A pharmacological examination of venom from the Papuan taipan (Oxyuranus scutellatus canni).

The Papuan taipan (Oxyuranus scutellatus canni) is the third most venomous terrestrial snake in the world, however, little is know about the pharmacology of the venom. In the chick biventer cervicis muscle, venom (10 μg/ml) abolished nerve-mediated twitches (time to 90% inhibition (t₉₀) 44±5 min, n=9). This inhibition was unaffected by prior incubation of the venom with the phospholipase A inhibitor 4-bromophenacyl bromide (4-BPB; 0.72 mM) (t₉₀ 48±7 min, n=8). The mouse phrenic nerve diaphragm preparation displayed greater sensitivity to venom (10 μg/ml) (t₉₀ 25±1 min, n=6). In the chick biventer muscle, venom (10 μg/ml) significantly inhibited responses to acetylcholine (1 mM) and carbachol (20 μM), but not KCl (40 mM), indicating activity at post-synaptic nicotinic receptors. Venom (10 μg/ml) did not affect direct muscle stimulation. Venom (3–30 μg/ml) produced dose-dependant contractions of the guinea-pig ileum. Contractile responses were significantly inhibited by indomethacin (1 μM) or prior incubation of the venom with 4-BPB (0.72 mM) indicating involvement of a PLA component. In rat phenylephrine (0.3 μM) precontracted aortae, venom (3–100 μg/ml) produced endothelium-independent relaxation which was unaffected by prior incubation of venom (30 μg/ml) with 4-BPB (0.72 mM). In anaesthetised rats, 10 μg/kg (i.v.) venom produced rapid respiratory and cardiovascular collapse while 5 μg/kg (i.v.) venom produced only a small transient decrease in mean arterial blood pressure. Prior administration of 5 μg/kg (i.v.) venom enabled subsequent administration of 10 and 100 μg/kg (i.v.) venom without respiratory or cardiovascular collapse. Further work is required to identify specific toxins with the above pharmacological activity.     

The effects of antivenom on the in vitro neurotoxicity of venoms from the taipans Oxyuranus scutellatus, Oxyuranus microlepidotus and Oxyuranus scutellatus canni.

The venoms of the inland (Oxyuranus microlepidotus), coastal (O. scutellatus) and Papuan (O. s. canni) taipans are among the most potent in the world. The present study compared the in vitro neurotoxic effects of these venoms and the protective effects of taipan antivenom. Venom (10 μg/ml) from all three snakes abolished nerve-mediated twitches of the chick biventer cervicis muscle preparation with the following rank order of potency (based on the time taken to inhibit 90% of the twitch response; t₉₀): O. microlepidotus (27±3 min)>O. scutellatus (42±3 min)=O. S. canni (48±5 min). This inhibitory effect of all three venoms was primarily postsynaptic in origin as evidenced by the inhibition of responses to exogenous acetylcholine (ACh; 1 mM) and carbachol (CCh; 20 μM), but not potassium chloride (40 mM). In contrast, the presynaptic neurotoxins taipoxin (3 μg/ml) and paradoxin (3 μg/ml) abolished nerve-mediated twitches without producing a significant effect on contractile responses to exogenous agonists. Prior incubation of the tissue with taipan antivenom (1 unit/ml for 10 min) markedly attenuated the inhibitory effects of taipoxin (3 μg/ml) and paradoxin (3 μg/ml), as well as O. scutellatus (10 μg/ml) and O. s. canni (10 μg/ml) venom. However, in the presence of antivenom, O. microlepidotus venom (10 μg/ml) still abolished nerve-mediated twitches and responses to ACh and CCh. The results of the current study indicate that taipan antivenom, raised against O. scutellatus venom, is effective, in vitro, against the neurotoxic effects of venom from the Papuan and coastal taipans, as well as the presynaptic effects of venom from the inland taipan. However, the antivenom appears less effective against the postsynaptic effects of the latter. It is possible that inland taipan venom contains a component not neutralised by the antivenom which may contribute to the extreme potency of this venom.     

