Thrombin-induced secretion of serotonin from platelets can occur in seconds

The platelet release reaction was studied by a new quenched-flow approach. Platelets labeled with 14C-serotonin were reacted for short times (up to 5 sec) with thrombin and then quenched with glutaraldehyde or paraformaldehyde. Serotonin secretion began within 1 sec and was nearly complete by 4 sec. Aggregation recorded by a resistive-particle counter was similarly fast. Therefore, the quenched-flow system reveals that serotonin secretion can occur more rapidly than estimated in earlier studies.     

Calcification in undifferentiated abdominal malignancies.

Four cases of undifferentiated abdominal malignancy are reported with a pattern of bizarre dystrophic calcifications. All of the cases had confusing histology and were long-term survivors; two being still alive. We feel that the radial, conglomerate masses of calcification, which do not conform to any organ, should suggest the diagnosis of an undifferentiated malignancy. The differential diagnosis and distinction from other causes of abdominal calcification is discussed.     

Identification of sites responsible for the potentiating effect of niflumic acid on ClC-Ka kidney chloride channels

Background and purpose:

ClC-K kidney Cl⁻ channels are important for renal and inner ear transepithelial Cl⁻ transport, and are potentially interesting pharmacological targets. They are modulated by niflumic acid (NFA), a non-steroidal anti-inflammatory drug, in a biphasic way: NFA activates ClC-Ka at low concentrations, but blocks the channel above ∼1 mM. We attempted to identify the amino acids involved in the activation of ClC-Ka by NFA.

Experimental approach:

We used site-directed mutagenesis and two-electrode voltage clamp analysis of wild-type and mutant channels expressed in Xenopus oocytes. Guided by the crystal structure of a bacterial CLC homolog, we screened 97 ClC-Ka mutations for alterations of NFA effects.

Key results:

Mutations of five residues significantly reduced the potentiating effect of NFA. Two of these (G167A and F213A) drastically altered general gating properties and are unlikely to be involved in NFA binding. The three remaining mutants (L155A, G345S and A349E) severely impaired or abolished NFA potentiation.

Conclusions and implications:

The three key residues identified (L155, G345, A349) are localized in two different protein regions that, based on the crystal structure of bacterial CLC homologs, are expected to be exposed to the extracellular side of the channel, relatively close to each other, and are thus good candidates for being part of the potentiating NFA binding site. Alternatively, the protein region identified mediates conformational changes following NFA binding. Our results are an important step towards the development of ClC-Ka activators for treating Bartter syndrome types III and IV with residual channel activity.     

4α-phorbol 12,13-didecanoate activates cultured mouse dorsal root ganglia neurons independently of TRPV4

Background and Purpose

The Ca²⁺-permeable cation channel TRPV4 is activated by mechanical disturbance of the cell membrane and is implicated in mechanical hyperalgesia. Nerve growth factor (NGF) is increased during inflammation and causes mechanical hyperalgesia. 4α-phorbol 12,13-didecanoate (4αPDD) has been described as a selective TRPV4 agonist. We investigated NGF-induced hyperalgesia in TRPV4 wild-type (+/+) and knockout (–/–) mice, and the increases in [Ca²⁺]_i produced by 4αPDD in cultured mouse dorsal root ganglia neurons following exposure to NGF.

Experimental Approach

Withdrawal thresholds to heat, von Frey hairs and pressure were measured in mice before and after systemic administration of NGF. Changes in intracellular Ca²⁺ concentration were measured by ratiometric imaging with Fura-2 in cultured DRG and trigeminal ganglia (TG) neurons during perfusion of TRPV4 agonists.

Key Results

Administration of NGF caused a significant sensitization to heat and von Frey stimuli in TRPV4 +/+ and –/– mice, but only TRPV4 +/+ mice showed sensitization to noxious pressure. 4αPDD stimulated a dose-dependent increase in [Ca²⁺]_i in neurons from +/+ and –/– mice, with the proportion of responding neurons and magnitude of increase unaffected by the genotype. In contrast, the selective TRPV4 agonist GSK1016790A failed to stimulate an increase in intracellular Ca²⁺ in cultured neurons. Responses to 4αPDD were unaffected by pretreatment with NGF.

Conclusions and Implications

TRPV4 contributes to mechanosensation in vivo, but there is little evidence for functional TRPV4 in cultured DRG and TG neurons. We conclude that 4αPDD activates these neurons independently of TRPV4, so it is not appropriate to refer to 4αPDD as a selective TRPV4 agonist.     

The Use of Condensational Growth Methods for Efficient Drug Delivery to the Lungs during Noninvasive Ventilation High Flow Therapy

Purpose

The objective of this study was to evaluate the delivery of nasally administered aerosols to the lungs during noninvasive ventilation using controlled condensational growth techniques.

