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[Do preparation techniques induce ultrastructural sperm damage? An electron microscopic study].

We investigated the possibility of ultrastructural damage to human spermatozoa induced by different sperm preparation techniques. Ejaculates from 20 normozoospermic men were divided into equal aliquots and processed by glass wool filtration, Percoll density gradient centrifugation, and a simple two-step centrifugation procedure which served as a control. Ultrastructural damage was assessed by scanning electron microscopy. Compared with Percoll density gradient centrifugation, glass wool filtration yielded a significantly increased proportion of intact acrosomes (p < 0.05). However, both preparations gave significantly better results than control (p < 0.001 and p < 0.01 for glass wool filtration and Percoll centrifugation, respectively). The significance of the conspicuous bending of sperm tails after Percoll centrifugation is not yet known. In conclusion, both glass wool filtration and Percoll centrifugation are efficient techniques for the accumulation of spermatozoa with intact acrosomes. Because of the higher sperm recovery rate, glass wool filtration appears to be the more appropriate method.

Acrosome↗

Differentiation of ejaculates showing reactive oxygen species production by spermatozoa or leukocytes.

Differences between subgroups and correlations between reactive oxygen species (ROS), sperm motility, concentration of leukocytes and viability in semen samples from 143 men were investigated. Patients with azoospermia or leukocytospermia were excluded from the study. Spermatozoa were separated by means of glass wool filtration. Reactive oxygen species were determined by means of luminol chemiluminescence before and after sperm separation; thereafter, normozoospermic and oligozoospermic patients were divided into three subgroups using the mean of all patients investigated (17462 count 10(-7) viable spermatozoa) as cut-off value as follows: G1--high reactive oxygen species production in native semen and after glass wool filtration; G2--high production of reactive oxygen species only in native semen; G3--low levels of reactive oxygen species before and after glass wool filtration. In general, reactive oxygen species were significantly higher in oligozoospermic samples than in normozoospermic samples. In men with normal sperm count, ROS production correlated significantly with the number of leukocytes in the ejaculate before glass wool filtration, but not thereafter. Glass wool filtration is useful to distinguish between sperm samples in which reactive oxygen species are generated by leukocytes and those in which reactive oxygen species are excessively generated by the spermatozoa themselves.

Humans↗

Cat allergen sampling by a new personal collector (Partrap FA 52).

Recent studies carried out by us and others have demonstrated that Fel d 1, the main cat allergen, may be passively transferred by human clothing in cat-free environments. Consequently, the monitoring of the Fel d 1 levels either in indoor environments or on allergen-contaminated clothes of sensitized cat owners should be considered an important tool in prevention strategies. The aim of this study was to evaluate the efficacy of a personal air sampler (Partrap FA 52) in capturing cat allergen from wool fabrics. Seven identical wool webs (80 x 100 cm) were put in the baskets of seven male cats for 1 week. In our laboratory each web was divided into two parts (80 x 50 cm), the first of which was then divided in two parts (40 x 50 cm) and each was vacuumed directly by one collector. The second part was dry-cleaned at a professional cleaners, divided in two parts and then vacuumed. For the dust collection from wool webs we used a fixed high volume air sampler (CF/20 Gelaire Flow Labs, Milan, Italy) and a personal collector (Partrap FA 52, Coppa Biella, Italy). Fel d 1 content was determined using a two site ELISA (ALK-Abelló Group, Madrid, Spain). Both air samplers collected cat allergens from cat-exposed wool fabrics before and after dry cleaning. There were significant differences between the levels of Fel d 1 before and after dry cleaning by using either CF/20 or Partrap FA52 and between the levels of Fel d 1 before dry cleaning using CF/20 and Partrap FA 52. The results of our study suggest that Partrap FA 52, although its air flow is half that of the CF/20, is able to collect even residual amounts of cat allergen from wool webs after dry cleaning and consequently may constitute a simple and effective means of monitoring the levels of Fel d 1 on the clothes of cat owners.

Air Pollution, Indoor↗

Monofilament assessment of neuropathy in a community diabetes clinic.

