The genetic basis of a craniofacial disease provides insight into COPII coat assembly

Proteins trafficking through the secretory pathway must first exit the endoplasmic reticulum (ER) through membrane vesicles created and regulated by the COPII coat protein complex. Cranio-lenticulo-sutural dysplasia (CLSD) was recently shown to be caused by a missense mutation in SEC23A, a gene encoding one of two paralogous COPII coat proteins. We now elucidate the molecular mechanism underlying this disease. In vitro assays reveal that the mutant form of SEC23A poorly recruits the Sec13-Sec31 complex, inhibiting vesicle formation. Surprisingly, this effect is modulated by the Sar1 GTPase paralog used in the reaction, indicating distinct affinities of the two human Sar1 paralogs for the Sec13-Sec31 complex. Patient cells accumulate numerous tubular cargo-containing ER exit sites devoid of observable membrane coat, likely representing an intermediate step in COPII vesicle formation. Our results indicate that the Sar1-Sec23-Sec24 prebudding complex is sufficient to form cargo-containing tubules in vivo, whereas the Sec13-Sec31 complex is required for membrane fission.     

Modification of redox equilibria between heme and flavin within yeast flavocytochrome b2 (l-lactate cytochrome c reductase) upon binding of pyruvate,the reaction product

Direct determinations of the concentration of semiquinone spin in redox equilibrium with the cytochrome b₂ moiety were carried out at room temperature in the presence of added pyruvate or in its absence. Results show that redox potentials of the one-electron couples of the prosthetic flavin are markedly affected by binding of pyruvate the reaction product in the oxidation of l-lactate. The proportion of flavin semiquinone nearly reaches then 100 per cent.     

What counts? Volunteers and their organisations in the recording and monitoring of biodiversity

There is a pressing need for volunteer amateur naturalists to participate in data collection for biodiversity monitoring programmes in Europe. It is being addressed in some countries, but less so in others. This paper discusses the results from qualitative research using semi-structured interviews, focus groups and participant observation within nine Participatory Monitoring Network (PMN) organisations in six European countries. The paper examines the features that facilitate recruitment, retention and motivations of volunteers to participate in biodiversity monitoring, including the social and cultural milieus in which they operate. The paper concludes that volunteers place a high degree of significance on their social experience within PMNs. Successful creation and management of PMNs thus requires that similar levels of attention be paid to social aspects of the organisation as are paid to the generation and management of data.     

Characterization of the sensitivity of side scatter in a flow-stream waveguide flow cytometer.

BACKGROUND: We previously reported a new optical configuration, in which both the side scatter and the fluorescence are collected using the index-guided, total internal reflection of a flow stream in air (the flow-stream waveguide). METHODS: Using a mixture of 0.202-microm and 0.093-microm diameter polystyrene beads, we have characterized the side scatter (SSC) sensitivity of a custom-built flow cytometer (miniFlo) which incorporates a flow-stream waveguide. RESULTS: The SSC-triggered SSC signal of 0.093-microm polystyrene beads in water was almost baseline resolved from the background. We also measured the SSC-triggered SSC signal of the same beads in water on our FACScan, which is a commercial unit with the conventional optical arrangement that uses a custom imaging objective to collect light from a sheath flow cuvette in perpendicular direction-the signal from 0.093-microm beads was not resolved from the background. CONCLUSIONS: The SSC sensitivity of miniFlo is one of the best reported in the literature. Cytometry 37:160-163, 1999. Published 1999 Wiley-Liss, Inc.     

Aerobiological monitoring for fungal spores in a rehabilitation hospital in Northern Italy

In the setting of an aerobiological study of confined environments and environmental prevention of nosocomial infections, aerobiological monitoring of 22 areas in Montescano Medical Rehabilitation Centre (Pavia) was performed in three different months: February, May, and August, 1994. A Lanzoni 3000 spore trap was used for the monitoring. Examination of the results showed that, in the environments monitored, the indoor presence of fungi increased progresively over the three periods considered. The behaviour of individual genera of fungi differed. In the winter periodPenicillium andAspergillus were the most commonly encountered genera,Cladosporium was the dominant genus from the spring to the summer while the yeasts had their maximum counts in the spring morning. The use of air conditioning, combined with the low air exchange between outdoors and andoors in some of the environments certainly played a part in the low concentrations of these latter aerofungi.     

Venous blood lactate evaluation in equine neonatal intensive care

The use of blood lactate concentration as an indicator of prognosis and disease severity has become a common practice in equine medicine, especially with the validation of handheld analyzers. However, few authors described lactate concentration in critically ill foals, and there are no published studies about the use of handheld analyzers in neonatal foals. In this study, for the first time in the equine neonate, we validated the Lactate Scout analyzer, both in healthy and in critically ill foals. The study also describes the normal range for blood lactate in 26 healthy neonatal foals during the first 72 h of life. Moreover, the utility of venous lactate measurement in 88 critically ill foals was determined, describing lactate values in the most common neonatal pathologies, evaluating serial blood lactate measurements, and investigating its prognostic value. The comparison with the enzymatic-colorimetric reference method showed that the Lactate Scout analyzer is reliable. The mean difference (bias ±2SD) between the two methods was close to zero for all comparisons, and the SD of difference was ±0.76 with a 95% confidence interval from −1.58 to 1.40 mmol/L. In healthy foals, blood lactate concentrations at birth and at 12 h of life were statistically higher (P < 0.01) than lactate concentrations measured at subsequent times. In critically ill foals, the highest lactate concentration at admission was found in hemorrhagic shock, septic shock, and complicated perinatal asphyxia syndrome (PAS). Our results showed that hyperlactatemia, although it does not provide diagnostic information, indicates the severity of illness and the need for an early and aggressive intervention. This could be very useful both during hospitalization and in the field to support veterinarians in making a decision about referral. Furthermore lactatemia proved to be a reliable prognostic parameter: In nonsurviving foals, hyperlactatemia persisted during the entire hospitalization, whereas in survivors there were no significant differences after 24 h from admission. Because prognostic parameters have certain limitations, hyperlactatemia should not be used alone to decide whether to discontinue treatments in critically ill foals. A careful and complete clinical examination is always essential.     

