Augmentation in the infectivity of SV40 virus DNA by amphotericin B methyl ester

Summary The methyl ester of amphotericin B (100 µg/ml) enhanced the infectivity of SV40 virus DNA for CV-1 cells up to 300-fold. Amphotericin B methyl ester may prove useful for augmenting cellular penetration of macromolecules and for probing biological mechanisms.With 2 Figures     

A microperfusion investigation of bicarbonate secretion by the rat submaxillary gland

Summary Bicarbonate transport in the rat submaxillary main duct has been studied by microperfusion. Bicarbonate was concentrated in the duct lumen against an electrochemical gradient and the equilibrium concentration was estimated to be 56.5 mEq/l±3.1 (S.E.M.,n=11). The secretory mechanism could not be inhibited by 6 mMolar cyanide although such concentrations caused marked inhibition of both net sodium efflux and net potassium influx. Bicarbonate secretion in the main duct was not inhibited by acetazolamide whether applied from the duct lumen or given intravenously. Similarly, the drug was without effect on bicarbonate excretion by the intact gland even when maximum excretory rates had been induced with carbachol. It was concluded that catalytic hydration of carbon dioxide to carbonic acid was not a rate-limiting step in the bicarbonate secretory process. The data did not permit a distinction to be made between a bicarbonate secretory processper se and a process of either H⁺ reabsorption or OH^– secretion.The parasympathomimetic agent, carbachol, when given parenterally was found to increase sharply the net influx of bicarbonate into the microperfused main duct as well as to reduce net sodium efflux and net potassium influx. Previously it had been postulated that final saliva was formed in two stages. First a plasma-like primary secretion was formed at a rate depending on the degree of stimulation, and second, the primary secretion was modified in the gland duct system by reabsorptive and secretory processes whose transport rates were presumed to be independent of the degree of stimulus. It now becomes necessary to postulate that stimulation can act on electrolyte transport at both primary and secondary levels; at present, however, no data are available to show whether appreciable net water influx can ever occur at the secondary level.Preliminary reports of this study have been presented to the Australian Physiological and Pharmacological Society [28, 31].     

Relationship between X-inactivation and clinical involvement in Fabry heterozygotes. Eleven novel mutations in the α-galactosidase A gene in the Czech and Slovak population

We have identified 21 different -galactosidase A gene (GLA) mutations in 22 unrelated Czech and Slovak families with Fabry disease. Eleven of these mutations were novel (point mutations D93N, A135V, D155H, G171R, Q280K, G360S, Q330X, splicing errors c.194ins14, c.801ins36 and deletions c.674_732del59, g.3405_6021del2617). Genotyping of family members for family-specific mutations revealed 55 heterozygotes that manifested clinical symptoms of different severity. To examine the contribution of X-inactivation skewing to disease manifestation in Fabry heterozygotes, we have adopted the Mainz severity scoring scheme and compared the score values with the X-inactivation status in 39 carriers in an age-dependent manner. The age-score trendline of Fabry females who had a predominantly inactivated X-chromosome bearing a wild-type GLA allele (10 of 38 females) was markedly steeper than in the rest of the cohort. One female carrier with an inactivated mutated allele had a low score value when compared to the other heterozygotes of the same age. These data suggest that X-inactivation is indeed a major factor determining the severity of clinical involvement in Fabry heterozygotes. There was a statistically significant difference between the severity score values of heterozygotes with random and non-random X-chromosome inactivation at the 5% level of significance. Further studies will show if the degree of the wildtype allele inactivation will be useful as a predictive marker of severity of phenotype in Fabry heterozygotes. Although the correlation between X-inactivation skewing and presentation of the disease in Fabry heterozygotes has previously been suggested in the literature, this report is among the first attempts to examine this relationship systematically.     

Results of unrelated cord blood transplant in fanconi anemia patients: risk factor analysis for engraftment and survival.