OBJECTIVE: To compare the detection of diabetic neuropathy using monofilament, cotton wool, pinprick, vibration sense and symptom evaluation. SETTING: The diabetes clinic of a community hospital. METHODS: Two examiners evaluated 89 women with diabetes mellitus (DM) using a 10 g monofilament, cotton wool, pinprick and a 128 Hz tuning fork after completion of a University of Texas subjective peripheral neuropathy verbal questionnaire. RESULTS: Vibration sense was abnormal in either foot in 8% of subjects. Neuropathy as defined by monofilament, cotton wool and pinprick was present in 26%, 3% and 6% of patients respectively. The respective kappa values (kappa) for the comparison between monofilament neuropathy and cotton wool neuropathy, pinprick neuropathy and symptom-defined neuropathy were 0.18, 0.21 and 0.06. The kappa-value comparing monofilament and tuning fork-defined neuropathy was 0.24. There was fair agreement between 10 versus 3 sites (kappa = 0.60). CONCLUSION: More abnormalities were detected using the monofilament compared with cotton wool or pinprick. There was poor concordance between symptoms and clinically detected neuropathy. The ideal number of sites that need to be evaluated is still contentious.

Cross-Sectional Studies↗

CD4+ suppressor cells differentially affect the production of IFN-gamma by effector cells of experimental autoimmune encephalomyelitis.

Spleen cells from rats that have recovered from experimental autoimmune encephalomyelitis (EAE) suppress the production of IFN-gamma by effector T cells of EAE in an Ag-specific manner. These postrecovery suppressor cells also inhibit EAE in vivo. Fractionation of the postrecovery suppressor spleen cells on nylon wool and OX-8 coated plates yields a nylon wool-adherent CD4+ suppressor cell population that, when cocultured with effector T cells, suppresses IFN-gamma production by these effector cells. In contrast, the nylon wool-adherent, CD4+ postrecovery suppressor cell population fails to inhibit the production of IL-2 by the effector T cells. In further experiments, the effector T cell population was depleted of CD8+ cells and cocultured with the nylon wool-adherent, CD4+ postrecovery suppressor cells, and the supernatants were assayed for IFN-gamma and IL-2. IFN-gamma production was inhibited in these cultures but IL-2 production was not inhibited. Irradiated effector T cells were cocultured with CD4+ postrecovery suppressor cells, without myelin basic protein, in an effort to determine whether the mechanism of differential lymphokine suppression involved an anti-idiotypic response against effector T cells. No IL-2 was produced, indicating that there was no CD4+ suppressor cell mediated anti-idiotypic response against effector T cells. These studies suggest that the suppressor cell is a nylon wool adherent, CD4+ T cell that functions to down-regulate EAE effector T cells by differential inhibition of lymphokine production.

Animals↗

Environmental surveys in the European man-made mineral fiber production industry.

This paper presents estimates of airborne fiber concentrations and fiber size for European man-made mineral fiber (MMMF) factories on the basis of measurements made in 1977-1980. The airborne fiber concentrations previously reported at a conference of the World Health Organization (WHO) and the International Agency for Research on Cancer in 1982 have been revised to harmonize the results with the WHO-European MMMF reference counting level. The result was an approximate doubling of the reported airborne fiber levels. After the revisions the average combined occupational group concentrations in the rock- and glass-wool plants were still generally low (less than 0.01 fibers/ml). In the glass continuous-filament factories the airborne fiber concentrations were very low (less than 0.01 fibers/ml). The average plant median for fiber length ranged from 10 to 20 microns, and the corresponding median diameters ranged from 0.7 to 2 microns. In general the glass-wool fibers were thinner than the rock-wool fibers. The fiber concentrations measured in other studies in the MMMF production and user industries are reviewed. Higher levels (between 0.1 and 1.0 fibers/ml) have been measured in some insulation wool production, secondary production, and user industries. The highest levels (greater than 1.0 fibers/ml) occurred in very fine glass-fiber production and in other specialist insulation wool usage.

Air Pollutants, Occupational↗

[Electron microscopic analysis of possible ultrastructural damage to spermatozoa after preparation by various methods].

