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The combination of two semen preparation techniques (glass wool filtration and swim-up) and their effect on the morphology of recovered spermatozoa and outcome of IVF-ET.

The aim of this prospective study was to relate sperm quality, especially sperm morphology, to the outcome of IVF results, when glass wool filtration (GWF) and swim-up were used in combination for the preparation of spermatozoa. A total of 60 ejaculates were analysed. GWF was compared with glass wool filtration/swim-up (GWF-SU) by using aliquots from the same semen samples to increase the precision of the comparison and to establish the cumulative effect of these two semen preparation methods on the morphology of the recovered spermatozoa. Sperm parameters were examined in native semen, in semen preparation samples after GWF and GWF-SU. The mean percentages for motility, morphology and velocity were improved significantly over those in fresh semen only when GWF-SU was used. GWF alone resulted only in a significantly higher recovery of motile spermatozoa. In semen preparation, 10% of spermatozoa with normal morphology appeared to be the cut-off point as there was a significantly higher recovery in fresh semen samples containing > 10% morphologically normal spermatozoa after GWF-SU. Outcome of IVF-ET following preparation with GWF-SU showed better results in comparison to the swim-up procedure alone, though this difference was not statistically significant. The cut-off point was also > 10% morphologically normal spermatozoa. Improvement in all aspects of IVF-ET occurred when native semen contained > 10% morphologically normal spermatozoa. It is concluded that GWF alone did not produce significantly better results but, in combination with swim-up, it resulted in significantly better sperm morphology and in an improved outcome of IVF-ET for fresh semen samples with > 10% morphologically normal spermatozoa.

Adult↗

2-Mercaptoethanol-dependent lipopolysaccharide-responsive B cells in nylon-wool-fractionated spleen cell preparations.

The ability of lipopolysaccharide (LPS) to influence lymphocyte mitogen responsiveness to 2-mercaptoethanol (2-ME) was investigated. 2-ME was mitogenic for splenocytes only when fetal bovine serum lots containing elevated endotoxin levels were used to supplement (5%) the culture medium. 2-ME did not stimulate mitosis of nylon-wool-fractionated splenocytes (NWSC); however, 2-ME induced significant proliferation in the NWSC cell preparations when endotoxin-deficient culture medium was supplemented with doses of LPS which alone did not stimulate significant proliferation. The sIg+ and Thy-1-/Lyt-1- fractions of NWSC, which represented less than 5% of the cells in the 'T-cell preparations', were almost completely dependent on 2-ME for proliferation in response to LPS stimulation. These data indicate that nylon-wool-fractionated preparations contain splenic B cells that incorporate significant amounts of [3H]thymidine when stimulated with LPS and 2-ME.

Animals↗

Improved B cell typing for HLA-DR using nylon wool column enriched B lymphocyte preparations.

A rapid, simple procedure for preparing B lymphocytes using nylon wool has been devised. When mixtures of T lymphocytes, B lymphocytes, and monocytes are applied to nylon wool columns, B lymphocytes can then be removed, virtually free of monocytes. Such preparations (Ad) are 85 +/- 6% SIg+ and have only 3 +/- 1% monocyte contamination. The yield from peripheral blood averages 5% of all mononuclear cells. In contrast, the E rosette depletion method (E-) yields cells which are only 64 +/- 5% SIg+ and have 25 +/- 7% monocyte contamination. The superiority of the nylon method for HLA-DR typing was demonstrated in a comparative study of Ad and E- with eight normal individuals. Cytotoxic scores were higher and a large number of reactions, representing DRw groups and additional cross-reactions, was detected.

B-Lymphocytes↗

In vitro and in vivo evaluation of platelet concentrates after cotton wool filtration.

Removal of leukocytes from platelet concentrates (PC) may decrease the incidence of alloimmunization to HLA antigens on white cells and prevent or delay refractoriness. Cotton wool filtration, which effectively removes leukocytes from PC with minimal platelet loss, may cause platelet or complement activation. In this study, the effect of cotton wool filtration (Imugard IG-500) on platelet activation, in vitro function, posttransfusion survival, and complement activation was investigated. Five paired autologous in vivo percent recovery and survival studies were performed with fresh (6-8h) PC or with 5-day-stored PC prepared from CPD-anticoagulated blood and stored in PL-732(TM) containers using standard methods. In the paired design, PC from the same donor were filtered on one occasion, and not filtered on another, prior to labeling with indium-III-oxine and reinfusion. Percent recovery and survival were then determined by the standardized method. Filtration had no statistically significant effect on percent recovery on PC stored for 6-8h or for 5 days. There was, however, a slight decrease in survival hours of the filtered PC which was statistically significant at 6-8h (203 +/- 14 vs. 179 +/- 18h; p less than 0.05) but not at 5 days (166 +/- 28 vs. 132 +/- 27h; p greater than 0.05). Samples taken before and after filtration of single units (fresh and 5-day) and pooled units (3-day) for measurement of release of granular content (beta-thromboglobulin), lysis (LDH), and in vitro viability - ATP, extent of shape change with ADP, and hypotonic shock response (3-day only)-demonstrated no effect of filtration on these parameters.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Glass wool filter preparation of cryopreserved spermatozoa.