We retrospectively analyzed results of unrelated cord blood transplantation (UCBT) in 93 Fanconi anemia (FA) patients. Median age at transplantation was 8.6 years (1-45). The units transplanted were HLA-A, -B, or -DRB1 identical in 12 cases, 1 HLA mismatch in 35 cases, and 2 or 3 HLA differences in 45 cases. The median number of nucleated cells (NC) and CD34+ cells infused of recipient weight was 4.9x10(7)/kg and 1.9x10(5)/kg, respectively. Participating centers selected the preparative regimen of their choice, in 57 patients (61%), it included Fludarabine. Graft-versus-host disease (GVHD) prophylaxis consisted mostly of cyclosporine with prednisone. Cumulative incidence (CI) of neutrophil recovery was 60+/-5% at day +60. In multivariate analysis, Fludarabine containing regimen and NC infused>or=4.9x10(7)/kg were associated with higher probability of recovery. CI of grade II-IV acute and of chronic GVHD (aGVHD, cGVHD) was 32%+/-5% and 16%+/-4%, respectively. Overall survival (OS) was 40%+/-5%. In multivariate analysis, factors associated with favorable outcome were use of Fludarabine in the conditioning regimen, number of NC infused>or=4.9x10(7)/kg, and negative cytomegalovirus (CMV) serology in the recipient. In conclusion, factors easily modifiable such as donor selection and a Fludarabine-containing regimen can considerably improve survival in FA patients given a UCBT. These data are the basis for designing prospective protocols.     

Estimating relative physical workload using heart rate monitoring: a validation by whole-body indirect calorimetry

Measuring physical workload in occupational medicine is fundamental for risk prevention. An indirect measurement of total and relative energy expenditure (EE) from heart rate (HR) is widely used but it has never been validated. The aim of this study was to validate this HR-estimated energy expenditure (HREEE) method against whole-body indirect calorimetry. Twenty-four-hour HR and EE values were recorded continuously in a calorimetric chambers for 52 adult males and females (19–65 years). An 8-h working period was retained, comprising several exercise sessions on a cycloergometer at intensities up to 65% of the peak rate of oxygen uptake. HREEE was calculated with reference to cardiac reserve. A corrected HREEE (CHREEE) was also calculated with a modification to the lowest value of cardiac reserve. Both values were further compared to established methods: the flex-HR method, and the use of a 3rd order polynomial relationship to estimate total and relative EE. No significant difference was found in total EE when measured in a calorimetric chamber or estimated from CHREEE for the working period. A perfect linear and identity relationship was found between CHREEE and energy reserve values for intensities ranging from 15% to 65%. Relative physical workload can be accurately assessed from HR recordings when expressed in CHREEE between 15% to 65%, and EE can be accurately estimated using the CHREEE method.     

Codon optimization of the human papillomavirus 11 (HPV 11) L1 gene leads to increased gene expression and formation of virus-like particles in mammalian epithelial cells

The 505 amino acid L1 protein of the human papillomavirus type 11 (HPV 11) is the major capsid polypeptide that has been shown to self-assemble into virus-like particles (VLPs) in vivo and in vitro. While L1 is essential for viral infection, expression studies in mammalian cells have been hampered by different codon preference between the virus and its host. To optimize L1 gene expression in mammalian cells, we converted wild-type HPV 11 L1 (11 L1wt) codons to those more common in human genes. The modified HPV 11 L1 gene (11 L1h) generated protein levels that were at least 100-fold higher than those of wild-type HPV 11 L1, while no obvious differences were seen in the level of mRNA. HPV 11 L1 protein was detected in mammalian epithelial and fibroblast cells, by immunoblotting and indirect immunofluorescence (IIF) techniques. Unlike the situation in situ, IIF revealed the presence of L1 mainly at perinuclear sites. Virus-like particles assembled intranuclearly only to a low extent, as indicated by transmission electron microscopy. DNA vaccination using the HPV 11 L1h gene yielded a drastic increase in L1-specific antibody production in mice as compared to immunization with the wild-type gene.     

Energy intake during reproduction in captive common marmosets (Callithrix jacchus)

Among haplorhine primates, the highly specialized Callitrichidae (marmosets and tamarins) are expected to have comparatively high reproductive costs, a feature that might be related to the evolution of a cooperative breeding system. Costs of reproduction in captivity were investigated on the basis of changes in energy intake and body weight during pregnancy and lactation in pair-living female and male common marmosets (Callithrix jacchus). The experimental design had little effect on carrying behavior, food intake, and body weight of adults, but a negative transitory effect of offspring body weight. Increased energetic requirements during pregnancy did not result in a higher energy intake in females. During lactation, females increased their energy intake up to 100% and gradually lost weight, suggesting even higher costs. Extensive carrying behavior by males, on the other hand, did not result in an increased energy intake in males, or in changes in male body weight. It is suggested that, at least in captivity, increased energetic demands during reproduction are reduced by behavior allocations towards energetically less expensive behaviors.