Enhancing effect of lithium and potassium ions on lectin-induced lymphocyte proliferation in aging and down's syndrome subjects

The effect of different concentrations of LiCl or KCl (0.6–20 meq/liter) on PHA-stimulated lymphocytes from young, old, and Down's syndrome subjects was studied. LiCl showed a dramatic enhancing effect on [³H]thymidine incorporation induced by a suboptimal dose of PHA in old subjects and Down's syndrome patients. An increase of [³H]thymidine incorporation in human lymphocytes stimulated by a suboptimal dose of PHA was also observed with KCl. This effect was higher in old subjects than that observed in young and Down's subjects. LiCl and KCl can modulate and partially restore the derangement in early events of mitogen stimulation which seems to be present in lymphocytes from both old and Down's syndrome subjects.     

Dissociation and unfolding of bovine odorant binding protein at acidic pH

Dissociation of bovine odorant binding protein (bOBP) dimers to monomers at pH 2.5 has been confirmed through size exclusion chromatography experiments. Moreover, structural and binding properties of the acidic monomer and neutral dimer have been compared using a combination of experimental (circular dichroism and fluorescence) and computational (molecular dynamics) techniques. The secondary and tertiary structures of bOBP are largely maintained at acidic pH, but molecular dynamics simulations suggest the loop regions (N-terminal residues, Omega-loop and C-terminal segments) are more relaxed and Phe36 and Tyr83 residues are involved in the regulation of the binding cavity entrance. The formation of a molten globule state at acidic pH, suggested by the strong enhancement of fluorescence of 8-anilino-1-naphtalenesulphonic acid (ANS), is not confirmed by any significant change in the near UV circular dichroism spectrum. Functionality measurements, deduced from the interaction of bOBP with 1-amino-anthracene (AMA), show that the binding capacity of the protein at acidic pH is preserved, though slightly looser than at neutral pH. Unfolding of acidic bOBP, induced by guanidinium chloride (GdnHCl), was investigated by means of CD spectroscopy, steady state fluorescence, fluorescence anisotropy and light scattering. The stability of the acidic monomer is lower than that of the neutral dimer, owing to the loss of the swapping interactions, but renaturation is completely reversible. Finally, in contrast with the neutral dimer, at low denaturant concentration some aggregation of the acidic monomer, which vanishes before the unfolding transition, has been observed.     

Rosmarinic acid content in basil plants grown <Emphasis Type="Italic">in vitro</Emphasis> and in hydroponics

The accumulation of selected caffeic acid derivatives (CADs), in particular rosmarinic acid (RA), was investigated in different tissues (leaves, roots and plantlet shoots) of sweet basil (Ocimum basilicum L.) plants grown either in vitro or in hydroponic culture (floating system) under greenhouse conditions. Two cultivars with green leaves (Genovese and Superbo) and one with purple leaves (Dark Opal) were tested. The content of CADs in HCl-methanol extracts was determined by HPLC. LC-MS and LC-MS-MS were used to confirm the identification of the metabolites of interest. Apart from rosmarinic acid (RA) and a methylated form of this substance, no other CADs were detected at significant level in any of the analyzed samples. The content of RA ranged approximately from 4 to 63 mg/g DW, depending on the growing system. The highest RA content was found during the in vitro multiplication, in the acclimatized plants and in the roots of hydroponically-grown seedlings at full bloom. In vitro, 6-benzyladenine reduced the accumulation of RA in purple-leaf Dark Opal cultivar, but an opposite effect of this growth regulator was observed in the green-leaf genotypes. Our findings suggest the possibility to scale-up RA production by means of in vitro or hydroponic culture of sweet basil.     

Isolated Short Fetal Femur Length in the Second Trimester and the Association with Adverse Perinatal Outcome: Experiences from a Tertiary Referral Center

Objective

To determine the association between isolated mid-trimester short fetal femur length and adverse perinatal outcome.

Methods

This is a retrospective cohort study of patients with singleton gestations routinely assessed by second trimester ultrasound examination during 2006-2013. A fetal isolated short femur was defined as a femur length (FL) below the 5^th percentile in a fetus with an abdominal circumference greater than the 10^th percentile. Cases of aneuploidy, skeletal dysplasia and major anomalies were excluded. Primary outcomes of interest included the risk of small for gestational age neonates, low birth weight and preterm birth (PTB). Secondary outcome parameters were a 5-min Apgar score less than 7 and a neonatal intensive care unit admission. A control group of 200 fetuses with FL ≥ 5th percentile was used to compare primary and secondary outcome parameters within both groups. Chi-square and Student’s t-tests were used where appropriate.

Results

Out of 608 eligible patients with a short FL, 117 met the inclusion criteria. Isolated short FL was associated with an increased risk for small for gestational age (19.7% versus 8.0%, p = 0.002) neonates, low birth weight (23.9% versus 8.5%, p<0.001), PTB (19.7% versus 6.0%, p<0.001) and neonatal intensive care unit admissions (13.7% versus 3.5%, p = 0.001). The incidence of a 5-min Apgar score less than 7 was similar in both groups.

Conclusion

Isolated short FL is associated with a subsequent delivery of small for gestational age and Low birth weight neonates as well as an increased risk for PTB. This information should be considered when counseling patients after mid-trimester isolated short FL is diagnosed.