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PubMed2003

Use of a major medical center clinical laboratory as a reference laboratory for a developing country: ordering patterns help set laboratory priorities.

BACKGROUND: The test menus for developed and developing countries may differ, depending on many factors, including the expected volume of testing, disease frequency and therapies available, clinical impact of the test, technical skill and equipment needed, cost, the patient population served, and whether alternative testing sites are available, and some of them may not be exactly known. We assessed test priorities in a developing country by making a broad range of tests available and then assessing which tests were actually used by the physicians in the country for the care of their patients. METHODS: The Barnes-Jewish Hospital laboratory and Washington University Medical Center provided patients in the developing country of Eritrea access to the same tests as patients in St. Louis for all analytes that are stable at 4 degrees C, the lowest temperature that could be used for shipping. RESULTS: The use of the St. Louis laboratories increased steadily from 1998 to 2001. More than one-half of the physicians in Eritrea used the reference laboratories, with requests for thyroid function and female fertility representing 48-71% of the test requests over the 4 years evaluated. The high degree of utilization for these test batteries was not predicted. Testing for thyroid function, female fertility, and lipid panels are now performed, or soon will be performed, in Eritrea based on the experience of the reference laboratory system. The reference laboratory system is continuing so that the test priorities of the country can be evaluated on an ongoing basis and specialized tests can be made available at a low cost. CONCLUSION: The experiences of a reference laboratory for a developing country can help to identify unanticipated priorities for medical testing within the country.

Clinical Laboratory Techniques↗
PubMed2003

Identification of AFLP markers linked to fertility restorer genes for tournefortii cytoplasmic male-sterility system in Brassica napus.

The tournefortii cytoplasmic male-sterility system is being used as a method of pollination control to develop hybrids in Brassica napus. Genetic analyses have indicated that two dominant genes, one major ( Rft1) and another minor ( Rft2), were required to achieve complete fertility restoration. Though the major gene ( Rft1) can cause complete fertility restoration on its own, its expression was significantly enhanced in the presence of the minor gene ( Rft2). In the absence of Rft1, Rft2 caused only partial fertility restoration. We used a pair of near-isogenic lines (NILs), differing for the presence/absence of Rf genes, to identify AFLP markers linked to fertility restorer genes. A total of 64 EcoRI/ MseI primer combinations were surveyed which produced 3,225 bands, of which 19 (0.006%) were polymorphic between parental NILs. Primer combinations which led to the identification of polymorphic bands present in fertile parental NILs were used for assaying a mapping population of 70 F(2) plants for determining the segregation pattern of markers. Initial screening resulted in the identification of five AFLP markers. The recombination analyses of these AFLP markers revealed that at least two (EACC/MCTT(105), EAAG/MCTC(80)) were present in the same linkage group along with the Rf loci. Marker EACC/MCTT(105) was separated from the major gene ( Rft1) by a distance of 18.1 cM, while it was 33.2 cM away from the minor fertility restorer gene ( Rft2). Another marker EAAG/MCTC(80) was also located adjacent to Rft1 at a distance of 18.1 cM, but on other side. Identification of flanking markers (EACC/MCTT(105), EAAG/MCTC(80)) for the major fertility restorer gene ( Rft1) provides a crucial component for marker-assisted selection and map-based cloning of the restorer genes, and can hence be used to construct elite restorer genotypes.

Brassica↗
PubMed1976

The fitness of human translocation carriers.

A simple method to estimate the mean fitness of human translocation carriers is described. The method is based on information about the inheritance pattern of the translocations. This kind of data can be obtained from the surveys of the chromosome constitutions of new-born babies. The following fitness estimates are obtained: D/D translocation carriers, 0-94; D/G translocation carriers, 0-83; and reciprocal translocation carriers, 0-67. The estimates are rather uncertain due to the scarcity of information, but they are consistent with the expected results. More accurate estimates will be obtained in the future when more data on translocation carriers are available and the cytological classification of the different translocations becomes more exact.

Biological Evolution↗
PubMed2000

[Capacitating action of GABA and progesterone in spermatozoa of human and guinea pig in vitro].

