Testis development and gonadotropin secretion in broiler breeder males.

One-day-old chicks were used to evaluate testis development and concentrations of luteinizing hormone, follicle-stimulating hormone (FSH), and testosterone during ontogenesis. Males on a conventional breeder program (control) were reared on a 15L:9D photoperiod and ad libitum food and water intake for 2 wk. On the third week, males were placed on a restricted diet and the photoperiod was reduced to 8L:16D. Males on a pedigree breeder program (broilerized) were reared on a 23L:1D photoperiod and unrestricted food and water intake for 6 wk. At 7 wk, males were placed on a restricted diet and the photoperiod was reduced to 8L:16D. On wk 18, both treatment groups were photostimulated (16L:8D) until the end of the experiment (50 wk). After photostimulation, there was an exponential increase in testis weight (TW), FSH, and testosterone concentrations. At 28 wk, TW from broilerized males were significantly heavier than those from control birds, and concentrations of luteinizing hormone, FSH, and testosterone were maximal at that time. After 28 wk, there was a significant decrease in FSH and testosterone concentrations that were associated with reduced TW. No correlation was observed between BW and TW. However, TW was highly correlated with FSH concentrations and daily sperm production. Our data suggest that management and photoperiod had a profound effect on testicular function that was associated with FSH concentrations in male broiler breeders.

Active immunization against ghrelin decreases weight gain and alters plasma concentrations of growth hormone in growing pigs.

Ghrelin has been implicated in the control of food intake and in the long-term regulation of body weight. We theorize that preventing the ability of ghrelin to interact with its receptors, would eventually lead to decreased appetite and thereby decrease body weight gain. To test our hypothesis, pigs were actively immunized against ghrelin. Ghrelin((1-10)) was conjugated to BSA and emulsified in Freund's incomplete adjuvant and diethylaminoethyl-dextran. Primary immunization was given at 19 weeks of age (WOA), with booster immunizations given 20 and 40 days after primary immunization. Body weight (BW) and plasma samples were collected weekly beginning at 19 WOA, and feed intake was measured daily. Fourteen days after primary immunization, the percentage of bound (125)I-ghrelin in plasma from immunized pigs was increased compared with control animals (P<0.001). Voluntary feed intake was decreased more than 15% in animals that were actively immunized against ghrelin compared with controls. By the end of the experiment, immunized pigs weighed 10% less than control animals (P<0.1). Concentrations of GH were increased (P<0.05) in immunized pigs. Apoptosis was not observed in post-mortem samples obtained from the fundic region of the stomach. Our observations suggest that immunization against ghrelin induces mild anorexia. This procedure could potentially be used as a treatment to control caloric intake and obesity.

An inexpensive, simple protocol for DNA isolation from blood for high-throughput genotyping by polymerase chain reaction or restriction endonuclease digestion.

We describe simple, inexpensive, and reliable methods for isolating DNA from avian blood, semen, or feather pulp. The procedures are readily applicable to high-throughput 96-well plate isolation for genotype analysis of chicken DNA based on restriction endonuclease digestion or PCR. Isolation cost is primarily the cost of a deep-well assay block and a few pipet tips; current price is less than 0.10 dollar per sample, providing a significant cost advantage over commercial kits. The procedure employs inexpensive, nonhazardous reagents and yields intact, double-stranded DNA from as little as 2 to 10 microL of avian blood, suitable for RFLP analysis or hundreds of PCR amplifications. We compared our method to published procedures for alkaline extraction from feather pulp and found our method to be more reliable with the advantage of isolating intact DNA sequences that can be easily quantified. With minor modifications, the method can isolate DNA for PCR genotyping from mammalian whole blood.

An expressed sequence tag analysis of the chicken reproductive tract transcriptome.

Analysis of the chicken reproductive tract transcriptome is important in comparative biology for analysis of reproductive tract development and evolution. In addition, molecular analysis of the reproductive tract is important for identification of genes affecting fertility in the poultry industry. We sampled the chicken reproductive tract (ovary, oviduct, and testis) transcriptome, generating 5,328 expressed sequence tags that assembled into 4,518 contigs. We identified 475 contigs with no match in the current expressed sequence tag databases or in GenBank. The novel contigs included 31 with no match to the current assembly of the chicken genome, 119 representing spliced transcripts, and 309 that were unspliced. More detailed molecular characterization of the 428 novel contigs present in the assembly will be important to gene discovery and annotation of the chicken and other vertebrate genomes.