Typing classical polymorphisms by real‐time PCR: Analysis of the GPT and ALAD protein polymorphisms in the Jewish populations

Expanding the already extensive data array on classical polymorphisms in new populations is beneficial to full characterization of the intricate relationships between human populations. This can be done in a short time on a large‐scale by using real‐time PCR. The red‐cell enzymes Δ‐aminolevulinate dehydratase (ALAD) and glutamate pyruvate transaminase (GPT) have each two alleles that can be separated by protein electrophoresis. They were previously characterized by protein electrophoresis in some of the Jewish populations. We designed primers and probes for fluorescence resonance energy transfer detection of the ALAD and GPT alleles to type Jewish populations including some in which these polymorphisms have not been studied in the past. The methods described here allow fast and efficient determination of these common classical polymorphisms. When used in large‐scale population studies, these methods are not only faster, but also cheaper than amplification followed by restriction enzyme digestion and gel electrophoresis. The frequency of ALAD*1 among Jews ranges from 0.781 to 0.960 and that of GPT*1 from 0.412 to 0.773. A maximum likelihood tree based on the ALAD and GPT polymorphisms and five other red‐cells enzymes polymorphisms depicts quite accurately the known historic, anthropological, and geographic relations between the Jewish populations. Am. J. Hum. Biol., 2008. © 2008 Wiley‐Liss, Inc.     

Nucleotide variants within the IQGAP1 gene in diffuse-type gastric cancers

IQGAP1 is recognized as a negative regulator of cell-cell adhesion at adherens junctions in several cell types, including gastric mucosal cells. The histopathologic appearance of diffuse gastric carcinoma is defined by non- or poorly cohesive tumor cells, indicating abnormal intercellular adhesion. Hence, we screened 38 gastric cancers for activating point mutations in IQGAP1. In 2 of the 33 diffuse gastric cancers, there was a missense nucleotide change predicted to alter the amino acid sequence in the GAP-related domain, which includes part of the binding site for the activated small G proteins Cdc42 and Rac1. Many intronic IQGAP1 gene changes were observed, and several occurred more frequently in diffuse-type gastric cancers than in intestinal-type gastric cancers. A highly variable pentanucleotide repeat was identified in the final intron of IQGAP1. The most expanded six-repeat sequence was present exclusively in diffuse-type gastric cancers. Additionally, 19 diffuse cases and two intestinal cases exhibited silent coding region nucleotide alterations. Taken together, our results suggest that IQGAP1 coding sequence mutations are not a frequent event in gastric cancer, but do occur in a subset of diffuse-type gastric carcinomas. Additional studies analyzing other proteins involved in cell adhesion may lead to a better molecular understanding of the histopathologic appearance of diffuse gastric cancers.     

Oximeter's acquisition parameter influences the profile of respiratory disturbances

STUDY OBJECTIVES: Pulse oximetry (Sp02) is a key parameter monitored during polysomnographic studies, and different acquisition settings can be employed to obtain this data. The purpose of this study was to determine if the use of different settings would significantly influence scoring of respiratory disturbance events (RDE). DESIGN: Prospective study SETTING: Sleep Disorders Center - community PATIENTS: 30 patients had three identical oximeters simultaneously attached to the digits during polysomnography, each placed in a different recording setting: 3, 6 and 12 seconds. INTERVENTIONS: None. MEASUREMENTS: RDEs were identified by changes in snoring and flow then sub-categorized as RDE0, RDE1-2 and/or RDE3 if less than 1%, greater than 1 but less than 3%, and 3% or greater oxyhemoglobin desaturation occurred. Each event was given three labels according to the level of desaturation seen on each oximetry tracing. RESULTS: Significant differences in the mean frequency of RDE types at each recording setting were noted (p < .001). A survey of sleep practitioners revealed changes in clinical behavior when presented examples of such differences. CONCLUSION: These data confirm the impact of different oximetric recording settings on the profile of RDEs and the importance of reporting such acquisition settings in studies of sleep disordered breathing.     