This study was designed to investigate whether GABA is involved in the capacitation effect and hyperactivated motility (HAM). Spematozoa from fertile men and retired guinea pigs were washed in modified BWW of 45% 90% Percoll gradient with 26 mg BSA/ml and in low Ca(2+)-MCM of 30%-55%-85% Percoll gradient (approximately 23 micromol/L Ca(2+)) respectively. The samples were preincubated for 2 h under 5% CO2 in air at 38.5 with or without GABA, progesterone (P(4)), GABA(A) receptor agonists or antagonists, and then exposed to 1 micromol/L (for human) or 5 micromol/L (for guinea pigs) calcium ionophore A 23187 for 15 min. The capacitation effect and HAM were assessed by using the chlortetracycline (CTC) staining method and phase-contrast microscopy. Motility was 80% 85% after all additions. The results showed that addition of GABA or P(4) at 5 micromol/L to the incubation medium resulted in a significant increase in the sum of B (characteristic of capacitated cell) and AR (acrosome recation) pattern (65.9% and 61.7% respectively), corresponding to capacitated spermatozoa in human, as compared to the control (37.3%). Likewise, the capacitating effect of GABA on spermatozoa in guinea pigs showed a concentration-dependent increase from 1 to 10 micromol/L(AR: 27.0 +/- 1.9% to 51.6 +/-2.8%). In addition, P(4) potentiated the capacitating effect of GABA when combined with GABA in the capacitation stage. The capacitating effect of GABA was mimicked by a GABAA receptor agonist muscimol. However, this effect was completely blocked by a GABA(A) receptor antagonist (-)-bicuculline and a GABA(A)/Cl(-) receptor antagonist picrotoxin. Furthermore, GABA markedly increased the HAM of P(4) on guinea pig spermatozoa, which was mediated obviously by the influx of extracellular Ca(2+) because the GABA-induced AR could be prevented by EGTA. These results indicate that GABA and P(4) are involved in the capacitation of spermatozoa in both human and guinea pigs through a GABA(A) receptor-mediated mechanism.

Acrosome Reaction↗
PubMed1988

Maintenance of foreign gene expression and independent chromosome fragments in adult transgenic rainbow trout and their offspring.

Transgenic rainbow trout (RT) were generated in earlier experiments by an in vivo modification of the chromosome-mediated gene transfer technique. This involved fertilizing albino RT eggs with gamma-irradiated brook (speckled) trout (ST) sperm and then heat shocking the eggs to induce second polar body retention. Striped pigment patterns were stable in six mosaic individuals between the ages of 7 and 14 months. We estimated the total ST DNA contribution in adult transgenic fish using DNA dot blot analyses. An average of 7% ST DNA was obtained in analyses of 11 transgenic individuals. Nine of 55 transgenic individuals examined expressed a ST-specific isozyme in adulthood; some were mosaic for foreign gene expression. Variable numbers of autonomous chromosome fragments were maintained in 10 adult transgenic individuals examined. Five transgenic RT were backcrossed to albino RT. Chromosome fragments were inherited by 29 of 31 backcross embryos examined. Additional active nucleolar organizer regions (NORs) were present on some fragments and may also have been integrated into some host chromosomes. This method may have numerous applications for basic and applied genetic research in fish.

Animals↗
PubMed2001

Three-dimensional modeling of hearing in Delphinus delphis.

Physical modeling is a fertile approach to investigating sound emission and reception (hearing) in marine mammals. A method for simulation of hearing was developed combining three-dimensional acoustic propagation and extrapolation techniques with a novel approach to modeling the acoustic parameters of mammalian tissues. Models of the forehead and lower jaw tissues of the common dolphin, Delphinus delphis, were created in order to simulate the biosonar emission and hearing processes. This paper outlines the methods used in the hearing simulations and offers observations concerning the mechanisms of acoustic reception in this dolphin based on model results. These results include: (1) The left and right mandibular fat bodies were found to channel sound incident from forward directions to the left and right tympanic bulla and to create sharp maxima against the lateral surfaces of each respective bulla; (2) The soft tissues of the lower jaw improved the forward directivity of the simulated receptivity patterns; (3) A focal property of the lower-jaw pan bones appeared to contribute to the creation of distinct forward receptivity peaks for each ear; (4) The reception patterns contained features that may correspond to lateral hearing pathways. A "fast" lens mechanism is proposed to explain the focal contribution of the pan bones in this dolphin. Similar techniques may be used to study hearing in other marine mammals.