Adult testicular enlargement induced by neonatal hypothyroidism is accompanied by increased Sertoli and germ cell numbers.

Our previous studies have shown that transient neonatal hypothyroidism, induced by treatment with the reversible goitrogen 6-propyl-2-thiouracil (PTU), increases testicular size and daily sperm production in the adult rat by up to 82% and 136%, respectively. The objective of the present study was to examine morphological and functional changes in adult seminiferous tubules associated with PTU-induced increases in testicular size and sperm production. Sprague-Dawley rats were treated with PTU from birth to day 25 or left untreated; for morphometry, all testes were fixed by vascular perfusion at 90 days of age. Although testicular weight was increased 62% in treated rats, gross pathological changes were not evident in these organs, and spermatogenesis appeared morphologically normal. The percent area of testis occupied by seminiferous tubules was equal in control and treated testes, but mean seminiferous tubule diameter and length were increased in the PTU-treated testis. The adult number of Sertoli cells in treated testes was increased by 157%, and the numbers of leptotene spermatocytes and round spermatids were increased 84% and 93%, respectively. These results demonstrate that increases in Sertoli cell numbers result in increased sperm production and support the idea that Sertoli cells are the major regulators of the magnitude of sperm production. Although the round spermatid to Sertoli cell ratio was reduced by nearly 30%, the number of round spermatids per g testis was increased by 14%. This increased efficiency of sperm production was accomplished by an increased density of Sertoli cells along the basement membrane and an increased height of the seminiferous epithelium. Despite the large increase in Sertoli cell numbers in treated rats, Northern blot analysis using Sertoli cell-specific cDNA probes for transferrin and androgen-binding protein indicated that relative steady state levels of mRNAs per Sertoli cell for these two secretory proteins were similar in control and treated rats at 90 days of age.

Leydig cells increase their numbers but decline in steroidogenic function in the adult rat after neonatal hypothyroidism.

Administration of the goitrogen, 6-propyl-2-thiouracil (PTU), to suckling rat pups from birth through day 24 postpartum as a 0.1% solution in the mother's drinking water increases adult testis size and sperm production by about 80% and 140%, respectively, without affecting peripheral testosterone levels. The objectives of this study were to determine whether adult Leydig cell numbers were altered in PTU-treated rats and whether the steroidogenic function of these cells was normal. The number of Leydig cells per testis at 180 days increased by 69% in PTU-treated compared to control rats, whereas the average Leydig cell volume declined by about 20%. Steroidogenic function assessed in isolated adult Leydig cells decreased after neonatal PTU treatment. LH-stimulated testosterone production was reduced by 55% in Leydig cells from treated rats, commensurate with a 50% decline in the number of hCG-binding sites in these cells. The difference in steroidogenic potential was even more striking after incubations with saturating concentrations of steroid substrate, 22(R)-hydroxycholesterol; Leydig cells from treated males produced 73% less testosterone than controls. Therefore, this decrease in testosterone production may be partially due to a reduction in the numbers of LH receptors, but also reflects the impaired steroidogenic potential of these cells. These results clearly show that the dramatic increase in adult Leydig cell number after neonatal PTU treatment is counterbalanced by a permanent decline in Leydig cell steroidogenic function, producing no net change in peripheral testosterone levels.

Developmental hormonal profiles accompanying the neonatal hypothyroidism-induced increase in adult testicular size and sperm production in the rat.

Neonatal treatment with the reversible goitrogen 6-N-propyl-2-thiouracil (PTU) results in a near doubling of testicular size and a 25% increase in the efficiency of spermatogenesis, without affecting circulating testosterone (T) levels in adult rats. The objectives of the present study were to examine the effects of neonatal PTU treatment on the pattern of testicular growth and circulating levels of anterior pituitary (FSH, LH, PRL, GH, and TSH), gonadal [immunoreactive inhibin-alpha (irI alpha) and T], and thyroid (T3 and T4) hormones over the first 100 days of life. Treatment of rats with PTU from birth to 24 days of age significantly reduced testicular weights between 10 and 60 days of age. However, the duration of testicular growth was extended in treated males, resulting in a 68% increase at 100 days of age. Serum gonadotropin levels in treated males were reduced throughout the experimental period, typically remaining between 50-70% of control levels. The characteristic robust prepubertal FSH peak was absent in PTU-treated males. Initially high until 20 days of age, irI alpha levels characteristically declined to adult levels (200-300 pg/ml) in control males. In treated males, irI alpha levels were reduced during the period of hypothyroidism, increased between 30 and 60 days, and then declined, but remained significantly higher (1.7- to 2-fold greater) than those observed in control males. Serum T levels were similar in treated and control males. Control males demonstrated increased T levels beginning at 45 days of age, earlier than observed in treated males; however, similar peak T levels were observed in all males. PTU treatment significantly suppressed serum GH and PRL and led to a 14-fold increase in circulating TSH during the period of treatment. However, unlike the gonadotropins, these hormones returned to control levels after PTU treatment, suggesting that the reduced levels of FSH and LH observed are not due to a generalized reduction in pituitary function. Serum T4 and T3 levels returned to control levels within 15 days after the removal of PTU. These results demonstrate that the neonatal PTU treatment-induced increases in adult testicular size and sperm production were not due to increased levels of FSH at any point in development. On the contrary, the observed increases occur in spite of chronically reduced FSH levels.

