Elevation of plasma tyrosine after a single oral dose of L-tyrosine.

Plasma tyrosine concentrations in twelve normal, fasting human subjects were significantly elevated 2–8 hours after they ingested 100 mg/kg or 150 mg/kg tyrosine. Mean plasma tyrosine levels were maximal after 2 hours, rising from $\text{69 ± 3.9}\text{to}\text{154 ± 9.5}\text{nmols/ml}\text{(}\text{X}\text{±}\text{SEM}\text{)}$ after the 100 mg/kg dose and to 203 ± 31.5 nmols/ml after the 150 mg/kg dose (p ≤ 0.001 for both doses). The mean tyrosine ratio (defined as the ratio of plasma tyrosine concentration to the sum of the concentrations of six other neutral amino acids that compete for the same blood-brain barrier uptake system) increased from $\text{0.10 ± 0.02}\text{to}\text{0.28 ± 0.04 (}\text{X}\text{±}\text{SEM}\text{)}$ 2 hours after the 100 mg/kg dose (p ≤ 0.001) and to 0.35 ± 0.05 2 hours after the 150 mg/kg dose (p ≤ 0.005). No side effects of orally-administered L-tyrosine were noted.     

Pregnancy detection in tapirs by direct urinary estrone sulfate analysis

Urinary levels of estrone sulfate (ES), indexed by creatinine (CR), were evaluated by a direct radioimmunoassay of four Malayan (Tapirus indicus) and one Brazillian (Tapirus terrestris) tapir pregnancies. Levels rose above baseline ES values of 20 ± 1.2 ng/mg CR (n = 200) in the Brazilian animal and 25 ± 1.3 ng/mg CR (n = 105) in the Malayan animals at approximately 7 months prior to parturition and continued to rise in both species until just before parturition. Quantitatively, levels rose 10-fold higher in the Brazillian animal than in the Malayan animals through approximately 1 month prior to parturition in both species. These findings indicate that routine urinary monitoring provides an accurate means for detecting pregnancy in tapirs and suggests differences in estrogen excretion patterns between tapir species and with other perissodactyls.     

Urinary steroid and gonadotropin excretion across the reproductive cycle in female Wied's black tufted-ear marmosets (Callithrix kuhli)

Details of the endocrinology of reproduction in the genus Callithrix are known only for the common marmoset, C. jacchus. This paper presents the patterns of urinary pregnandiol-3-glucuronide (PdG), urinary estrone conjugates (E₁C), and gonadotropin excretion throughout the reproductive cycle of Wied's black tufted-ear marmoset (C. kuhli) as determined via steroid conjugate enzyme immunoassays (EIA) and gonadotropin radioimmunoassays (RIA). Postpartum ovulation occurred at 13.6 ± 1.2 days after parturition (n = 12) and was characterized by low PdG and E₁C concentrations accompanied by a spike in luteinizing hormone (LH)/chorionic gonadotropin (CG) concentration. After conception, PdG concentrations increased dramatically until they dropped to periovulatory concentrations in the third trimester of pregnancy. Mean PdG concentrations in the first and second trimesters (33.7 ± 8.4 and 39.0 ± 10.9 μg/mg creatinine, respectively) were three times that of third trimester concentrations (11.7 ± 1.4 μg/mg Cr; n = 8). Urinary concentrations of E₁C rose more gradually during pregnancy and remained higher prepartum than urinary concentrations of PdG. Urinary gonadotropin concentrations also increased after conception (first trimester concentrations = 24.5 ± 4.5 ng/mg Cr) and continued to increase in the second trimester (51.4 ± 7.6 ng/mg Cr), until they finally decreased in the third trimester (mean = 7.9 ± 1.4 ng/mg Cr; n = 8). The interbirth interval was 156.3 ± 2.9 days (n = 6), with a gestation of 143.1 ± 1.6 days (n = 8). Nonconceptive cycle length was 24.9 ± 0.6 days (n = 4). The results of this study suggest strong similarities in reproductive parameters in the genus Callithrix. © 1996 Wiley-Liss, Inc.     