We investigated the possibility of ultrastructural damage to human spermatozoa induced by different sperm preparation techniques. Ejaculates from 20 normozoospermic men were divided into equal aliquots and processed by glass wool filtration, Percoll density gradient centrifugation, and a simple two step centrifugation procedure which served as a control. Ultrastructural damage was assessed by scanning electron microscopy. Compared with Percoll density gradient centrifugation, glass wool filtration yielded a significantly increased proportion of intact acrosomes (p < 0.05). However, both preparations gave significantly better results than control (p < 0.001 and p < 0.01 for glass wool filtration and Percoll centrifugation, respectively). The conspicuous bending of sperm tails after Percoll centrifugation has a yet unknown importance. In conclusion, both glass wool filtration and Percoll centrifugation are efficient techniques for the accumulation of spermatozoa with intact acrosomes. Because of the higher sperm recovery rate, glass wool filtration appears to be the more appropriate method.

Acrosome↗

DNA profiling of shahtoosh.

The population of the Tibetan Antelope (Pantholops hodgsonii) has recently declined dramatically due to the illegal trade in its wool. The animal lives at high altitude and is protected from the extremely cold climate due to a coat of very fine wool. These hairs are highly sought for weaving a shawl called shahtoosh. The large-scale poaching of the antelope has resulted in the species being placed on CITES Appendix I. We report on a method of DNA profiling on a confiscated shahtoosh using the cytochrome b (cyt b) gene to produce a test that will identify unambiguously the presence of P. hodgsonii. Two shahtoosh samples were provided by the Council of Agriculture, Taiwan, and ten shawl samples of sheep wool (Ovis aries), cashmere from the Kashmir goat (Capra hircus), and pashmina from the Himalayan goat (C. hircus) were collected from local stores for comparison. Two primer pairs were used to amplify a 271 bp fragment of cyt b gene which would distinguish these three species. The resulting amplification products were directly sequenced. When the DNA sequences were compared with the sequences registered in GenBank and EMBL databases, the sequences with the highest homology were the cyt b genes of P. hodgsonii, C. hircus, and O. aries. This study demonstrates that there is still sufficient DNA present in the finished wool of a shahtoosh shawl to allow DNA typing and the method established was highly plausible to identify the CITES protected species.

Animals↗

Restricting detergent protease action to surface of protein fibres by chemical modification.

Due to their excellent properties, such as thermostability, activity over a broad range of pH and efficient stain removal, proteases from Bacillus sp. are commonly used in the textile industry including industrial processes and laundry and represent one of the most important groups of enzymes. However, due to the action of proteases, severe damage on natural protein fibres such as silk and wool result after washing with detergents containing proteases. To include the benefits of proteases in a wool fibre friendly detergent formulation, the soluble polymer polyethylene glycol (PEG) was covalently attached to a protease from Bacillus licheniformis. In contrast to activation of PEG with cyanuric chloride (50%) activation with 1,1'-carbonyldiimidazole (CDI) lead to activity recovery above 90%. With these modified enzymes, hydrolytic attack on wool fibres could be successfully prevented up to 95% compared to the native enzymes. Colour difference (DeltaE) measured in the three dimensional colour space showed good stain removal properties for the modified enzymes. Furthermore, half-life of the modified enzymes in buffers and commercial detergents solutions was nearly twice as high as those of the non-modified enzymes with values of up to 63 min. Out of the different modified proteases especially the B. licheniformis protease with the 2.0-kDa polymer attached both retained stain removal properties and did not hydrolyse/damage wool fibres.

Animals↗

Metabolic response of sheep skin to a chronic infusion of a variant of insulin-like growth factor I.