Neonatal hypothyroidism permanently alters follicle-stimulating hormone and luteinizing hormone production in the male rat.

Transient neonatal hypothyroidism, induced with the goitrogen 6-n-propyl-2-thiouracil (PTU), results in dramatic increases in both testis size and sperm production in the adult rat. The observed increases in testis size and function occur in the presence of normal circulating testosterone levels. However, circulating gonadotropin levels are chronically reduced by 30-50% at all times in treated males. To better understand the permanent reduction in serum gonadotropin levels following transient neonatal hypothyroidism, we conducted a series of experiments to evaluate pituitary and hypothalamic function in the adult male PTU-treated rat. PTU treatment led to a significant reduction in GnRH-stimulated LH production. Castration resulted in 3.9- to 8.5-fold increases in circulating gonadotropin levels in both treated and control males; however, the absolute increases were significantly reduced in treated males. In contrast to circulating levels, pituitary gonadotropin contents did not increase in treated males after castration. PTU treatment did not lead to a reduction in the density of either luteotropes or folliculotropes, and both cell types increased in size and density after castration. The relative concentrations of both gonadotropin beta-subunit messenger RNAs increased more slowly in treated males than in controls after castration. Thus, although treated rats have the intrinsic ability to produce normal circulating levels of LH and FSH, gonadal feedback and an overall reduction in gonadotrope synthetic ability combine to produce the chronically reduced circulating levels of these hormones.

Infusion of a stable prostacyclin analogue, iloprost, to patients with peripheral vascular disease: lack of antiplatelet effect but risk of thromboembolism.

PURPOSE: Prostacyclin, a potent inhibitor of platelet function and vasodilator, has been used to treat peripheral vascular disease. The aim of this study was to monitor the thrombotic status of patients treated by infusion of a stable prostacyclin analogue, iloprost. PATIENTS AND METHODS: Thirteen patients with peripheral vascular disease underwent iloprost infusion for 3 days (8 hours each day) in a dose ranging from 0.5 to 2 ng/kg/minute. Variable parameters of thrombosis such as platelet reactivity (shear-induced hemostatic plug formation and thrombus formation on a collagen fiber), coagulation, and spontaneous thrombolysis (dislodgment of hemostatic plugs) were measured from non-anticoagulated blood samples by hemostatometry immediately before and 1 hour after the infusion and on the last day, 4 hours after initiation of the infusion. RESULTS: Analysis of data from all patients 1 hour after the infusion showed no changes in platelet reactivity and spontaneous thrombolysis, but coagulation was significantly enhanced. In four patients, significant platelet hyperreactivity was observed after the infusion. Four of the five patients tested while undergoing iloprost infusion showed an enhanced thrombotic reaction and markedly enhanced coagulation. Iloprost employed in vitro in a concentration that corresponds to the therapeutic peak blood level caused no inhibition of platelet function but significantly enhanced coagulation. The threshold in vitro iloprost concentration at which anti-platelet effect and increased spontaneous thrombolysis were observed was twice that of the therapeutic blood level. CONCLUSIONS: These findings challenge the view that antagonism of platelet function is an important factor of iloprost therapy. Furthermore, platelet hyperreactivity in some patients and markedly enhanced coagulation during and after infusion of iloprost in general, represent a risk of thromboembolism, especially as patients are already in a prethrombotic condition.

A cholinergic antagonist, mecamylamine, blocks light-induced fos immunoreactivity in specific regions of the hamster suprachiasmatic nucleus.