Kinetics of testosterone metabolism in normal postmenopausal women and women with breast cancer

The constant infusion and single injection techniques were utilized to study the kinetics of ³H-testosterone (T) metabolism in postmenopausal women with and without breast cancer. The metabolic clearance rates (mean ± SEM) for normal postmenopausal women were $\text{578 ± 82}\text{and}\text{644 ±128}\text{1}\text{24}\text{h}$ as obtained by the constant infusion and single injection techniques, respectively. The corresponding results for the women with breast cancer (patients) are $\text{644 ± 25}\text{and}\text{617 ± 106}\text{1}\text{24}\text{h}$. The single injection technique yielded values for rate constants (units) and volumes of distribution (1); k₁ = 37.5 ± 1.6 for the normals and 34.5 ±1.9 for the patients. K₂ = 76.6 ± 5.1 for the normals and 71.1 ± 1.6 for the patients, V₁ = 7.9 ± 2.2 for the normals and 8.7 ± 1.4 for the patients and V₂ = 7.0 ± 1.5 for the normals and 6.4 ± 1.2 for the patients. The constant infusion technique yielded values for the conversion ratios for the transformation of T to several products; 4-androstene-3,17-dione/T of 0.02 ± 0.003 for normals and 0.03± 0.002 for patients, 5α-dihydrotestosterone/T of 0.02 ± 0.002 for normals and 0.03 ± 0.002 for patients, estrone/T of 0.04 ± 0.01 for normals and 0.04 ± 0.01 for patients, estradiol-17β/T of 0.02 ± 0.005 for normals and 0.03 ± 0.005 for patients and estrone sulfate/T of O.16 ± 0.02 for normals and 0.24 ± 0.06 for patients. The T plasma concentrations and production rates were similar for the two groups of subjects. Hence there were no significant differences between the normals and the patients for all the kinetic parameters. It was determined that all the estradiol being produced in postmenopausal women could be coming from circulating T.     

Direct estimation of urine estrone conjugate for a rapid pregnancy diagnosis in sows

A rapid and direct radioimmunoassay of urine estrone conjugate (E₁C) was developed. Urine samples were taken from Large White sows by inserting sponges in the vagina which were expelled during micturition. Analysis of samples collected daily from 13 sows between 2 and 40 days after service showed that high urine E₁C levels occurred between day 20 and 30 after insemination in pregnant sows; maximum concentrations were observed on day 25 (79.4 ± 11.7 ng/ml, $\text{X}\text{± s.e.m.}$). In this period E₁C levels never exceeded 1 ng/ml in nonpregnant sows. A single urine sample was then taken from 84 sows, 25 days after insemination, to check the accuracy of E₁C estimation as a test for early pregnancy diagnosis. Seventy-six of the examined animals were correctly diagnosed, giving an overall accuracy of 90.4%. No relationship was found between litter size and urine E₁C levels.     

Blastocyst is the source of prostaglandins in the implantation site in the rabbit

The levels of prostaglandins (PGs) were measured by radioimmunoassay in the interimplantation and the implantation sites as well as in the implantation site without the blastocyst in the rabbit on day 7 of pregnancy (168h $\text{post coitum}$). The concentrations of PGs were also determined in the blastocyst (PGF:101.59_+?4.33 and PGE-A:29.74_+?3.11 ng/blastocyst, n=6) and the blastocel fluid (PGF:253.55_+?39.56 and PGE-A:83.29_+?6.60 ng/100 ul, n-4) on day 7. The levels of both PGF and PGE-A were significantly higher in the implantation site as compared to interimplantation site (PGF:73.63±6.68 vs. 0.59±0.21 and PGE-A:25.52±3.30 vs. 1.22±0.18 ng/100 mg wet weight n=8). The removal of the blastocyst from the implantation site drastically reduced the concentrations of PGs in this site (PGF:8.71±2.80 and PGE-A:1.64±0.12 ng/100 mg wet weight, n=8). The results provide evidence that the blastocyst is the major source of PGs which contribute to the hige concentration in the implantation site in the rabbit.     