The effects of a chronic (21-day) skin infusion of a variant of insulin-like growth factor I (IGF-I) (long-Arg3-IGF-I; LR3IGF-I) on short-term (48 h) responses of skin metabolism and 21-day plasma hormone concentration, wool-follicle characteristics and wool production were investigated in well-fed castrated Romney sheep. A bilateral arteriovenous preparation was used to infuse LR3IGF-I continuously into the skin on one abdominal flank and saline into the other abdominal flank of six sheep; a further six sheep had one flank infused with saline (controls). LR3IGF-I caused an initial (4-24 h) reduction in the plasma concentrations of amino acids, especially tyrosine, valine and lysine, and, after 24 h, significant (P < 0.05) reductions in blood oxygen and plasma glucose concentrations. After 4 h of LR3IGF-I infusion, there was a significant increase in blood flow (P < 0.05) and oxygen uptake (P < 0.05), and net uptake of amino acids [which was significant (P < 0.05) for valine and phenylalanine] by the LR3IGF-I-infused skin was increased. Total uptake of phenylalanine for skin protein synthesis, measured using [3H]phenylalanine uptake, was also significantly increased after 4 and 24 h of infusion. After 48 h of infusion all LR3IGF-I-dependent measurements of metabolic parameters had fallen to preinfusion values. By day 7 of the 21-day infusion there was a significant (P < 0.05) decrease in circulating endogenous IGF-I in plasma of treated sheep compared with that of control sheep, followed by a significant (P < 0.05) increase between day 7 and 21. Plasma insulin levels followed a similar pattern. There was no change at any stage of infusion in IGF-binding proteins in the plasma of the two LR3IGF-I-infused sheep tested, and it is concluded that LR3IGF-I caused a down-regulation of the type-I IGF-I receptors followed by a rise in endogenous IGF-I concentration consequent on lack of feedback regulation. After 21 days of infusion there was no effect of LR3IGF-I on wool-follicle-bulb-cell mitotic rate, bulb diameter or wool production.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗

Filtration characteristics of three new in-line blood transfusion filters.

Three types of microemboli filters were examined with the scanning electron microscope. The single layer 40 mu pore filter traps aggregates in the pores. The aggregates are extruded and pass on with the blood and can be demonstrated in a Dacron wool filter downstream. The polyurethane foam filter is an in depth multiple pore filter of variable pore size. This filter allows many aggregates to escape through the pores in the beginning of a transfusion and these aggregates were seen in a Dacron wool filter downstream. As aggregates become trapped, the pore sizes decrease, filtration improves, and flow through the filter becomes reduced. The Dacron wool filter absorbs the particulate material to the fibers of wool. This filter has a large capacity and was not observed to leak aggregates even when subjected to high filtration pressures.

Animals↗

The prevalence of lice (Bovicola ovis) infested sheep flocks in Western Australia (1987-1993)

The proportion of wool bale brands with a positive test for sheep lice in baled wool decreased from 29.5% in 1987/88 to 23.2% in 1990/91 before increasing to 38.2% in 1992/93. Changes in the proportion of wool bale brands with a positive test for lice were highly correlated with changes in the Wool Market Price Indicator. The increase in the proportion of positive lice tests since 1990/91 was associated with an increase in failures to eradicate lice from flocks. These failures were partly a consequence of the reduced use of lousicidal treatments, the development of resistance to synthetic pyrethroid chemicals and an increase in the transmission of lice between flocks.

Animals↗

Persistence of Salmonella typhimurium on fabrics.

The persistence of Salmonella typhimurium (V-31) on wool blanket, wool gabardine, cotton sheeting, cotton knit jersey, cotton terry cloth, and cotton wash-and-wear fabrics was studied. Three methods of exposure were employed to contaminate the fabrics: direct contact, aerosol, and a lyophilized mixture of bacteria and dust having a high content of textile fibers. After contamination, the fabrics were held in 35 or 78% relative humidity at 25 C. The persistence time of S. typhimurium on fabrics held in 35% relative humidity was substantially longer when the fabrics were contaminated by direct contact or by exposure to dust containing bacteria than when contaminated by exposure to aerosolized cultures. Viable bacterial populations persisted for 24 weeks at relatively high population densities on swatches of wool gabardine, cotton sheeting, cotton knit jersey, and cotton terry cloth exposed by direct contact and held in a humidity of 35%. In 78% humidity, bacterial populations persisted on the fabrics for relatively shorter periods of time regardless of the mode of contamination or fabric type. This organism retained its virulence for Swiss mice after being recovered from wool gabardine swatches held 8 weeks in humidities of 35 or 78% and from cotton terry cloth swatches held 6 weeks in the same humidities.