Recent studies have shown that light-induced phase shifts of the circadian rhythm of locomotor activity are associated with c-fos expression in the suprachiasmatic nucleus (SCN) of rodents. In order to explore further the importance of c-fos expression for the phase-shifting effects of light, we examined the effects of mecamylamine on light-induced Fos-like immunoreactivity (Fos-lir) in the SCN. Mecamylamine was chosen because it is a cholinergic antagonist that blocks the phase-shifting effects of light on the circadian activity rhythm in the golden hamster. Golden hamsters were entrained for at least 14 days to a 14 h light: 10 h dark photoperiod. Animals were then placed in constant darkness (DD) and during exposure to DD were subjected to one or more of the following treatments: (1) vehicle alone; (2) mecamylamine alone (450 micrograms, i.c.v.) at circadian time (CT) 19; (3) vehicle plus a light pulse at CT 19; (4) mecamylamine 10 min prior to the light pulse at CT 19. Mecamylamine blocked the phase-shifting effects of light on the circadian rhythm of locomotor activity when compared to vehicle-treated animals (P < 0.001). A light pulse at CT 19 induced Fos-lir in the SCN within 1 h, whereas treatment with vehicle or mecamylamine had no noticeable effect on Fos-lir in the SCN. Mecamylamine pretreatment dramatically reduced light-induced Fos-lir in the SCN by 75%. The most striking observation was the clear inhibition of Fos-lir by mecamylamine in the dorsomedial region of the SCN while there was little inhibition of Fos-lir in the most ventral portions of the SCN.(ABSTRACT TRUNCATED AT 250 WORDS)

Estrogen receptors are present in the epididymis of the rooster.

Our recent discovery that testicular germ cells and epididymal sperm contain active P450 aromatase suggests that the reproductive tract may be a target for estrogen. Therefore, the objective of this study was to determine if estrogen receptors (ER) are present in the avian epididymis using immunocytochemistry, northern blot analysis, and in situ hybridization. Immunoperoxidase staining for ER was found principally in nuclei of nonciliated epithelial cells of proximal and distal efferent ductules and the epididymis duct. The ciliated cells also appeared to be slightly positive in the efferent ductules. Week immunostaining was also observed in the connective tissue of the epididymis duct. Immunostaining was more intense in epithelial cells of the efferent ductules than in epithelial cells of the epididymal duct of connective tissue cells. Strong specific hybridization signals for ER mRNA corresponded to the same areas exhibiting immunocytochemical localization. The presence of ER mRNA in the epididymis was confirmed by northern blot analysis, which showed a single band corresponding to approximately 7.8 kb, similar to that found in chicken oviduct. Based on these data, we suggest that the efferent ducts of the rooster are a primary target for estrogen and that estrogen may have a role in the regulation of avian epididymal function.

Onset of spermatozoal degeneration in low-fertility Delaware roosters and test for autoimmune basis.

The objectives of this study were 1) to determine the onset of a heritable reproductive disorder in the rooster that is characterized by extensive spermatozoal degeneration within the ductus deferens, and 2) to determine if autoimmunity was associated with spermatozoal degeneration. Seventy-five percent of the affected roosters did not ejaculate large percentages of degenerate spermatozoa at 20 wk of age, approximately the age of sexual maturity. Rather, seminal quality gradually declined over the next 6 wk, as both ejaculate volume and number of spermatozoa ejaculated increased. The evaluation of testicular and excurrent duct tissues via immunofluorescence failed to reveal either IgY or IgA associated with spermatozoa. While histological examination revealed greater lymphocyte numbers (P less than .05) in the proximal ductus deferens, these cells were not associated with spermatozoa nor spermatozoal clumping. While spermatozoal degeneration tends to be latent at the onset of semen production, it does not appear to be due to spermatozoal autoimmunity.

Calcium influx into red blood cells: the effect of sera from patients with systemic sclerosis.

The rheology of red blood cells in patients with systemic sclerosis is abnormal. To investigate this further we have examined the effect of sera from patients with systemic sclerosis on the handling of calcium ions by the erythrocyte membrane. Normal erythrocytes were filled with a photoprotein (aequorin) which emits light on contact with calcium. These photoprotein loaded normal erythrocytes were then incubated overnight with serum from: normal subjects (n = 20), from patients with systemic sclerosis (n = 27) or from patients with primary Raynaud's disease (n = 10). There was no significant difference in basal calcium leakage, as measured by the amount of light produced following the addition of triton X-100. Induced calcium influx, as measured by the amount of light produced following the addition of ionophore A23817, was significantly greater in the photoprotein loaded erythrocytes incubated overnight with serum from patients with systemic sclerosis compared to those incubated with serum from normal subjects (p less than 0.02) or patients with primary Raynaud's disease (p less than 0.01). This modulation of Ca2+ handling in erythrocytes by a serum factor from patients with systemic sclerosis could account for the alterations in erythrocyte function, such as red cell deformability, observed in systemic sclerosis.