Uptake and effects of ovarian steroids in the early pig embryo: In vitro and in vivo studies

The role of ovarian steroids in the preimplantation pig embryo was studied in vivo and in vitro. Twenty gilts were treated three times daily on days 1 to 4 after insemination with either 25, 100, 250, or 1000 mg progesterone in oil, and 17 gilts were injected with corresponding amounts of sesame oil (controls). All gilts were slaughtered 5 days after insemination and the embryos were recovered. Oviduct and plasma progesterone content were significantly (P<0.001) higher in gilts treated with 750 mg of exogenous progesterone per day. After 750 mg progesterone, oviduct progesterone content was twice as high as control levels, while after 3000 mg progesterone per day the levels in oviduct and uterus exceeded those of controls by five and seven times, respectively. In gilts treated with 750 mg progesterone per day, plasma progesterone levels were 177.4 ± 22.1 ng/ml ($\text{x}\text{±}\text{SD}$) on day 3 and 186.4 ± 69.2 ng/ml on day 5 and resembled values found in superovulated pigs with more than 40 ovulations. Excessive plasma progesterone values of 1014.6 ± 840.4 ng/ml on day 3 and 473.2 ± 197.2 ng/ml on day 5 were found after treatment with 3000 mg of progesterone per day. Treatment with up to 750 mg of exogenous progesterone per day, did not affect embryonic development, but 3000 mg per day resulted in a significantly (P<0.001) higher percentage of retarded and degenerate embryos compared to controls (71.8% versus 3.2%).In addition, the amount and specificity of uptake of ³H-labelled progesterone and estradiol-17 beta by pig blastocysts recovered from superovulated gilts were investigated after 6 hrs in vitro culture. The uptake of ³H-progesterone was 131.9 ± 56.9 counts per million (cpm) per 10 blastocysts, corresponding to 1.1 fmoles progesterone. The uptake was non-specific for it was only slightly reduced in the presence of a 100-fold excess of unlabelled progesterone (20.1%) or estradiol-17 beta (27.0%). The uptake of ³H-estradiol-17 beta was 133.9 ± 74.12 cpm per 10 blastocysts, corresponding to 1.3 fmoles estradiol-17 beta. The uptake was significantly (P<0.01) reduced by 67.7% in the presence of a 100-fold excess of unlabelled estradiol-17 beta. Apparent specific binding was 0.87 fmoles estradiol-17 beta per 10 blastocysts or 72.5 fmoles estradiol-17 beta per mg protein. The uptake was only slightly reduced in the presence of a 100-fold excess of unlabelled progesterone (23.3%). This significant inhibition could be determined after 2 hrs in vitro culture. There was no competitive inhibition after 20 min. of culture.Uptake by unfertilized ova and degenerate embryos recovered on day 5 was significantly smaller (51.8 ± 10.3 cpm per 10 eggs; P<0.001) than by blastocysts recovered on the same day. No competitive inhibition could then be determined. In vivo, preimplantation pig embryos seem to be rather insensitive to high progesterone levels. Excessive amounts of progesterone probably can be metabolized to a great extent. Progesterone seems to be taken up rather non-specifically by the pig embryo. The uptake and binding of estradiol-17 beta seems to be more specific. Studies are in progress to investigate the physiological role of estradiol-17 beta uptake in early embryonic development.     

