Quality of bull spermatozoa after preparation by single-layer centrifugation

The present study aimed to evaluate the effect of single-layer centrifugation (SLC) through a species-specific colloid (Androcoll-B; patent pending, J. M. Morrell) on bull sperm quality. Computer-assisted sperm analysis of motility and flow cytometric analysis of sperm viability (SYBR-14/propidium iodide staining), chromatin integrity (acridine orange staining), reactive oxygen species production [Hoechst 33258-hydroethidine-2′,7′-dichlorodihydrofluorescein diacetate (HO-HE-DCFDA) staining], mitochondrial membrane potential (staining with JC-1 probe), and protein tyrosine phosphorylation (specific antibody staining) were performed on unselected and SLC-selected sperm samples. Single-layer centrifugation of bull spermatozoa resulted in the selection of a sperm population that had high mitochondrial membrane potential, a higher content of phosphorylated protein, and more reactive oxygen species than control samples. Sperm chromatin damage was lower in the SLC samples although sperm viability and motility did not differ between SLC samples and controls. These observations suggest that SLC of bull semen in a soybean-containing extender improved some, but not all, parameters of sperm quality.     

Tobacco-related disease mortality among men who switched from cigarettes to spit tobacco

Background

Although several epidemiological studies have examined the mortality among users of spit tobacco, none have compared mortality of former cigarette smokers who substitute spit tobacco for cigarette smoking (“switchers”) and smokers who quit using tobacco entirely.

Methods

A cohort of 116 395 men were identified as switchers (n = 4443) or cigarette smokers who quit using tobacco entirely (n = 111 952) when enrolled in the ongoing US American Cancer Society Cancer Prevention Study II. From 1982 to 31 December 2002, 44 374 of these men died. The mortality hazard ratios (HR) of tobacco‐related diseases, including lung cancer, coronary heart disease, stroke and chronic obstructive pulmonary disease, were estimated using Cox proportional hazards regression modelling adjusted for age and other demographic variables, as well as variables associated with smoking history, including number of years smoked, number of cigarettes smoked and age at quitting.

Results

After 20 years of follow‐up, switchers had a higher rate of death from any cause (HR 1.08, 95% confidence interval (CI) 1.01 to 1.15), lung cancer (HR 1.46, 95% CI 1.24 to 1.73), coronary heart disease (HR 1.13, 95% CI 1.00 to 1.29) and stroke (HR 1.24, 95% CI 1.01 to 1.53) than those who quit using tobacco entirely.

Conclusion

The risks of dying from major tobacco‐related diseases were higher among former cigarette smokers who switched to spit tobacco after they stopped smoking than among those who quit using tobacco entirely.Several epidemiological studies have examined morbidity and mortality among users of spit tobacco (spit‐tobacco users),^1,2 but none have compared the mortality of former cigarette smokers who substitute spit tobacco for cigarette smoking (“switchers”) to those of smokers who quit using tobacco entirely. Comprehensive reviews by the US Surgeon General,³ and the International Agency for Research on Cancer⁴ and others⁵ have concluded that evidence is sufficient that the use of spit tobacco causes several types of cancer in humans. Although the evidence linking use of spit tobacco to increased risk of cardiovascular diseases is limited,⁶ these products cause acute increases in heart rate and blood pressure, as well as long‐term adverse effects on blood pressure and lipid profiles.⁷We compared mortality of switchers to those of former cigarette smokers who quit using tobacco entirely among men enrolled in a large prospective cohort.     

Effects of Modification Sequence on Structures and Properties of Hydroxypropylated and Crosslinked Waxy Maize Starch

The effects of modification sequence on chemical structures and physicochemical properties of hydroxypropylated (HP) and crosslinked (XL) waxy maize starch were investigated. The physicochemical properties, including pasting, gelling, and thermal properties, were studied. The chemical structures of dual‐modified starches and their beta‐limit dextrins were characterized with high‐performance liquid chromatography. The HP‐XL starch had higher Brabender viscosity than did the XL‐HP starch at both pH 7 and 3; however, both showed similar gelling properties. Significantly higher onset and peak gelatinization temperatures, gelatinization enthalpy, and lower retrogradation were observed for the HP‐XL starch. The HP‐XL starch also exhibited significantly higher beta‐amylolysis limit and higher content of low molecular weight saccharides in its isoamylase‐debranched starch, suggesting its structure was more accessible to enzymatic attack than the XL‐HP starch. Structural analyses revealed different distribution patterns of modifying groups between the two modified starches. The results indicate that the modification sequence altered the susceptibility to enzymes, changed the locations of substitution, and modified the physicochemical properties of the HP and XL waxy maize starches.     