Oligozoospermic and asthenozoospermic semen ejaculates, as well as cryopreserved sperm samples prepared by the wash and swim-up procedure often result in unsatisfactory sperm recovery rates. In this study the glass wool filter and the wash and swim-up preparation procedures were compared on the basis of their 'effective' (number of live sperm per millilitre) recovery rates. The glass wool filter procedure consistently produced significantly (P = 0.0002) higher viable sperm concentrations, making it the preferred method for the preparation of cryopreserved sperm to be used in assisted reproduction techniques. The use of this preparation procedure has also been shown to have no adverse affect on the fertilizing potential of human spermatozoa in our unit.

Cryopreservation↗

Measurement of albumin and low molecular weight proteins in the urine of newborn infants using a cotton wool ball collection method.

OBJECTIVE: The aim of this study was to investigate the inter-relationship between urinary excretion of alpha-1-microglobulin (A1M), retinol-binding protein (RBP) and albumin in term and premature neonates, with urine collected into cotton wool balls and extracted by a novel method. SUBJECTS AND METHODS: Sixty-four infants were studied on the first day of life; 26 had been born at term (37-42 weeks gestation) and 38 prematurely (24-28 weeks n = 16, 29-36 weeks n = 22). Urine collected into cotton wool balls was analysed following a new detergent extraction method, which resulted in a recovery rate of 94-107% for albumin. A1M, RBP and creatinine. RESULTS: Urinary protein excretion, expressed as a ratio to urinary creatinine, decreased significantly with increasing gestational age (24-28 weeks, 29-36 weeks, 37-42 weeks: albumin:creatinine ratio mg/mmol mean 96.9, 31.7, 19.3; A1M:creatinine ratio mg/mmol mean 99.3, 37.0, 7.8; RBP:creatinine ratio mg/mmol mean 16.2, 3.8, and < 0.01, below the limit of detection, respectively). When results were corrected for birthweight, this gestation-associated effect was still present for A1M and RBP, but not for albumin. In premature infants there was a significant positive correlation between A1M:creatinine ratio and RBP:creatinine ratio (r = 0.85), and also between albumin and both A1M and RBP (r = 0.82 and 0.77). CONCLUSION: Increased excretion of A1M, RBP and albumin at earlier gestational ages is probably due to proximal tubular immaturity, although tubular damage and also glomerular dysfunction cannot be excluded as possible explanations.

Albuminuria↗

Why cotton wool spots should not be regarded as retinal nerve fibre layer infarcts.

Cotton wool spots (CWSs) comprise localised accumulations of axoplasmic debris within adjacent bundles of unmyelinated ganglion cell axons. Their formation is widely held to reflect focal ischaemia from terminal arteriolar occlusion, but credible evidence supporting this view is lacking. CWSs are here purported to be nothing more than sentinels of retinal nerve fibre layer pathology, hence their recommended redesignation "cotton wool sentinels." After branch arteriolar occlusion, CWSs evolve as boundary sentinels of infarction, their uniform width suggesting a glial constraint to axonal expansion. In pre-proliferative diabetic retinopathy, CWSs form a C-shaped chain nasal to the disc and around the macula where they constitute sentinels of ischaemia affecting the entire retinal mid-periphery. The polymorphous CWSs evolving during acute panretinal hypoperfusion represent sentinels of an ischaemic penumbra. Those surrounding the disc in Purtscher's traumatic angiopathy are sentinels of neuronal damage from transient venous hyperdistension that overwhelms the protection afforded by peripapillary axonal decompartmentalisation.

Axonal Transport↗

Atypical cotton-wool spots.

Atypical cotton-wool spots in four selected cases of diabetic retinopathy are discussed. The most striking characteristic distinguishing them from typical cotton-wool spots is their size, which ranges from about 2 to 4 disc diameters. They develop after stenosis or a complete obstruction of a first order arteriole at the point at which it emerges from the parent arterial branch. The size of the lesion corresponds with the size of the affected arteriole. Restoration of local circulation is attempted by neighbouring arterioles and venules, the latter carrying a reversed blood flow. Arterio-venous communications along the border of the infarcted area are a prominent feature, most likely attributable to the necessity of free drainage of the arterial influx. The same haemodynamic principle applies to the coarsening of the adjacent capillary bed. Secondary changes of the bordering venous branches include aneurysm formation and the staining of the venous wall.