Aerosols↗

LFA-1 and ICAM-1 in homotypic aggregation of rat alveolar macrophages: organic dust-mediated aggregation by a non-protein kinase C-dependent pathway.

We have examined the role of cell adhesion molecules in the homotypic aggregation of rat alveolar macrophages after exposure to wool and grain dusts. Molecules such as bacterial lipopolysaccharide (LPS) and phorbol myristate acetate (PMA) can upregulate adhesion molecules, resulting in aggregation of lymphocytes. In rats treated intratracheally with an inspirable sample of wool dust collected from the air of British wool textile mills, and sieved grain dust, aggregates of macrophages were present in the bronchoalveolar lavage (BAL). Our hypothesis was that substances present on the dust surface could activate and upregulate adhesion molecules of the CD11/CD18 complex on the BAL cells and account for the aggregates. Macrophages from untreated rats form aggregates in vitro, which averaged 19 cells/aggregate; when treated with both wool and grain dusts, this rose to 25 and 24 cells/aggregate, respectively. LPS, PMA, and the proinflammatory cytokine tumor necrosis factor (TNF) also caused increases in aggregate size. Staurosporine, an inhibitor of protein kinase C (PKC), reduced the number of cells per aggregate from 35 cells/aggregate in LPS- and PMA-treated macrophages to 18 cells/aggregate, the same as untreated. In contrast, staurosporine had no effect in reducing the size of aggregates produced by the organic dusts. Removal of divalent cations, which are essential for maintaining integrin stability and PKC activity, resulted in complete abolition of aggregate formation. Treatment with monoclonal antibodies to lymphocyte function-associated antigen-1 (LFA-1) alpha and beta and intercellular adhesion molecule-1 (ICAM-1) resulted in the inhibition of aggregate formation in a dose-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaloids↗

Chronic vascular reactions to steel coil occlusion devices.

Original Gianturco steel coil vascular embolization devices and two modifications were compared for their occlusive effects and for the degree of adverse reaction in the surrounding vessel wall. Modifications consisted of silk or Ivalon replacing the wool strands of the original device. Single coils were placed into the left renal artery of nine mixed breed swine and restudied 3 months later. The bare coils did not occlude the vessels nor lead to significant foreign body reaction. Wool, silk, and Ivalon coils produced prompt and sustained renal artery occlusion. Upon histologic examination of the vessel wall, foreign body and chronic inflammatory reaction was found to be strongest against wool. This reaction was less with silk and markedly diminished with Ivalon. The results indicate that silk and Ivalon coils, while having the same occlusive effect, are preferable to wool coils if less local reaction is desired.

Alloys↗

Tolerance of inorganic selenium by range-type ewes during gestation and lactation.

The objectives of this 72-wk study were to evaluate and compare the effects of 6 dietary levels of inorganic Se on serum, whole blood, wool, and tissue Se concentrations and to determine the maximum tolerable level of Se for mature ewes during lamb production. Forty-one, 4-yr-old, range-type ewes (57.4 +/- 5.7 kg) were used in a completely randomized design with 6 dietary treatments. Sodium selenite was added to a corn and soybean meal-based diet to provide 0.2 (control), 4, 8, 12, 16, or 20 mg of dietary Se/kg to ewes during lamb production. Serum Se and ewe BW were measured at 4-wk intervals; whole blood Se and wool Se were measured every 12 wk; and samples of brain, diaphragm, heart, hoof, kidney, liver, and psoas major were collected at the termination of the experiment. Dietary Se did not affect ewe BW during the study (P = 0.69), and there was no treatment x time interaction. Serum Se increased linearly as dietary Se level increased (P < 0.001) and responded cubically (P = 0.02) over time. Selenium in whole blood increased linearly (P < 0.001) as supplemental Se increased. Wool Se increased linearly (P < 0.001) as dietary Se increased, and the response over time was quadratic (P < 0.001). Brain, diaphragm, heart, and psoas major Se increased (P < 0.05) linearly as dietary Se increased, liver Se responded quadratically (P < 0.05), and hoof and kidney Se increased cubicically (P < 0.05) as supplemental Se increased. In general, serum, whole blood, and tissue Se concentrations of ewes receiving 12, 16, or 20 mg of dietary Se/kg were greater (P < 0.05) than those of controls and ewes receiving less dietary Se. Although they were elevated in ewes receiving increased dietary Se, at no time did serum, whole blood, or wool Se concentrations reach levels previously reported as toxic, nor were clinical signs of Se toxicosis observed. Histopathological evaluation of liver, kidney, diaphragm, heart, and psoas major did not reveal evidence of Se toxicosis in ewes at any dietary Se level. Ewes under our experimental conditions and during the stresses of production were able to tolerate up to 20 mg of dietary Se/kg as sodium selenite for 72 wk. These findings suggest that the maximum tolerable level of inorganic Se for sheep is much greater than 2 mg/kg as was suggested previously. Experiments of longer duration and utilizing greater dietary Se concentrations are necessary to clearly define the maximum tolerable level.