The anti-platelet effect of nifedipine in patients with systemic sclerosis.

The enhanced platelet reactivity and impaired thrombolysis in patients with systemic sclerosis may contribute to the microvascular insufficiency seen in this disease. Anti-platelet therapy has therefore been suggested for the treatment of Raynaud's phenomenon secondary to systemic sclerosis. Using a novel technique, the Haemostatometer, haemostasis (shear-induced) and thrombolysis (dislodgement of the haemostatic plug) were assessed initially in 15 patients with systemic sclerosis before and 90 minutes after a single oral dose of nifedipine (10 mg), and then at 4-week intervals for 16 weeks in 10 patients on long-term nifedipine (10-20 mg tid). Ninety minutes after a single oral dose of nifedipine in 15 patients with systemic sclerosis, haemostasis was significantly prolonged from 140 +/- 12 sec to 178 +/- 21 sec (mean +/- SE, p less than 0.005). The time until spontaneous thrombolysis occurred was significantly shortened following nifedipine, from an abnormal time of 57.8 +/- 2.8 min to a more normal value of 34.0 +/- 5.1 min (mean +/- SE, p less than 0.01). This improvement in haemostasis and thrombolysis was maintained through 16 weeks on longterm nifedipine treatment. These findings suggest that nifedipine may reduce the risk of developing multiple thrombo-emboli in patients with systemic sclerosis.

Effects of 6-N-propyl-2-thiouracil on growth, hormonal profiles, carcass and reproductive traits of boars.

Neonatal 6-N-propyl-2-thiouracil (PTU)-induced hypothyroidism reduces body weight but increases testicular size in adult male rodents. The objective of this study was to determine the effect of prepubertal PTU treatment on boars. For Experiment I, boars (n = 28) were randomly allotted to eight pens. Each pen received one of four PTU doses (0, 0.01, 0.03 and 0.1% in a basal diet) between 28 and 56 days of age (DOA). Due to a lack of difference among three PTU treatments, PTU-treated boars were pooled. Boars treated with PTU had lower (P < 0.05) ADG during treatment, lighter (P < 0.05) BW after 56 DOA and less (P < 0.05) developed epididymides at 154 DOA. For Experiment II, boars (n = 19) were randomly allotted to six pens. Each pen received one of three PTU treatments orally as: control (carrier), PTU-I (0.002% BW of PTU daily between 7 and 70 DOA), or PTU-II (0.002% BW of PTU daily between 28 and 91 DOA). During treatment, PTU-treated boars had lower (P < 0.05) serum T4 levels, rectal temperature, feed intake and ADG. Boars treated with PTU had lower (P < 0.05) BW between 63 and 154 DOA but higher (P < 0.05) gain/feed between 105 and 133 DOA. Boars treated with PTU had less (P < 0.05) developed epididymides and sperm count per gram testis at 238 DOA. These results suggest that prepubertal PTU-induced hypothyroidism had significant effects on growth, hormonal profiles, and reproductive traits of boars; however, it does not appear to be an effective method for increasing testis size and sperm production of commercial boars.

Treatment of severe Raynaud's syndrome by injection of autologous blood pretreated by heating, ozonation and exposure to ultraviolet light (H-O-U) therapy.

OBJECTIVE: To determine the effect of re-injection of small samples of autologous blood, pretreated with heat, ozone and ultraviolet light (H-O-U therapy) in patients with severe Raynaud's syndrome. EXPERIMENTAL DESIGN: Open trial in 4 patients. SETTING: Temperature/humidity controlled vascular laboratory. PATIENTS: Severe Raynaud's syndrome of more than 5 years duration and defined as more than 5 attacks daily or 10 attacks in one week, at least half of which were painful and lasting for more than 30 minutes. Three patients were refractory to infusions of Iloprost. INTERVENTIONS: Patients were treated daily or on alternate days for a two to three weeks period by re-injection of citrated autologous blood pre-treated with heat, ozone and ultraviolet light (H-O-U therapy). MEASURES: Clinical observations; mean equilibrated hand temperature (infrared thermography); distributive and microcirculatory blood-flow (venous occlusion strain-gauge plethysmography, infrared photoplethysmography, laser Doppler flowmetry) iontophoresis of acetylcholine and sodium nitroprusside; estimations: serum levels of 6-keto-PGF1alpha and serum levels of anti-hsp65 antibody. RESULTS: Reduction or abolition of Raynaud's attacks for at least three months after treatment. Mean equilibrated hand temperature increased but did not normalise. Blood flow parameters improved but did not reach statistical significance. Iontophoresis of acetylcholine showed an increase in laser Doppler flowmetry which was statistically significant. Serum levels of 6-keto-PGF1alpha, fell significantly in three patients. Serum levels of anti-hsp65 antibody fell in the one patient which was followed sequentially. CONCLUSIONS: H-O-U therapy may prove useful in patients with severe Raynaud's syndrome.