Neonatal treatment of rats with diethylstilboestrol (DES) induces stromal-epithelial abnormalities of the vas deferens and cauda epididymis in adulthood following delayed basal cell development

This study investigated whether transient, neonatal (days 2-12) treatment of rats with the potent oestrogen, diethylstilboestrol (DES), altered the structure of the cauda epididymis/vas deferens in adulthood, and if the changes observed related to altered development of basal cells in early puberty. Neonatal treatment with 10 microg DES resulted in the following during adulthood: (a) coiling of the normally straight initial vas deferens, (b) gross epithelial abnormalities, (c) 4-fold widening of the periductal non-muscle layer, (d) infiltration of immune cells across the epithelium into the lumen, and (e) reduction/absence of sperm from the vas deferens lumen. Amongst affected animals>75% exhibited reduced epithelial immunoexpression of androgen receptor and aberrant oestrogen receptor-alpha immunoexpression and 63% exhibited multi-layering of basal cells coincident with increased epithelial cell proliferation. None of the aforementioned changes occurred in rats treated neonatally with 0.1 microg DES.As basal cells play a key role in the development of epithelia such as that in the epididymis and vas deferens, we went on to investigate if neonatal DES treatment affected basal cell development. In controls, basal cells were first evident at day 10 (vas deferens) or day 18 (cauda). Rats treated with 10 microg, but not those treated with 0.1 microg, DES, showed approximately 90% reduction (P<0.001) in basal cell numbers at day 15 and day 18. This decrease coincided with gross suppression of testosterone levels; co-treatment of rats with 10 microg DES+testosterone maintained basal cell numbers at control levels at day 18. However, suppression of testosterone production (GnRH antagonist treatment) or action (flutamide treatment) did not alter basal cell numbers. It is concluded that neonatal exposure to high oestrogen levels coincident with reduced testosterone action results in abnormal changes in the adult cauda/vas deferens that are preceded by delayed differentiation of basal cells. These findings imply a role for androgens and oestrogens in basal cell development and suggest that this may be pivotal in determining normal epithelial (and stromal) development of the cauda/vas deferens.     

Structure‐Functionality Changes in Starch Following Rough Rice Storage

The molecular‐level features of starch in relation to the changes in rice functionality during storage are not yet fully elucidated. This work investigated the effects of rough rice storage conditions on starch fine structures and physicochemical properties. Dried rough rice samples (medium‐grain Bengal and long‐grain Cypress) were stored at 4, 21, and 38°C in temperature‐controlled chambers and then periodically removed and evaluated after 1, 3, 5, 7, and 9 months. Flour (powdered head rice) and starch (extracted from head rice by alkali steeping) samples were evaluated for pasting and thermal properties. High‐performance size‐exclusion chromatography and high‐performance anion exchange chromatography were used to characterize starch molecular size and amylopectin chain‐length distribution, respectively. Significant changes in starch fine structure were observed primarily on the 38°C lots, and to some extent on the 21°C lots. The decreased amylose: amylopectin ratio, shortened amylopectin average chain length, and the shift in chain‐length distribution to shorter branch chains were implicative of molecular‐level starch degradation. The flour and starch samples showed inconsistent trends in pasting and thermal properties, thus suggesting the role of not only starch but also its interaction with non‐starch components in rice aging.     

Characterization of Physical Properties of Flour and Starch Obtained from Gamma‐Irradiated White Rice

Physical and structural characteristics of rice flour and starch obtained from gamma‐irradiated white rice were determined. Pasting viscosities of the rice flour and starch, analyzed by using a Rapid Visco Analyser, decreased continuously with the increase in irradiation dosage. Differential scanning calorimetry showed that gelatinization onset, peak and conclusion temperatures of rice flour and starch changed slightly but the enthalpy change decreased significantly with increase of irradiation dosage. All irradiated starch displayed an A‐type X‐ray diffraction pattern like the native starch. Gel permeation chromatography showed that the blue value ratio of the first peak (amylopectin) to the second one (amylose) decreased with the increase of the irradiation dosage. The weight‐average molecular weight (M_w) and gyration radius (R_z) of amylopectin analyzed by using HPSEC‐MALLS‐RI (high‐performance size‐exclusion chromatography equipped with multiangle laser‐light scattering and refractive index detector) decreased gradually from 1.48×10⁹ (M_w) and 384.1 nm (R_z) of native rice starch to 2.36×10⁸ (M_w) and 236.8 nm of 9 kGy‐irradiated starch. The branch chain‐length distribution of amylopectins determined by HPAEC‐ENZ‐PAD (high‐performance anion‐exchange chromatography with amyloglucosidase post‐column on‐line reactor and pulsed amperometric detector) showed that gamma irradiation had no significant effect on the amylopectin branch chains with 13≤DP≤24 and 37≤DP, but produced more branch chains with 6≤DP≤12 when the irradiation dosage was less than 9 kGy. It might be deduced that gamma irradiation caused the breakage of the amylopectin chains at the amorphous regions, but had little effects on the crystalline regions of starch granules, especially at low dosage irradiation.     