The extrasplanchnic origin of blood dihydrotestosterone in elderly men

The splanchnic extraction and interconversion of testosterone (T) and dihydrotestosterone (DHT) were studied in 5 elderly men undergoing cardiac catheterization using a constant Infusion of [1,2-³H] testosterone and [4-¹⁴C] DHT. Metabolic clearance rate (MCR), splanchnic extraction (SE), splanchnic clearance (SC), extrasplanchnic clearance (ESC), transfer constant In blood ([P]_BB^T-DHT) and transfer constant across the liver ([P]_BB^T-DHT) were calc?ulated. The MCR^T was 675 ± 108 (mean ± SC) L/day and MCR^DHT was 409 ± 68 L/day. SE^T was 45.9 ± 7.0% and SE^DHT was 18.5 ± 5.4%. When these values are compared with those recently reported by us for normal men, there is a $\text{1}\text{3}$ reduction in SE^T and $\text{1}\text{2}$ reduction for SE^DHT in elderly men. The calculated SC^T and ESC^T were 355 ± 72 L/day and 320 ± 86 L/day, respectively. SC^DHT and ESC^DHT were 145 + 48 L/day and 263 ± 77 L/day respectively, suggesting that a major fraction of DHT is metabolized in extrasplanchnic organs. No evidence for a net appearance of DHT by either mass or specific activity analysis in hepatic vein blood was observed indicating that the splanchnic compartment does not contribute DHT into the circulation either by $\text{de novp}$ synthesis or via conversion from testosterone. This work indicates that conversion of testosterone to DHT in elderly men occurs entirely in extrasplanchnic tissue.     

Effects of phyto-estrogenic alfalfa consumption on plasma LH levels in cycling ewes

Plasma luteinizing hormone (LH) concentrations were determined in five Dorset ewes fed orchard grass hay (Dactylus glomerata) and five ewes fed alfalfa (Medicago sativa). Total phyto-estrogen content ($\text{X}\text{±}\text{SEM}$ genistein equivalents) of the orchard grass hay and alfalfa was 16.9 ± 2.9 and 118 ± 12.3 ppm respectively. LH was determined at regular intervals during the estrous cycles synchronized with progesterone impregnated pessaries and characterized by marker ram and vaginal cytology.Peak LH levels in control ewes (40.1 ± 5.5 ng/ml) were lower (P<0.05) than in ewes fed phyto-estrogenic alfalfa (66.0 ± 16.8 ng/ml). Results also indicate that the LH peak may occur later (P<0.05) in the estrus period of ewes fed phyto-estrogenic alfalfa (15.4 ±4.5 h). These experiments may suggest that peak LH concentrations are elevated and delayed further into the estrus period in ewes fed phyto-estrogenic alfalfa.     

The aging leydig cell: VII. Cytoplasmic estradiol receptors

Plasma estradiol and cytosolic estradiol receptor levels of testes were determined in a group of young (2–3 months) and old (24 months) Sprague-Dawley rats. Estradiol binding sites for the young rats averaged 5.6 ± 0.3 fmol/mg protein ($\text{x}\text{±}\text{SE, n}\text{=12}$), which was comparable to that of the old rats, 5.7 ± 0.3 fmol/mg protein (n=12). Using Scatchard analyses, the association constants at equilibrium of estradiol receptor binding of the old and young rats were the same, 6.1 × 10¹⁰M^?1. Plasma estradiol levels were also similar in both groups-19.6 ± 2.8 pg/ ml (n=14) for the young and 19.2 ± 2.6 pg/ml (n=10) for the old rats. Our results suggest that impaired testosterone biosynthesis in old rats was not due to elevated plasma estradiol levels or to differences in testicular estradiol receptor content.     

