Phylogeography, intraspecific structure and sex-biased dispersal of Dall's porpoise, Phocoenoides dalli, revealed by mitochondrial and microsatellite DNA analyses

Mitochondrial DNA (mtDNA) control-region sequences and microsatellite loci length polymorphisms were used to estimate phylogeographical patterns (historical patterns underlying contemporary distribution), intraspecific population structure and gender-biased dispersal of Phocoenoides dalli dalli across its entire range. One-hundred and thirteen animals from several geographical strata were sequenced over 379 bp of mtDNA, resulting in 58 mtDNA haplotypes. Analysis using F(ST) values (based on haplotype frequencies) and phi(ST) values (based on frequencies and genetic distances between haplotypes) yielded statistically significant separation (bootstrap values P < 0.05) among most of the stocks currently used for management purposes. A minimum spanning network of haplotypes showed two very distinctive clusters, differentially occupied by western and eastern populations, with some common widespread haplotypes. This suggests some degree of phyletic radiation from west to east, superimposed on gene flow. Highly male-biased migration was detected for several population comparisons. Nuclear microsatellite DNA markers (119 individuals and six loci) provided additional support for population subdivision and gender-biased dispersal detected in the mtDNA sequences. Analysis using F(ST) values (based on allelic frequencies) yielded statistically significant separation between some, but not all, populations distinguished by mtDNA analysis. R(ST) values (based on frequencies of and genetic distance between alleles) showed no statistically significant subdivision. Again, highly male-biased dispersal was detected for all population comparisons, suggesting, together with morphological and reproductive data, the existence of sexual selection. Our molecular results argue for nine distinct dalli-type populations that should be treated as separate units for management purposes.     

Entamoeba histolytica: mechanism of decrease of virulence of axenic cultures maintained for prolonged periods

Intraportal injection of non-virulent E. histolytica (derived from prolonged axenic culture of virulent E. histolytica) strain HM1-IMSS in normal hamsters results in no liver lesions and disappearance of the parasites 48-72 h after injection. Viability of non-virulent E. histolytica after 2 h of in vitro incubation in either fresh or decomplemented hamster serum is the same as control virulent E. histolytica (50-90%). In hamsters made leukopenic, or both leukopenic+hypocomplementemic, or hypocomplementemic+sephadex microspheres (to produce focal liver ischemia) intraportally injected non-virulent E. histolytica cause no lesions and disappear after 24 h. In addition, neither hypocomplementemia nor immunosuppression with cyclosporin A prolonged the survival of non-virulent E. histolytica. Methyl prednisolone treatment of hamsters resulted in survival of large numbers of non-virulent E. histolytica in the liver, with little inflammation and minimal tissue damage, for up to 7 days. Inflammatory cells (macrophages) would appear to be chiefly responsible for elimination of non-virulent E. histolytica. Parallel experiments with E. dispar suggest a different mechanism for its non-pathogenicity.     

UNOBSERVED KILL OF NURSING DOLPHIN CALVES IN A TUNA PURSE-SEINE FISHERY

The kill of dolphins ( Stenella attenuata and S. longirostris ) in the eastern tropical Pacific tuna purse-seine fishery has been underestimated because of unobserved deaths of nursing calves due to separation from their mothers during fishing. Based on an analysis of dolphins killed from 1973 to 1990, and depending on the length at which calves are assumed to become independent, there was a deficit of calves relative to the number of lactating females killed in 24%–32% of 1,847 spotted-dolphin sets and in 13%–19% of 563 spinner-dolphin sets. We found a deficit of 0.31–0.45 spotted dolphin and 0.15–0.26 spinner dolphin calves per set. If these missing calves were added to the observed kill, it would represent an increase in the kill of 10%–15% for spotted dolphins and 6%–10% for spinner dolphins in the sets we examined. We did not attempt to estimate the actual number of unobserved calf deaths due to purse-seine fishing on dolphins, either in the sets we examined or in all dolphin sets. The actual number of unobserved calf deaths is likely to be higher than the calf deficit we found. Separation of dolphin mothers from calves could occur at any of several points in the fishing process, but most of these would be invisible to us and not produce a calf deficit. Estimation of the actual number of unobserved calf deaths would require further information on how frequently permanent mother-calf separations occur, the fraction of calves that survive after separation, the fraction of lactating females set upon that are carrying calves, and the fraction of calves killed that are actually related to lactating females killed in the same set. In any case, the observation of a calf deficit indicates that the reported dolphin kill fails to measure the full impact of purse-seine fishing on spotted and spinner dolphin populations.     