The relative importance of fungi in the breakdown of commercial samples of sulphited strawberries

The incidence of strawberry spoilage fungi in commercal plantations and on commercially harvested fruit from three different strawberry growing areas (East Norfolk, Wisbech area, County Wexford) were studied in relation to breakdown of sulphited fruit from these areas. Botrytis cinerea and Mucor piriformis were present on fruit from most plantations in the three areas whereas Rhizopus species were more prevalent on fruit from the Wisbech area. A higher proportion of stored fruit from the Wisbech area was spoiled by the Rhizopus spp., especially R. sexualls, whereas M. piriformis predominated on fruit from East Norfolk and County Wexford. A greater proportion of the sulphited fruit from the Wisbech area showed softening and disintegration which coincided with both a higher incidence of R. sexualis and of sulphited liquors showing detectable pectolytic activity. Comparison of the effect of the pre-harvest fungicides Elvaron, Mildothane and Daconil, showed a similar incidence of M. piriformis and Rhizopus spp. on the fruit and subsequently there were no consistent differences in the amount of breakdown of sulphited fruit from the different treatments. A delay in sulphiting after the fruit had been harvested, especially when fruit was held at 15°C, usually resulted in complete disintegration during subsequent storage. The importance of reducing the interval between harvesting and sulphiting to a minimum is discussed together with other factors which may reduce the incidence of breakdown of sulphited fruit.     

Reductive ethylation in ethereal solvents

Organic compounds, in particular anthracene and naphthalene, and two coals have been reductively ethylated in tetrahydrofuran (THF) and diglyme using alkali metals to form anions and ethyl iodide to produce ethylates. Product distributions were sensitive to experimental conditions. The rate of anion formation was controlled by their rate of desorption, probably as contact ion pairs, from the alkali metal surface. The enthalpy of formation of a solvated contact ion pair was ≈500 kJ mole^?1 mainly due to dipole-solvent interaction. 18-crown-6 ether was used to diagnose that, when treated with potassium in THF, anthracepe gave solvated contact ion pairs and probably di-anions whose reactions were characterized by the formation of ethyldihydroanthracene. Naphthalene under identical conditions gave loosely solvated mono-anions which reacted by electron transfer to give diethyldihydronaphthalene. With other alkali metals anthracene and naphthalene gave similar product distributions. The behaviour of other polynuclear aromatic hydrocarbons and of two coals was classified in the same way. It is suggested that when coals are readily solubilized by reductive alkylation mono-alkyldihydro products will predominate.     

Cholesterol blocks the disordering effects of ethanol in biomembranes

To assess the relation between the physical order of a membrane and its sensitivity to ethanol, we enriched biomembranes with cholesterol, both in vivo and in vitro. Japanese quail of the SEA line (selectively bred for susceptibility to experimental atherosclerosis) were treated for 9 to 16 weeks with a diet that contained 2% cholesterol. This regimen increased the cholesterol content of serum and erythrocytes. The cholesterol content of brain synaptosomal plasma membranes (SPM) was unaffected by the high cholesterol diet. In other experiments, isolated mouse synaptosomal plasma membranes were incubated with cholesterol/phospholipid (C/P) vesicles; different amounts of cholesterol were transferred according to the sterol content of the donor vesicles. Membrane order was determined in both types of membranes by a sensitive electron paramagnetic resonance (EPR) technique. The order parameter with 5- and 12-doxylstearic acid increased along with the cholesterol content. As expected, ethanol disordered membranes (decreased the order parameter) in a concentration-related manner. The slope of the concentration response curve was less steep in high cholesterol than low cholesterol membranes, indicating that cholesterol enrichment partially blocks the membrane action of ethanol in both types of membranes.     

Structures and Physicochemical Properties of Acid‐Thinned Corn,Potato and Rice Starches

The structures and physicochemical properties of acid‐thinned corn, potato, and rice starches were investigated. Corn, potato, and rice starches were hydrolyzed with 0.14 N hydrochloric acid at 50 °C until reaching a target pasting peak of 200—300 Brabender Units (BU) at 10% solids in the Brabender Visco Amylograph. After acid modification the amylose content decreased slightly and all starches retained their native crystallinity pattern. Acid primarily attacked the amorphous regions within the starch granule and both amylose and amylopectin were hydrolyzed simultaneously by acid. Acid modification decreased the longer chain fraction and increased the shorter chain fraction of corn and rice starches but increased the longer chain fraction and decreased the shorter chain fraction of potato starch, as measured by high‐performance size‐exclusion chromatography. Acid‐thinned potato starches produced much firmer gels than did acid‐thinned corn and rice starches, possibly due to potato starch's relatively higher percentage of long branch chains (degree of polymerization 13—24) in amylopectin. The short‐term development of gel structure by acid‐thinned starches was dependent on amylose content, whereas the long‐term gel strength appeared dependend on the long branch chains in amylopectin.