Acoustics↗
PubMed2005

Effect of litter separation on 24-hour rhythmicity of plasma prolactin, follicle-stimulating hormone and luteinizing hormone levels in lactating rabbit does.

BACKGROUND: This work describes the effect of a 48-h litter separation on 24-h patterns of plasma prolactin, FSH and LH concentration in female lactating rabbits kept under a 16:8 light-dark photoperiod (lights on at 0800 h). METHODS: Groups of 6-7 female lactating rabbits maintained with their litters or separated from them for 48 h were killed by decapitation on day 11 post-partum, at 6 different time points throughout a 24-h cycle, starting at 0900 h. Plasma levels of prolactin, FSH and LH were measured by specific double antibody radio-immunoassays. RESULTS: Plasma level of prolactin in control and separated does changed in a similar way throughout the day, showing two maxima, at 0500-0900 h and at 1700-2100 h, respectively. Litter separation significantly augmented plasma FSH and LH and disrupted their 24-h rhythmicity. CONCLUSION: Since previous studies had shown that litter separation for short periods of time augmented sexual receptivity and fertility of the doe, the changes in FSH and LH reported may influence the massive release of gonadotropin releasing hormone, LH and FSH triggered by mating or artificial insemination in litter-separated mothers.

Journal Article↗
PubMed1983

The effect of pulsatile and continuous intravenous luteinizing hormone-releasing hormone administration on pituitary luteinizing hormone and follicle-stimulating hormone release in normal men.

In 14 healthy, potentially fertile men, pituitary gonadotropin responses were studied under standardized conditions. Luteinizing hormone-releasing hormone (LH-RH) was given as a continuous infusion of 1 microgram/minute for 4 hours or in a pulsatile fashion with 20 micrograms as an intravenous bolus at intervals of 20 minutes for 4 hours. Blood was collected continuously by means of an integrated sampling technique. The mean serum luteinizing hormone (LH) concentration showed an oscillating pattern around a plateau level reached within 45 minutes during continuous LH-RH administration. During pulsatile infusion, an identical pattern for the first 45 minutes was observed with, thereafter, a continuous increase from 105 minutes until the end of the infusion. The mean increase in the serum LH level during pulsatile administration was significantly higher (P = 0.00001) than the mean increase seen during continuous infusion. The follicle-stimulating hormone concentration revealed a gradual progressive increase after both methods of stimulation, without a significant difference in the mean increase between the two types of administration. This study demonstrates the existence of a self-priming effect of LH after pulsatile LH-RH administration in the man like that in the woman.

Adult↗
PubMed2004

No association of the A260G and A386G DAZL single nucleotide polymorphisms with male infertility in a Caucasian population.

BACKGROUND: The human DAZ gene family includes two autosomal genes, BOULE and DAZL, and a Y-chromosomal DAZ gene cluster. All are RNA-binding proteins and assumed to be master regulators of germline gene expression. We have investigated the impact of two DAZL polymorphisms, located at nucleotide positions 260 (SNP 260) and 386 (SNP 386), on the fertility of Caucasian men. These single nucleotide polymorphisms (SNPs) have been described previously to be associated with spermatogenic failure. METHODS: Blood samples were collected and genomic DNA was extracted from 165 normozoospermic men and 202 oligo- or azoospermic patients, of whom 28 displayed an AZFc deletion. The frequencies of A or G allelic variants in SNP 260 and 386 were analysed via TaqMan allelic discrimination assays. In both cases, the A to G transition leads to a threonine to alanine change. RESULTS: A total of 24.2% of the controls showed a heterozygous nucleotide variant (AG) for the SNP 260 and the remaining 75.8% were homozygous for A. In the AZFc-deleted group, this distribution was significantly different, with 39.3% for AG, 57.1% for AA and 3.6% for GG. However, the increased heterozygosity was not correlated with sperm counts and morphology. The patients without deletions displayed a similar allelic pattern to the controls (24.1% AG/75.9% AA). For SNP 386, only the AA nucleotide variant was found in all subjects studied and in no case was the previously described heterozygous AG variant found. CONCLUSION: In a selected Caucasian population, the DAZL SNP 386 is completely absent and SNP 260 is not associated with spermatogenic failure and therefore does not represent a molecular marker for genetic diagnosis of male infertility.