Plasma prolactin in the periparturient sow

A homologous radioimmunoassay was used for measurement of porcine prolactin in blood plasma collected from sows during the periparturient period. The assay was able to detect prolactin over a range of 0.5 to 7.0 ng/assay tube. There was no significant cross reaction with growth hormone, luteinizing hormone, or follicle stimulating hormone at amounts up to 10⁵ ng/assay tube while porcine ACTH gave 30% binding at 10⁴ ng. Prolactin was not detected in plamsa from a hypophysectomized pig or 2 ergocryptine-treated sows when 100 μ l plasma were assayed. Prolactin concentration in plasma was then measured in 14 periparturient sows within a period extending from 7 days before farrowing to 7 days after farrowing. Samples were collected at 15 min intervals between 1330 and 1630 h each day. However, prolactin assays were done only on the even-numbered samples (30 min interval). Plasma prolactin concentrations (ng/ml, $\text{X}\text{± SEM}$) were 23.7 ± 2.0 on days ?7 to ?5 prepartum, began to rise by day ?3 prepartum (42.5 ± 5.9), and peaked at 127.5 ± 17.6 on day 1 prepartum. By day 3 postpartum, prolactin concentrations in plasma had decreased to 80.5 ± 12.6 and further declined to 51.6 ± 4.6 on day 7 postpartum. The mean prolactin concentration in plasma for all pigs on days ?1 to +2 was 116.8 ± 13.8. This mean concentration for days ?1 to +2 was different (P < 0.025) from the mean prolactin concentration for the period both prior and subsequent to these days (?8 to ?2 and +3 to +8 days).     

Effects of lighting programs on onset of the ovulatory season in mares

Using 240 pony mares, lighting regimens were tested for their efficiency in hastening the onset of the ovulatory season. The mean number of days from January 1 to first ovulation was used as the end point. No advantage was gained by beginning a fixed lighting regimen ($\text{15.5L}\text{8.5D}$, hours light/hours dark) November 1 (66 ±8) versus December 1 (65 ±9), but beginning on January 1 was less efficient (98 ±8; controls, 132 ±5; P<0.05). In another experiment, daily three-hour interruptions of either the light phase (67 ±10) or the dark phase (71 ±11) did not significantly retard the effectiveness of a fixed regimen of $\text{15L}\text{9D}$ (54 ±5; controls, 142 ±6). A $\text{15L}\text{9D}$ regimen every other day (natural day length on alternate days) resulted in an interval (85 ±7) that was shorter (P<0.05) than for the controls and longer (not significant) than for the daily $\text{15L}\text{9D}$ regimen. When used with natural day length, a one-hour pulse of light in the evening (15 hours after sunrise) was not effective (141 ±6); a one-hour pulse in the morning 9.5 hours after sunset) was only partially effective (117 ±6). In another experiment, the interval was reduced (P<0.05) in a group with one hour of light fixed at 4:00 a.m. with natural day length (85 ±8; $\text{15L}\text{9D}$, 75 ±7; controls, 126 ±9). Results indicated that a fixed one-hour pulse of light at 4 a.m., used with natural day length, may provide an acceptable level of stimulation.     

Daily sperm production of zebus (Bosindicus) estimated by quantitative histology of the testis

Microscopic parameters were studied quantitatively in testes from six Nelore zebus ($\text{Bos}$$\text{indicus}$), aged 4 to 6 years, with normal spermatogenesis, which were kept at sexual rest. Ratios of germ cell nuclei, counted in cross sections of seminiferous tubules, indicated that cellular losses in the spermatogenic process of the zebu are higher than those observed in taurines. Daily sperm production, estimated from the number of round spermatids in stage 1 of the cycle of the seminiferous epithelium and the duration of this cycle, was (mean ± SD) 12.2 ± 0.9 × 10⁶ spermatids/gram of testis parenchyma/day and 2.6 ± 0.5 × 10⁹ spermatids/testis/day. These values are smaller than those of taurines.     

A direct radioimmunoassay for 11-deoxycortisol

A rapid, sensitive and highly specific radioimmunoassay (RIA) for 11-deoxycortisol has been developed and used for the measurement of serum concentrations.Antisera were raised using 11-deoxycortisol-3-carboxymethyloxime-bovine serum albumin as immunogen and showed minimal cross reaction with related steroids. 11-Deoxycortisol-3-carboxymethyloxime was coupled to ¹²⁵I-iodohistamine to produce a labelled antigen of high specific activity (s.a. 680 Ci/mmol). The assay was performed for 1h at room temperature and pH 4.Results were correlated with those after extraction, high pressure liquid chromatography and RIA of serum samples (y = 0.78x + 36.9; r = 0.97, p<0.001). The accuracy of the method was satisfactory (y = 1.00x ? 0.61; r = 0.95; p<0.001). Assay sensitivity was 0.3 nmol/1. 11-Deoxycortisol concentrations in normal subjects at 09.00h were 26 ? 46 nmol/1 $\text{(37.2 ± 5.7;}\text{x}\text{± 1}\text{S.D.}\text{)}$.The assay should facilitate the investigation of patients with possible abnormalities of adrenocortical function.     