Methods

An optimized mixer, combined with a mesh nebulizer, was used to generate submicrometer aerosol particles using drug alone (albuterol sulfate) and with mannitol or sodium chloride added as hygroscopic excipients. The deposition and growth of these particles were evaluated in an adult nose-mouth-throat (NMT) model using in vitro experimental methods and computational fluid dynamics simulations.

Results

Significant improvement in the lung dose (3–4× increase) was observed using excipient enhanced growth (EEG) and enhanced condensational growth (ECG) delivery modes compared to control studies performed with a conventional size aerosol (~5 μm). This was due to reduced device retention and minimal deposition in the NMT airways. Increased condensational growth of the initially submicrometer particles was observed using the ECG mode and in the presence of hygroscopic excipients. CFD predictions for regional drug deposition and aerosol size increase were in good agreement with the observed experimental results.

Conclusions

These controlled condensational growth techniques for the delivery of submicrometer aerosols were found to be highly efficient methods for delivering nasally-administered drugs to the lungs.     

Comparing accommodative function between the dominant and non-dominant eye

Background

To compare the accommodative amplitude (AA), facility (AF), and lag between dominant and non-dominant eyes.

Methods

Seventy students [mean (SD) age: 21.2 (1.7) years, range 18–25] from Zahedan University of Medical Sciences were selected. Retinoscopy and subjective refraction was used to determine the refractive error. The hole-in-the card method was used to determine eye dominance. The accommodative amplitude (AA) was measured in the dominant and non-dominant eye using the push-up method, and accommodative facility (AF) using ±2.00 dioptre flipper lenses at 40 cm. Accommodative lag was determined using monocular estimate method (MEM) retinoscopy at 40 cm.

Results

The right eye was dominant in 53 subjects (75.7 %). There was no significant difference in refractive error (sphere, cylinder, and spherical equivalent) between dominant and non-dominant eyes. The mean (SD) for the AA, AF, and lag in dominant eyes was 12.48 (2.56) dioptres, 12.45 (4.83) cycles per minute, and 0.80 (0.27) dioptres respectively. The mean (SD) for the AA, AF, and lag in non-dominant eyes was 12.16 (2.37) dioptres, 12.20 (4.88) cycles per minute, and 0.83 (0.28) dioptres respectively. The mean (SD) difference in AA, AF, and lag between dominant and non dominant eyes was 0.32 (0.75) dioptres (P?=?0.001), 0.25 (1.05) cycles per minute (P?=?0.04), and ?0.02 (0.11) dioptres (P?=?0.10) respectively. The AA and AF was statistically better (P?<?0.05) in the dominant eye group than in the non-dominant eye group. These data provided little evidence of any difference in the accommodative lag between dominant and non-dominant eyes (P?>?0.05).

Conclusion

The right eye was dominant in 76 % of subjects. Superior AA and AF was found in the dominant eye as determined by hole-in-the card method in young healthy adults, although these differences are perhaps not of clinical significance (<0.50 dioptres and <2 cycles per minute).     

Characterization of the IVS-II-821 (A>C) (HBB: c.316-30A>C) Mutation in a β-Thalassemia Phenotype in Iran

β-Thalassemia (β-thal) is the most frequently observed hereditary blood disorder that results from genetic defects causing deficient synthesis of hemoglobin (Hb) polypeptide chains. Detecting thalassemia mutations are necessary for prenatal diagnosis (PND) programs leading a better quality of life for the patients, as well as a reduction in the cost of their medical care. There are more than 900 different genomic mutations of the β-globin gene described in the human hemoglobin variant (HbVar) database. In this study, we identified a mid-intronic mutation at IVS-II-821 (A>C) (HBB: c.316-30A>C) position in the HBB gene of an Iranian proband and two of her siblings that was associated with β-thal clinical features. Direct DNA sequence analysis was performed by mutation scanning of the β-globin gene. Based on the observed β-thal phenotype and bioinformatics analysis results, we concluded that this β-globin gene mutation was associated with a mild phenotype of β-thal through activating potential splice sites by creating exonic splicing enhancers (ESEs), exon-identity element (EIE) and exonic splicing regulatory sequences (ESRs) sites.     

THE IMPACT OF PRE-CLERKING CLINICS ON SURGICAL OPERATION CANCELLATIONS: A PROSPECTIVE AUDIT

Pre-clerking of all patients undergoing elective general surgical operations was introduced at our hospital in an attempt to reduce an unacceptably high operation cancellation rate. A prospective audit has been performed on the effect of this policy on the cancellation rate. Before the introduction of pre-clerking there was a marked seasonal variation in the number of patients who failed to attend for surgery, which could be explained by absence on holiday. This seasonal variation disappeared after the start of pre-clerking clinics, but there has been no reduction in the number of cancellations for medical reasons.