Alanine↗
PubMed1985

[Clinical use of vitamin A and E in gynecology].

Dysplasia is an alteration of organ and tissues cellular composition. It means quantitative and qualitative variations (differentiation) of the cells. More recently many Authors have investigated about epithelial neoplasia (breast, cervix, lung, respiratory tract, bladder, colon) and dysplastic epithelial processes and about their medical treatment and prevention of potentially transformative lesions. The view that certain vitamins such as vitamin A (in the form of retinol or its precursor beta-carotene) and vitamin E may protect against the risk of cancer has recently attracted much scientific attention. The potential use of retinoids (and alpha tocopherol) for chemoprevention has been demonstrated by numerous recent experiences. In gynaecology there are no evidence of therapeutical investigations about endometrial and ovarian dysplastic lesions, and then about protective role for consequent neoplasia development. Regarding cervical cancer there are only epidemiological retrospective data about dietary intake of vitamin A. The clinical trial is insufficient, perhaps for technical difficulties of administration, but we hope to obtain satisfactory results in the future. Benign breast disease, instead, so present in woman's life (in particular in fertile phase) is the most common clinical syndrome encompassing several distinct histopathological varieties. BBD is associated with an increase in breast cancer risk, particularly related to the presence of epithelial hyperplasia. In our study (collaborative study; University of L'Aquila--University of Rome) we have treated a group of women (double-blind trial) affected by breast dysplasia, with vitamin A and E association to evaluate the effectiveness of retinoids and alpha-tocopherol in reducing or resolving clinical palpable breast findings (with pain, tension, nodularity and instrumental patterns).(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Topical↗
PubMed1974

Population dynamics and age grading of mosquitos.

The dynamics of a vector population follows a characteristic pattern determined by the normal or modified environmental conditions of the biotope and by vector behaviour. The wide use of the precise method of age grading developed by Detinova and Polovodova has led to the conclusion that the dynamics of the various age groups may indicate behavioural, environmental, and epidemiological trends and changes and their significance. Age grading is useful for the qualitative assessment of vector density in general and of man-mosquito contact from the operational and epidemiological points of view, but only when sampling in time and space and the processing of the material collected have been correctly performed.

Age Factors↗
PubMed1992

[Marriage, marriage dissolution, and death 1976-1990: figures from life tables].

Life table data for the Netherlands are presented by marital status, age, and sex for four Dutch cohorts: never married, married, divorced, and widowed. "The numbers of survivors in each marital status are computed using sex, age and marital status specific rates...." Consideration is given to nonmarital fertility; average ages at first marriage, divorce, and remarriage; life expectancy at birth; and average age at death by sex and marital status. (SUMMARY IN ENG)

Age Distribution↗
PubMed1991

Uterine motility and cervical ripening in second trimester elective abortion by two different PGE analogues.

The clinical effects were studied of two different PGE analogues on the uterine motility and cervical ripening of eighty pregnant women asking for a second trimester elective abortion for fetal abnormalities. Forty women received vaginal suppositories each containing 1 milligram of 16, 16-dimethyl-trans-s2-PGE1 (Gemeprost) every 3 hours (5 mg max). Intramuscular injections of 500 micrograms of 16-phenoxy-w 17, 18, 19, 20 tetranor PGE2 methyl-sulphonylamide (Sulprostone) were administered every four hours (2000 mcg max.) to the remaining forty patients. Thirty-three Gemeprost treated patients (82.5%) and 34 Sulprostone treated patients (85%) experienced a complete abortion in the mean of 12.92 +/- 6.95 hours and 11.88 +/- 6.8 hours respectively. The histological and ultrastructural findings of cervical ripening were similar in both groups, while the tocographic patterns showed different characteristics. Side effects occurred in 16 Sulprostone (40%), but only in 9 (22.5%) Gemeprost treated patients, demonstrating that Gemeprost, although equally effective, is better tolerated.