Sulfation of gastrin in Zollinger-Ellison sera: evidence for association between sulfation and proteolytic processing

Sulfated gastrins were quantitated in sera from 15 patients with the Zollinger-Ellison syndrome (ZES) by specific radioimmunoassays. The total concentration of gastrin varied from 174 to 285000 pmol/1. Sulfated gastrins constituted $\text{44.8±5.5\%}$ (mean ± S.E.M.) of the gastrins in ZES sera compared with $\text{37.7±1.9\%}$ in sera from 100 control subjects ($\text{P>0.1}$). There was no correlation between gastrin concentration and sulfation ($\text{r=0.40}$). Gel and ion-exchange chromatography showed that up to 90% of the gastrins could be in the sulfated form. The highest degree of sulfation was found in sera where the small gastrin components dominated. Thus, the percentage of small gastrins (G-17 and G-14) correlated with the degree of sulfation ($\text{N = 15}$, $\text{r=0.75}$, $\text{P<0.01}$). We suggest therefore that proteolytic processing of the gastrin precursor and sulfation of tyrosyl are associated.     

An ultrastructural examination of everted rat jejunum

Male, albino, Sprague-Dawley rats were sacrificed by cervical separation. Segments of jejunum were excised, everted and examined with the electron microscope. Examination of tissue fixed immediately after eversion revealed the following changes as compared to non-everted segments fixed $\text{in}$$\text{situ}$ and $\text{in}$$\text{vitro}$: 1) an increase in the length of microvilli from (mean ± S. E.) 0.991 ± 0.011μ for normal tissue to 1.389 ± 0.023μ for everted tissue, 2) an increase in width of microvilli from (mean ± S. E.) 0.089 ± 0.001μ for normal tissue to 0.097 ± 0.001μ for everted tissue, 3) an increase in length and number of lateral membrane interdigitations, and 4) the appearance of intercellular “lakes” in the lateral spaces. The above changes are in those structures hypothesized to be involved with salt and water transport across epithelia and may reflect altered transport rates $\text{in}$$\text{vitro}$ as compared to $\text{in}$$\text{vivo}$.     

N-acetylation of dopamine and tyramine by mosquito pupae (Aedes togoi)

The $\text{N-}\text{acetyltransferase}$ (NAT) activity of mosquito pupae was measured by a radioenzymatic assay, using [¹⁴C]-, [³H]dopamine, [¹⁴C]tyramine or [¹⁴C]acetyl-CoA. The pupal extract could also generate acetyl-CoA from ATP, acetate and CoA for this acetylation reaction. Both the dopamine- and tyramine-NAT reactions proceeded linearly up to 20 min at an optimum pH of 8.4. It is possible that the same enzyme is involved in the acetylation of both biogenic amines as shown by the competitive inhibition kinetics obtained, and the similarities of the NAT reaction with both amines, in the presence of metal chelators, metal ions, SH reagents and MAO inhibitors. Mn²⁺ stimulated and Zn²⁺ inhibited the reaction. The specific activity of NAT in individual pupae measured soon after pupation showed no significant difference between the male and female pupae: the values obtained were, respectively, $\text{893 ± 57}$ and $\text{861 ± 30}$ pmol [¹⁴C]NAcT formed/min per mg protein and $\text{21.9 ± 1.2}$ and $\text{22.0 ± 1.4}$ pmol [³H]NADA formed/min per mg protein.     