Aged↗

Multiple cotton wool spots following bone marrow transplantation for treatment of acute lymphatic leukaemia.

Three patients with acute lymphatic leukaemia developed visual impairment due to occlusion of small retinal vessels with multiple cotton wool spots after treatment which included whole body and skull irradiation followed by bone marrow transplantation and cyclosporin A. Withdrawal of cyclosporin A and treatment with corticosteroids was followed by recovery of visual acuity. This retinopathy and the retinal changes seen in the immunodeficiency syndrome are thought to be closely related. The possible role of cyclosporin A is discussed, though cotton wool spots and retinal haemorrhages have never been described in renal transplant patients during treatment with this drug. Withdrawal of cyclosporin A, which is highly effective in preventing graft-versus-host disease, can be fatal. Irradiation of the skull prior to bone marrow transplantation and intrathecal administration of methotrexate may be the most important factors causing the retinal ischaemic signs described here. The inclusion of an ophthalmologist in the team monitoring transplant patients would lead to increased documentation and a better understanding of this disease.

Adult↗

Retinal cotton-wool spots: an early finding in diabetic retinopathy?

Five insulin dependent diabetic patients are reported on who had a few small retinal cotton-wool spots or 'soft exudates' either totally isolated or associated with fewer than 10 microaneurysms. These observations suggest that cotton-wool spots may be an early finding in diabetic retinopathy. Significant biological abnormalities in these patients were high levels of glycosylated haemoglobin and mild increases in thrombin generation, indicating slight activation of the coagulation system. The possible significance of these clinical and biological findings is discussed.

Adult↗

Survival of upper respiratory tract bacteria on cotton-wool swabs.

Plain, buffered cotton-wool swabs and albumen-coated cotton-wool swabs were prepared with known numbers of the following: Haemophilus influenzae, Streptococcus pneumoniae, Staphylococcus aureus, corynebacteria and Escherichia coli. Swabs were plated immediately (0 hours) after loading with a 0.1 ml standardised inoculum; each swab was plated out four times by streaking on both halves of two plates. At 0 hours recovery rates from first platings on solid media ranged from 3.3%-8.9%; from platings 1-4 ranges were 2.5%-5.7%. Both types of swabs gave similar results. The greater the delay in plating swabs at room temperature and 4 degrees C the lower was the recovery rate of H. influenzae and Strep. pneumoniae, though both species survived better on either kind of swab when these were held at 4 degrees C. There was a very small reduction in numbers of Staph. aureus from 24 to 48 hours but the recovery rate of corynebacteria was similar over the 48-hour period, at both room temperature and 4 degrees C. Twenty-four-hour counts of E. coli at room temperature were approximately five times greater than at 0 hours, and at 48 hours colonies were too numerous to count. At 4 degrees C, however, counts of E. coli fell during the 48-hour period.

Albumins↗

Single isolated cotton-wool spots.

The presence of isolated cotton-wool spots in normal fundi may indicate serious systemic disease. In this presentation 4 patients were examined for blurred vision or scotomas with sudden onset. Two of them were pregnant. Funduscopic examination revealed soft-appearing white spots in the temporal arcades in 3 patients and in the papillomacular bundle in 1 patient. Due to the lack of other fundus lesions 3 of the patients were initially treated for toxoplasmosis. The patients were seronegative for typical viral (HIV), antinuclear antibodies, bacterial infections, or toxoplasmosis. Blood pressures, glycosylated hemoglobin and blood counts were in the normal range. At the follow-up examinations subjective symptoms had improved and the spots disappeared or were smaller and showed more distinct borders, and no new lesions were detected. These cases suggest that isolated cotton-wool spots may occur without serious systemic disease in otherwise healthy subjects and the spots may be misinterpreted as acute inflammatory retinochoroidal lesions.

Adult↗

Subpopulations of pig blood E-rosette-forming lymphocytes and thymus-dependent null cells: separation by nylon wool columns, rosette formation and macrophage-dependent mitogen and antigen responsiveness.

Pig peripheral blood lymphocyte subpopulations were characterized by their adherence to nylon wool. Thus whilst B cells, defined by surface Ig and Fc receptor, were predominantly adherent, thymus-dependent Null cells and the T cells forming rosettes with sheep red blood cells in dextran (DS+) were predominantly non-adherent. There was a direct relationship between the size of the non-adherent fraction and the proportion of the total DS+ cells which were in the non-adherent fraction. Using nylon wool and/or DS rosette separation, lymphocytes from normal and BCG sensitized animals were stimulated in vitro with phytohemagglutinin (PHA), concanavalin A (Con A) and tuberculin (PPD). The cells responding to PHA and Con A (T cell mitogens in other species) were DS+ lymphocytes. Non-rosetting, adherent (B cells) and non-adherent (thymus dependent Null cells) lymphocytes were unresponsive, even in the presence of homologous alveolar macrophages at levels which greatly enhanced the PPD response. Cells responding to PPD were present in the DS+ and DS-, adherent and non-adherent fractions but their response was apparently controlled by macrophage dependence and suppression. Techniques are described for the preparation and storage in liquid nitrogen of alveolar macrophages resulting in high numbers of viable cells, capable of helping in vitro responses, even after 3 years of storage.