Abortion, Induced↗
PubMed1982

Why is the number of pregnancies among teenagers decreasing?

The issue of pregnancy among adolescent women has received considerable attention from the media. Contrary to common belief, both the numbers and the rates of such pregnancies, even when data on abortion are included, have been declining. Patterns of contraception may account for some of the decrease; however, more study is required. In the past, unmarried teenagers who became pregnant either got married or put the baby up for adoption. Now they can either have an abortion or keep the baby. Solutions to the problems of pregnancy among teenagers must therefore be addressed to these altered social consequences rather than to misleading comments about "epidemics", with their suggestion of increased rates of pregnancy.

Abortion, Therapeutic↗
PubMed1995

[Diffusion of fertility control practices: moments, mechanisms, and determinants].

"In Brazil and in Bahia, distinct fertility control practices, performed through the so-called traditional techniques, have been socially diffused among the traditional elites and the more highly educated groups of the middle class since the end of the 19th century. These practices have produced a slow but systematic tendency to inter-generational decline in completed parity.... These facts support the hypothesis that the ethical changes involved in the acceptance and legitimacy of those practices were themselves a consequence of their increased prevalence and the consolidation of norms regarding the patterns of fertility control in these social classes.... Once anti-natality has been consolidated as a theoretical and practical reference system for the behaviour of the higher classes, the institutional apparatus of the society turned on the popular classes with lower levels of education and medical care, and diffused fertility control in these groups." (SUMMARY IN ENG)

Americas↗
PubMed1997

Characterization of calf follicular fluid and its ability to support cytoplasmic maturation of cow and calf oocytes.

The aims of the present study were to characterize the follicular fluid from prepubertal calf follicles of known size and quality and to study the ability of follicular fluid to support cytoplasmic maturation of calf and cow oocytes. Follicular fluid was obtained from 67 calf follicles classified according to size (S: small < 6 mm, M: medium 6-8 mm and L: large > 8 mm in diameter) and quality (HY: healthy, EA: early atretic and A: atretic). Quality was first determined by mitosis:pycnosis ratios in granulosa cell smears and confirmed by insulin-like growth factor binding protein (IGFBP) patterns. There was approximately 90% agreement between the two methods of follicle classification and on this basis the calf follicular fluid was pooled into nine groups. The accuracy of this pooling was confirmed by evaluation of oestradiol concentrations in the nine pools of follicular fluid using radio-immunoassay. Increases in follicle size were characterized by a decreased intensity of bands for IGFBP-2, IGFBP-5 and IGFBP-4, an increase in the proportion of healthy follicles and a decrease in the proportion of follicles in the early stages of atresia. This finding is in agreement with previously published results in cows. All classes of calf follicular fluid contained lower concentrations of oestradiol than previously reported for corresponding classes of cow follicular fluid. Cow oocytes were matured in M199 alone, or supplemented with 10% fetal calf serum (FCS), or 10% calf follicular fluid from one of three pools (LHY, LEA, LA), fertilized, and cultured for 8 days in synthetic oviduct fluid. Addition of FCS or calf follicular fluid to cow oocytes during in vitro maturation increased the yield of blastocysts on day 8 over the control (23%, 21/91), FCS (39%, 37/96, P < 0.05), LA (41% 21/52, P < 0.05), LEA (32%, 28/88), LHY (36%, 32/88), although not significantly in all cases. The rate of hatching of blastocysts was also improved: control (38%, 8/21), FCS (54%, 20/37), LA (62%, 13/21), LEA (75%, 21/28, P < 0.02), LHY (59% 19/32). In contrast, the addition of either FCS, calf follicular fluid or cow follicular fluid did not improve development of calf oocytes compared with the unsupplemented control. In conclusion, it is probable that serum and follicular fluid contain factors that stimulate the acquisition by oocytes, during maturation, of developmental competence and to which prepubertal oocytes are unable to respond. Specific receptors for these factors may develop only around puberty.