Hemostatic abnormalities in Sneddon's syndrome.

Sneddon's syndrome is a rare condition comprising widespread livedo retucularis and multiple episodes of transient cerebral ischemia. Treatment to date has been empirical. The hemostatic/thrombotic status of 4 patients with Sneddon's syndrome was studied by a unique technique, hemostatometry, which measures primary hemostasis (shear-induced platelet plug formation), the overall coagulation, and thrombolysis (dislodgment of the hemostatic plugs) from nonanticoagulated blood. In all 4 patients, platelet reactivity, which shows itself in the initial phase of the hemostatic reaction, was enhanced. The overall hemostasis, in which the generation of thrombin by activated platelets plays the decisive role, was enhanced in 3 patients. Three of the 4 patients had hypercoagulation, and in 3, spontaneous thrombolysis was inhibited. Treatment was commenced with aspirin and nifedipine, and patients were monitored both clinically and by serial hemostatometry over two years. One patient had one further transient ischemic episode; the other 3 remained asymptomatic. Thus, the observed clinical improvement correlated with improvement of the hemostatic profile.

Altered in vitro hemostasis, clotting, and thrombolysis after oral nifedipine in normal volunteers.

In this study the authors used the Haemostatometer, a new instrument to monitor the pattern of hemostatic plug formation that occurs in holes in polyethylene tubing through which nonanticoagulated blood is flowing under standard conditions. The pattern and speed of blood coagulation, subsequent to hemostasis, was also monitored. Simultaneously, the time until expulsion of hemostatic plugs formed was measured and considered as spontaneous thrombolysis time (STT). In 10 healthy volunteers, blood samples were drawn and tested before and ninety minutes after administration of 10 mg oral nifedipine. After nifedipine, the initial phase of primary hemostasis was delayed (p less than 0.05), the clotting time lengthened (p less than 0.01), and the STT shortened (p less than 0.01). The authors conclude that the effect of nifedipine on hemostasis and thrombolysis could contribute to its therapeutic efficacy.

CL115,347, an analogue of prostaglandin E2. Peripheral circulatory effects of single ascending doses administered transdermally in normal subjects and in patients with Raynaud's phenomenon.

The beneficial effects of intra-arterial or intravenous infusion of the prostanoid products of anachridonic acid, PGE and prostacyclin (PGI2) are well documented. More recently an analogue of PGE2, (CL 115,347, American Cynamid Co.) has become available. This substance is absorbed transdermally from a patch placed on the skin. In a placebo-controlled trial the vasodilatory effect of single incremental dosage, 500 mcg, 1000 mcg and 1500 mcg, was measured in a temperature-/humidity controlled laboratory in normal subjects and in patients with primary and secondary Raynaud's phenomenon. The optimal dosage proved to be 1000 mcg; the effect may last for 84 hours; higher dosage may be associated with a "steal" phenomenon.

Analysis of poultry fertility data. 2. Comparison of long- and short-term fertility trials.

The purpose of the present study was to determine the effect of egg collection interval on interpretation of fertility trial results. Low-fertility (LF) and randombred (RB) Delaware hens were inseminated with spermatozoa from LF or RB Delaware roosters. Three replicate trials were performed for each cross. Eggs were collected for 21 days following insemination. Fertility was analyzed with a log odds model following logit transformation. Separate analyses were performed on data collected throughout the first 7 days and the entire egg collection interval. Duration of fertility over the 21-day interval was analyzed by iterative least squares. Whereas the 21-day egg collection interval afforded detection of differences between males and between females, no differences were observed following analysis of data from the 7-day interval. Differences in reproductive efficiency also were detected when data were analyzed by iterative least squares. Thus, the value of a fertility trial is, in part, determined by the length of the egg collection interval.