Preparation of chiral derivatives of beta-Ala containing the alpha-phenylethyl group: useful starting materials for the asymmetric synthesis of beta-amino acids

The use of microwave (MW) irradiation for the condensation reaction between acetophenone and alpha-phenylethylamine to prepare (R,R)-bis[alpha-phenylethyl]amine results in significantly reduced reaction times relative to the use of conventional heating. In this protocol, a secondary amine, (R,R)-bis(alpha-phenylethyl)amine is treated with acryloyl chloride to afford conjugated amide N,N-bis[(R)-alpha-phenylethyl]prop-2-enamide, (R,R)-3. 1,4-Addition of alpha-phenylethylamine to unsaturated (R,R)-3 affords propanamide N,N-Bis[(R)-alpha-phenylethyl]-3-N-[(S)-alpha-phenylethyl]amino-propanamide, (R,R,S)-4, which can be alkylated with high diastereoselectivity to give derivative N,N-Bis[(R)-alpha-phenylethyl]-3-N'-[(S)-alpha-phenylethyl]amino-propanamide, (R,R,S,S)-5. Hydrogenolysis of (R,R,S,S)-5 catalyzed by palladium hydroxide and final hydrolysis (4 N HCl) resulting in the formation of (S)-alpha-benzyl-beta-alanine, (S)-7, is facilitated by MW irradiation. The use of MW irradiation in this step prevents racemization of the desired amino acid. The present protocol constitutes one of the simplest strategies for the asymmetric synthesis of biologically relevant alpha-substituted-beta-amino acids since it takes advantage of inexpensive, commercially available beta-Ala and either (R)- or (S)-alpha-phenylethylamine as chiral auxiliary. The required time for this protocol is approximately 90 h, which can be carried out in 5 d.     

Entamoeba histolytica: apoptosis induced in vitro by nitric oxide species

Apoptosis has been described in some parasites like Leishmania, Trypanosoma, and Trichomonas. This phenomenon has not been observed yet in Entamoeba histolytica. This work analyzed the in vitro effect of sodium nitroprusside, sodium nitrite and sodium nitrate (NOs) on E. histolytica apoptosis. Parasites incubated for 1h with NOs revealed apoptosis 6h later (95% viability), demonstrated by YOPRO-1, TUNEL, DNA fragmentation and low ATP levels. The caspase inhibitor Z-VAD-FMK inhibited total intracellular cysteine protease activity (CPA) but had no effect on apoptosis. When treated with NOs some amebic functions like complement resistance and hemolytic activity decreased but CPA and erythrophagocytosis remained unchanged. After treatment in vitro with NOs, parasite death was almost complete at 24h; but when injected into hamster livers they disappeared in less than 6h. These results show that apoptosis is induced in vitro by NOs in E. histolytica and renders them incapable of surviving in hamster's livers.     

Does Cataract Surgery Alleviate Poverty? Evidence from a Multi-Centre Intervention Study Conducted in Kenya,the Philippines and Bangladesh

Background

Poverty and blindness are believed to be intimately linked, but empirical data supporting this purported relationship are sparse. The objective of this study is to assess whether there is a reduction in poverty after cataract surgery among visually impaired cases.

Methodology/Principal Findings

A multi-centre intervention study was conducted in three countries (Kenya, Philippines, Bangladesh). Poverty data (household per capita expenditure – PCE, asset ownership and self-rated wealth) were collected from cases aged ≥50 years who were visually impaired due to cataract (visual acuity<6/24 in the better eye) and age-sex matched controls with normal vision. Cases were offered free/subsidised cataract surgery. Approximately one year later participants were re-interviewed about poverty. 466 cases and 436 controls were examined at both baseline and follow-up (Follow up rate: 78% for cases, 81% for controls), of which 263 cases had undergone cataract surgery (“operated cases”). At baseline, operated cases were poorer compared to controls in terms of PCE (Kenya: $22 versus £35 p = 0.02, Bangladesh: $16 vs $24 p = 0.004, Philippines: $24 vs 32 p = 0.0007), assets and self-rated wealth. By follow-up PCE had increased significantly among operated cases in each of the three settings to the level of controls (Kenya: $30 versus £36 p = 0.49, Bangladesh: $23 vs $23 p = 0.20, Philippines: $45 vs $36 p = 0.68). There were smaller increases in self-rated wealth and no changes in assets. Changes in PCE were apparent in different socio-demographic and ocular groups. The largest PCE increases were apparent among the cases that were poorest at baseline.

Conclusions/Significance

This study showed that cataract surgery can contribute to poverty alleviation, particularly among the most vulnerable members of society. This study highlights the need for increased provision of cataract surgery to poor people and shows that a focus on blindness may help to alleviate poverty and achieve the Millennium Development Goals.     

Titles of related papers in other sections

The effect of hypoglycemic drugs chlorpropamide and phenformin has been tested on isolated liver plasma membranes as to their microviscosity parameters. Results reported suggest that both drugs are able to increase in vitro plasma membrane microviscosity in a dose-dependent way.