Animals↗
PubMed2004

Postpartum gonadotropin secretion in Holstein-Friesian dairy cows differing genetically in liveweight.

AIM: To determine whether there are differences in postpartum gonadotrophic activity between strains of Holstein-Friesian dairy cows genetically selected on mature liveweight that might explain differences between the strains in fertility, and the interval between calving and the resumption of ovarian follicular activity. METHODS: Mixed-age Holstein-Friesian cows fed generous allowances of ryegrass/white clover pasture, and genetically selected for heavy (H) or light (L) mature liveweight, were given 10 microg buserelin on Days 21, 28, 35 and 42 (Experiment 1a; n=8/group), or Days 7, 14, 21 and 28 (Experiment 1b; n=8/group) postpartum. The same dose of buserelin was also given to first-calved heifers from each strain (Experiment 1c; n=6/group) on Days 7, 14, 21, and 28 postpartum. Luteinising hormone (LH)concentrations were measured in serial blood samples that were taken for up to 240 min after administration of buserelin. In Experiment 2, serial blood samples were taken at 15-min intervals from H and L cows (n=7/group) over 8 h on Days 14, 21, 28 and 35 postpartum, to examine the endogenous secretion patterns of LH and follicle stimulating hormone (FSH). The time-course of the restoration of positive feedback between oestradiol and LH was examined by giving 1 mg oestradiol benzoate(ODB) 48 h after administration of 500 microg cloprostenol to mixed-age cows from each strain on Days 7 and 21 (n=8/group), or 14 and 28 (n=8/group) after calving (Experiment 3). Relationships between nutrition and the restoration of positive feedback were studied by giving 0.75 mg ODB/500 kg liveweight on Day 17 or 18 after calving to pure-bred Holstein (OSH) and New Zealand Friesian (NZF) cows that were fed either pasture (n=12 OSH, 12 NZF) or a total mixed ration (TMR; n=13 OSH,12 NZF) (Experiment 4). Plasma LH and FSH concentrations were measured in samples collected for 42 h (Experiment 3) or 48 h (Experiment 4) after treatment with ODB. Milk progesterone concentrations were measured 3x weekly to define the reproductive status of animals in each experiment. Conception rates were recorded for animals in all of the experiments. RESULTS: First-service conception rates were lower (p<0.05) in H than L cows (46% vs 59%). In Experiments 1b and 1c, LH response to buserelin increased between Days 7 and 28 postpartum (both p<0.001), but did not differ between strains (p=0.77 and p=0.19, respectively). In Experiment 1a, LH responses to buserelin did not change between Days 21 and 42 postpartum, but overall mean peak concentrations were significantly(p<0.001) greater in L than H cows. In Experiment 2, anoestrous H cows had higher mean (p=0.004) and episodic (p=0.001) concentrations of LH than did L cows, but in cows that had active corpora lutea there were no such differences. There were no differences in FSH concentrations between strains. LH secretion in response to exogenous oestradiol (Experiment 3) increased between Days 7 and 28 postpartum (p<0.001), but there were no differences between strains. Responses were also similar in OSH and NZF cows on Day 17 or 18 postpartum, although there was a significant effect of ration upon the proportion of cows that exhibited an LH surge (20/24 cows on grass vs 12/25 on a TMR; p=0.005). CONCLUSION: These results confirm that H cows have poorer first-service conception rates than L cows, but do not support an hypothesis that there are major differences between these strains of Holstein-Friesian dairy cows in the rate of restoration in the hypothalamo-pituitary axis. However, in anoestrous cows, differences between strains in the endogenous release of LH maybe related to an earlier onset of oestrous cycles in H animals.

Journal Article↗