Animal Feed↗

Effect of anthelmintic treatment on the productivity of lambs infected with the intestinal nematode, Trichostrongylus colubriformis.

Liveweight gain, food intake, wool growth and concentration of serum constituents were measured in four groups of three-month-old lambs. Groups 1 and 2 were infected on five days each week with 2500 Trichostrongylus colubriformis larvae for 18 weeks. Group 2 lambs were treated with fenbendazole (5 mg kg-1) at week 10 and week 15. Group 3 lambs were similarly infected for 10 weeks, treated with fenbendazole, but given no further larvae. A fourth control group remained uninfected throughout the trial. All lambs were killed at week 20. Mean worm populations were 16,130, 430 and zero for groups 1, 2 and 3 respectively. Over the first 10 weeks of infection, liveweight gain was reduced by 52, 43 and 51 per cent and wool growth by 33, 28 and 27 per cent respectively, in groups 1, 2 and 3. Serum hypophosphataemia and hypoalbuminaemia occurred in all three infected groups. Between weeks 10 and 20 overall weight gain and wool growth in group 1 lambs were 44 and 46 per cent lower than the controls, whereas weight gains of group 2 and group 3 lambs were similar to or slightly higher than in the controls. However, wool growth in these two groups after treatment was only 73 to 74 per cent of control values. Serum phosphorus concentrations increased to control levels within one week of anthelmintic treatment and serum albumin concentrations by two to four weeks. The ability of group 2 lambs to improve their performance after anthelmintic treatment, in the face of continued challenge, was attributed to development of resistance to reinfection.

Animals↗

Cytotoxicity to tumors by alpha, beta-dihydric long-chain fatty alcohols isolated from esterolysates of uncytotoxic sheep cutaneous wax: the dependence on the molecular hydrophobicity balance of N- or iso-alkyl moiety bulkiness and two hydroxyl groups.

Wool fatty alcohols (WF-Alc; C10-C33), separated by esterolysis of wool grease secreted from sheep sebaceous gland, inhibited growth of mouse Ehrlich ascites carcinoma (EAC) cells in contrast to no inhibition by unesterolysed wool grease. WF-Alc was fractionated by molecular distillation and subsequent octadecylsilica (ODS) gel liquid chromatography, showing that most of the growth-inhibitory activity was found in the most hydrophilic fraction with the lowest boiling-point (MW 200-300; C12-C20), ODS-HPLC of the fraction showed that most of the activity resided in two homogeneous fractions identified by GC-MS and 13C-/1H-NMR as alpha, beta-dihydric saturated fatty alcohols such as 1,2-hexadecanediol (n-C16(OH)2) and 16-methyl-1,2-heptadecanediol (iso-C18(OH)2), respectively. EAC cells implanted into mice were inhibited markedly by n-C16(OH)2 possessing a cytolysing ability and slightly by iso-C18(OH)2, but hardly by other alkyl-alpha, beta-diols (C12-C24) contained in WF-Alc. Thus, the antitumor activity of WF-Alc was exhibited only after saponification of uncytotoxic wool grease, showing necessity of unesterified hydroxyl groups, and was dependent upon the molecular hydrophobicity balance attributed to both hydroxyl groups and n- or iso-alkyl moiety bulkiness specified out of diverse species of fatty alcohols contained in WF-Alc.

Animals↗