Animals↗

Extracorporeal glass-wool filtration of whole blood enhances post-ischemic recovery of the cortical sensory evoked response.

In twenty dogs, anticoagulated with heparin 300 units/kg, the right cortical sensory evoked response (CSER) to contralateral median nerve stimulation was suppressed during 60 min ischemia induced by periodic infusion of 50 to 100 microliter increments of air via the right internal carotid artery. The post-ischemic recovery of the CSER was followed an additional 60 min in 19 of these animals divided into 2 groups. Ten dogs were subjected to glass-wool filtration of their blood by extracorporeal shunting from femoral artery to femoral vein for one hr prior to infusing air. Nine dogs did not receive glass-wool filtration. Post-ischemic recovery of CSER amplitude, a quantifiable electrophysiologic index of neuronal function, was significantly greater in the filtered group than in the non-filtered group. 14C-antipyrine autoradiographic blood flow studies were performed in 3 dogs. One was studied at the end of a 60 min ischemic CSER suppression period and showed severe flow disruption by air embolism. Two dogs, one from each group, were studied at the conclusion of the 60 min recovery period. In the filtered animal, cortical blood flow exceeded the threshold for CSER maintenance while cortical flow rates in the unfiltered animal fell below this threshold. The enhanced postischemic neuronal recovery in the filtered group as indicated by the CSER in attributed to the preservation of injury zone nutrient blood flow that is supported by collateral circulation.

Afferent Pathways↗

Comparison of cotton wool swab and cytobrush for detection of chlamydial infection in women attending a genitourinary medicine clinic.

Endocervical specimens obtained by cytobrush and conventional cotton wool swabs from 90 women attending a genitourinary medicine clinic were compared for their efficiency in detecting chlamydial infection. Isolation of Chlamydia trachomatis and detection of the chlamydial lipopolysaccharide antigen were attempted on each specimen. Antigen was detected in 18% of cytobrush and 17% of swab specimens. The cytobrush proved less suitable than swabs for isolation because 8 cytobrush specimens (9%) were toxic to the McCoy cells. Toxicity was significantly associated with an infected endocervix (2P = 0.004). Cytobrush therefore appeared to have little advantage over the much cheaper alternative, the cotton wool swab.

Antigens, Bacterial↗

Retinal cotton-wool spots as the first sign of systemic sarcoidosis.

Isolated retinal cotton-spots have not yet been reported as the sign of ocular sarcoidosis. We describe three patients with retinal cotton-wool spots of initially unknown etiology in whom systemic sarcoidosis was diagnosed 6-16 months later. The diagnosis was confirmed by conjunctival biopsy. These observations indicate that sarcoidosis should be included in the differential diagnosis of retinal cotton-wool spots and patients with these lesions should be followed for the development of this disease.

Adult↗

No effect or no information? Comments on a nested case-control study of lung cancer among European rock and slag wool production workers.

The author considers the validity of a recent study of lung cancer among European rock and slag wool workers. The study failed to demonstrate an association between lung cancer and exposure to man-made vitreous fibers and also did not manage to demonstrate a relationship between lung cancer and asbestos exposure, an odd finding that casts doubt on its validity. This article deals with bias towards the null and other aspects of the reviewed study that may explain its failure to demonstrate an effect of asbestos, concluding that the study does not add to knowledge about a possible carcinogenic effect of rock and slag wool fibers, the apparent null results simply being non-informative because of the study's poor ability to detect existing associations.

Air Pollutants, Occupational↗

A mortality study of workers employed in a German rock wool factory.

As part of a coordinated European study including 13 plants, 2,096 workers employed in rock wool production, packing, and maintenance activities in a German factory were followed until 1979 for cause-specific mortality. There were no consistent differences between the observed and expected deaths in the study cohort on the basis of the experience of the general population. The separate analysis of persons engaged in rock wool production and packing and those engaged in maintenance yielded an excess of lung cancer deaths with borderline significance for occurrence among the maintenance workers and an elevation in standardized mortality ratio for stomach cancer related to time since first employment in the production group. Comparison with a factory-based reference cohort did not show any increased health risk for the exposed cohort. An extension of the follow-up for five years is planned in an attempt to clarify the present findings.

Construction Materials↗