Coexistence of peptide immunoreactivity in sensory neurons of the cat

The coexistence of the neuropeptides substance P, cholecystokinin, somatostatin and vasoactive intestinal polypeptide in cat sensory neurons has been examined using peroxidase-anti-peroxidase immunocytochemistry. Attempts were also made to locate cells containing bombesin, neurotensin, [Met]enkephalin and [Leu]enkephalin but no immunoreactivity was found when antisera to these peptides was used. Cells in the dorsal root ganglia were studied by cutting 5 microns serial wax sections or 15 microns cryostat sections. Coexistence was established by applying the antiserum to each peptide to serially adjacent 5 microns sections and establishing the presence of peptide-like immunoreactivity in each of 4 different sections through a single cell. Results showed that the distribution and combinations of coexistence of these neuropeptides in the cat is extremely complex; three and sometimes all four antisera showing immunoreactivity with a single cell. About 21% of all ganglion cells contained some immunoreactivity but there were certainly some small cells which did not contain any immunoreactivity. The coexistence of these peptides differed markedly from that previously reported in the rat suggesting that interspecific differences in the neuropeptide content of cells might be much greater than they are for classical neurotransmitters. The results are discussed in relation to the possible role of neuropeptides and the regulation of their production by sensory neurons.     

The satiating efficiency of foods

Experiments were undertaken to test the general hypothesis that some foods are more satiating than others, to find a mechanism for their differential satiating efficiencies, and to determine whether certain soups had a high enough satiating efficiency to recommend their addition to a meal as a way of reducing total caloric intake of that meal. In the first experiment it was found that intake of a test meal was lower after a large preload of tomato soup than after a small preload in women, but not in men. However, the total energy intake (soup plus test meal) was no less with meals which included the large soup preload than it was with meals that did not include a preload. Therefore adding a normal portion of tomato soup to a meal would not reduce its total energy intake. We noted the interesting incidential finding that total energy intake (i.e., preload plus test meal) of the meals which contained the larger amount of soup was less than the total energy intake of the meals which contained a combination of crackers, jelly, and juice. In the second experiment we confirmed this finding by showing that when equal weights of tomato soup preloads and a preload of crackers, cheese, and apple juice, which contains more energy, were given, total energy intake was less in meals which included soup. Therefore, substituting tomato soup for a more calorically dense first course could reduce total energy intake of that meal. In the third experiment, the hypothesis suggested by the second was confirmed. Two soups were more satiating than crackers, cheese, and juice. When two calorie levels were used for each preload, it was shown that calorie for calorie, these soups decreased intake of the test meal more than crackers, cheese, and juice. In the fourth experiment we showed that the mechanism for this differential satiating efficiency is not readily attributable to either bulk related factors or fat content. We suggest that the differential satiating efficiencies are related to differences in nutrient dispersion, orosensory cues, or temperature. Finally, reductions in intake were accompanied by reductions in the initial rate of eating and not by increases in the rate of deceleration. This reduction was small but consistent and suggests that foods which are more satiating reduce intake by decreasing desire to eat (i.e., hunger), not by accelerating the onset of meal termination (i.e., satiety). In fact the duration of meals was unaffected by the preloads.     

Familial mediterranean fever: The segregation of four different mutations in 13 individuals from one inbred family: Genotype–phenotype correlation and intrafamilial variability

Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurring attacks of fever and serositis. Six sequence alterations (M694V, V726A, K695R, M680I, M694I, and E148Q), in the MEFV gene, account for the majority of FMF chromosomes. Differences in the clinical expression have been mainly attributed to MEFV allelic heterogeneity. Homozygotes for the M694V mutation have a more severe form of the disease and more frequently demonstrate articular and renal complications. The clinical manifestations associated with mutation M680I are considered less severe. Mutations E148Q, K695R and V726A have reduced penetrance, and many individual homozygotes or compound heterozygotes for these mutations remain asymptomatic. Here we report on one inbred family with 13 individuals (one grandparent, three parents, and nine grandchildren), either homozygotes or compound heterozygotes, for one or two of four mutations (V726A, M694V, M680I, and K695R). Three parents and one grandparent who each carried two mutated alleles remained asymptomatic. Of nine grandchildren who were compound heterozygotes for two mutations in the MEFV gene, only those with either the M694V/V726A or the M694V/M680I genotypes manifested the disease, bearing further evidence to the severity of mutation M694V in individuals sharing a similar genetic and environmental background. Nevertheless, one father and one grandmother who carried the M694V/V726A compound heterozygous genotype were symptom‐free, while the four grandchildren with the same genotype manifested the disease from early age, providing further evidence for the role of additional environmental and genetic modifiers. The occurrence of four different mutations in two sets of consanguineous parents merits consideration per se. © 2002 Wiley‐Liss, Inc.