nm, which join with one another to make a complex network throughout the cytoplasm. Although the SCCS of Xenopus laevis fits well into the pattern described in Rana catesbeiana, the diameter of the canaliculi of the SCCS is about 500 nm. The results of this study suggest that the SCCS is a specific organelle of the thrombocyte system common to submammals and mammals.     

Breakup and re-formation of colony in the first-instar larvae of the winter cherry bug,Acanthocoris sordidus thunberg (Hemiptera: Coreidae), in relation to the defence against their enemies

The winter cherry bug, Acanthocoris sordidusThunberg , lives in aggregation especially in their early larval instars. Using the 1st-instar larvae of this species, the author tried to clarify both the processes and the mechanisms of the breakup and later re-formation of colony in relation to the defence against their enemies. The results obtained were summarized as follows.

1. In the field population, there is a high possibility of dispersal of the 1st-instar larvae from a colony possibly through the disturbance by some predators but they can re-form a colony with each other or join, with colonies of different instar larvae.
2. The individuals in a colony immediately disperse through the attack of predatory coccinellid beetle, Harmonica axyridis but tend to re-form a colony in a short time.
3. The breakup of colony is caused by the secretion from the attacked individual.
4. The formation of colony is attributed to the habit closely related with the senses of smell and/or contact.

From these results, it was concluded that the dispersal of 1st-instar larvae from a colony, followed by the re-formation of a colony, is an an adaptive behaviour to escape from the attack by their predators.     

Specific features of ex‐obese patients significantly influence the functional cell properties of adipose‐derived stromal cells

Adipose‐derived stromal cells (ADSC) are increasingly used in clinical applications due to their regenerative capabilities. However, ADSC therapies show variable results. This study analysed the effects of specific factors of ex‐obese patients on ADSC functions. ADSC were harvested from abdominal tissues (N = 20) after massive weight loss. Patients were grouped according to age, sex, current and maximum body mass index (BMI), BMI difference, weight loss method, smoking and infection at the surgical site. ADSC surface markers, viability, migration, transmigration, sprouting, differentiation potential, cytokine secretion, telomere length and mtDNA copy number were analysed. All ADSC expressed CD73, CD90, CD105, while functional properties differed significantly among patients. A high BMI difference due to massive weight loss was negatively correlated with ADSC proliferation, migration and transmigration, while age, sex or weight loss method had a smaller effect. ADSC from female and younger donors and individuals after weight loss by increase of exercise and diet change had a higher activity. Telomere length, mtDNA copy number, differentiation potential and the secretome did not correlate with patient factors or cell function. Therefore, we suggest that factors such as age, sex, increase of exercise and especially weight loss should be considered for patient selection and planning of regenerative therapies.