Fertility of high-yielding dairy cows following superovulation and non-surgical recovery of embryos

It was the aim of this investigation to study the combined effect of superovulation and non-surgical recovery on the fertility of the donor animals. Injection of a prostaglandin analogue at the day of collection (days 6–8 after standing heat), significantly shortened the super-ovulatory estrus cycle (21.3 ± 10.0 days), when compared with untreated donor animals (38.7 ± 15.1 days). The prostaglandin treatment, however, also led to more irregular estrous cycles, resulting in a lower pregnancy rate after first insemination (47%) and a need for more inseminations per conception (2.00 ± 1.00), than in animals not treated with prostaglandin analogue (92.3% and 1.06 ± 0.29 AI/conception). For all animals these parameters were 66.6% and 1.60 ± 0.99 AI/conception). The average time from calving to pregnancy was 143.1 ± 34.3 days, slightly longer for prostaglandin treated (148.2 ± 39.4 days) than for untreated animals (137.3 ± 22.8 days). One animal developed endometritis and one had adhesions in the $\text{bursa ovarii}$. It was found that most donors attained normal fertility, and that superovulation was more likely to affect the fertility (abnormal cyclicity, early embryonic mortality e.g.) than the flushing of the uterus. Treatment with a prostaglandin analogue at the day of collection did not improve the subsequent fertility.     

Greatly extended viability of rat brain storage vesicles in an intracellular medium based upon a non-permeant polyanion

The MgATP-stimulated accumulation of (-)-³H-nor- epinephrine (NE) by rat brain neuronal storage vesicles has been characterized in a new medium based upon polyacrylic acid (avg. MW 5,000). The medium allows careful regulation of K⁺ concentration (140 mM), has a large buffer capacity, and is non-permeant to membranes. Light scattering measurements have confirmed the osmotic stability of vesicles suspended in this medium. Vesicular accumulation of (-)-³ H-NE (Km 1 × 10^?6 M) in this system (37°) was examined under saturating (10^?5 M) and non-saturating (2 × 10^?7 M) concentrations of NE. At 10^?5 M NE, uptake saturated at 5 min and remained stable for periods up to one hour, with maximal uptake levels (pmol/mg protein) of 15.7±0.30 (37°), 3.0±0.49 (0°), 4.4±0.22 (reserpine pretreated $\text{in}$$\text{vivo}$) and 6.0±0.79 (without MgATP). At 2×10^?7 M NE uptake was biphasic with maximal uptake levels (pmol/mg protein) of 4.04±0.14 (37°), 0.19±0.01 (0°), 0.95±0.01 (reserpine) and 0.83±0.08 (without MgATP). Vesicle preparations refrigerated in this medium for 24 hrs displayed properties quite similar to those measured acutely (NE = 2.2x10^?7 M).     

Weak acid accumulation in the serosal extracellular compartment of the frog gastric mucosa

The dimethyloxazolidine dione distribution in the extracellular compartments of the frog gastric mucosa was analyzed by washout kinetics. The volumes of the two extracellular compartments, serosal and mucosal, were estimated by inulin washout as $\text{0.435 ± 0.019}$ and $\text{0.176 ± 0.018 μl/μl}$ tissue water, respectively. In the serosal extracellular space, significant dimethyloxazolidine dione accumulations of $\text{2.63 ± 0.25}$, $\text{2.28 ± 0.16}$, and $\text{1.86 ± 0.08}$ times that of the bathing media were found for bathing solutions with pH values of 6.9, 7.4, and 7.9 respectively. A high pH of the serosal extracellular fluid by itself could not account for the high values of dimethyloxazolidine dione accumulation. A difference in the total dimethyloxazolidine dione accumulation requires: (a) the existence of differences in the pH values and also the existence of a difference in the diffusion coefficient of the two forms of dimethyloxazolidine dione; or (b), a binding of one of the two forms, i.e., binding of dimethyloxazolidine dione form by fixed charges.