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Biomedical subjects

J M Fernandez

Publications and source records attributed to J M Fernandez.

At least 91 records · Page 5Linked to original sources

Dietary protein and energy restriction in mares: rapid changes in plasma metabolite and hormone concentrations during dietary alteration.

Two diets consisting of bermudagrass hay and a corn-cottonseed hull-based supplement were formulated to provide either 100% (control) or 50% (restricted) of the protein and energy requirements for maintenance for mature mares. Twelve light horse mares were fed the control diet for 7 d, and then at 0800 on d 0, six mares were switched to the restricted diet. All diets were fed as two equally sized meals at 0800 and 1600. At 0800 on d 7, mares receiving the restricted diet were switched back to the control diet. Relative to control mares, mares switched to the restricted diet had reduced plasma concentrations of glucose (P = .005) and insulin (P = .09) in response to the two restricted meals on d 0. However, concentrations of both glucose and insulin returned to control levels (P > .1) within 1 h after the consumption of the control diet on d 7. Dietary restriction increased (P = .009) plasma NEFA concentrations within the first 24 h, and NEFA concentrations remained elevated (P < .001) in restricted mares until the mares were returned to the control diet on d 7. Meal-induced increases (P < .05) in plasma concentrations of glucose, insulin, urea N, glucagon, and thyroxine were observed. These results emphasize the importance of early sampling when monitoring plasma constituents during nutrient alterations and indicate that prefeeding responses of plasma constituents alone may not fully explain the metabolic consequences of nutrient restriction.

Analysis of Variance↗

Seasonal effects of supplemental fat or undegradable protein on the growth and metabolism of Holstein calves.

One summer (heat stress) and one winter (neutral conditions) trial were conducted to assess the effects of varied amounts of RUP and fat on the growth and metabolic responses of calves. In each trial, 190-kg Holstein calves (n = 45) were fed a basal diet of corn and cottonseed hulls supplemented with soybean meal (control) or supplemented with raw or roasted whole soybeans or the basal ration plus prilled hydrogenated tallow either with or without a mixture of feather meal and blood meal. Prilled tallow was more detrimental to fiber digestion and growth performance than were whole soybeans, especially during summer. During winter, calves fed a high (49%) percentage of the dietary CP as RUP had similar BW gain and efficiency of feed usage as did calves fed a moderate percentage (39%), but, during summer, calves fed the high percentage of RUP had greater BW gain and efficiency than did calves fed moderate percentages of RUP. Ruminal concentrations of VFA were parallel to digestibilities of fiber in the total tract. Ruminal concentrations of ammonia or blood concentrations of urea were not consistent with the concentrations of RDP fed. Concentrations of glucose, NEFA, insulin, and thyroid hormones in the blood were primarily affected by season. Data suggested that a higher percentage and AA quality of RUP improved the growth of calves during summer but not during winter.

Ammonia↗

Colocalization of calcium entry and exocytotic release sites in adrenal chromaffin cells.

"Snapshot" images of localized Ca2+ influx into patch-clamped chromaffin cells were captured by using a recently developed pulsed-laser imaging system. Transient opening of voltage-sensitive Ca2+ channels gave rise to localized elevations of Ca2+ that had the appearance of either "hotspots" or partial rings found immediately beneath the plasma membrane. When the Ca2+ imaging technique was employed in conjunction with flame-etched carbon-fiber electrodes to spatially map the release sites of catecholamines, it was observed that the sites of Ca2+ entry and catecholamine release were colocalized. These results provide functional support for the idea that secretion occurs from "active zone"-like structures in neuroendocrine cells.

Adrenal Medulla↗

Increased messenger RNA expression of the 695 and 751 amino acid isoforms of the beta-amyloid protein precursor in the thalamus of 17-year-old cynomolgus (Macaca fascicularis) monkeys.

The levels of expression of messenger RNAs of the 695 and 751 amino acid isoforms of the beta-amyloid protein precursor in the brains of three-year-old and 17-year-old cynomolgus monkeys (Macaca fascicularis) were visualized and quantified by in situ hybridization histochemistry using 35S-labelled oligonucleotide probes. The analysis was carried out on coronal brain sections taken through the hippocampus and thalamus at the level of the geniculate nuclei. High densities of beta-amyloid protein precursor695 and beta-amyloid protein precursor751 messenger RNAs were found in the medial aspects of the mediodorsal, centromedian and parafascicular nuclei of the 17-year-old monkeys. The messenger RNA levels of the 695 and 751 isoforms were about two- and seven-fold, respectively, those found in the same nuclei of the three-year-old animals. The levels of these messenger RNA transcripts in the 17-year-old monkeys were not significantly different from those in the three-year-old animals in other brain areas e.g. the temporal cortex, entorhinal cortex and hippocampus. No Alzheimer's disease-like neuropathology in terms of diffuse or senile beta-amyloid plaques, dystrophic neurites or neurofibrillary tangles were detectable by specific innumohistochemical procedures in the above thalamic nuclei of the 17-year-old animals. In addition no reactive gliosis was seen in the thalamus of these monkeys.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Do caged-Ca2+ compounds mimic the physiological stimulus for secretion?

We measured the exocytotic response induced by a rapid and global increase in the concentration of Ca2+ or GTP gamma S (achieved by flash photolysis of caged compounds) in mast cells and chromaffin cells. Secretion was measured by following both cell membrane capacitance and amperometry. When Ca2+ was used to trigger secretion, we observed an immediate increase in capacitance, however, the first amperometric spike observed was delayed with respect to the change in capacitance (approximately 5 s in mast cells and approximately 0.6 s in chromaffin cells). In contrast, when GTP gamma S was used to trigger secretion, no such discrepancy was seen, both measurements reported a secretory response that was similarly delayed with respect to the stimulus. These data suggest that the capacitance increase, when triggered by a large Ca2+ step, reports events that do not result in the fusion of vesicles containing oxidizable substances. These results contrast with the smaller secretory responses evoked by the transient opening of Ca2+ channels. Under these more physiological conditions, pulsed-laser imaging studies revealed that the Ca2+ influx was localized to discrete areas at the plasma membrane. Thus the Ca2+ stimulus triggered by DM-nitrophen fails to reproduce the small localized changes seen during a physiological stimulation and creates an artifactual non-secretory capacitive response.

Animals↗

Hydrophobic ions amplify the capacitive currents used to measure exocytotic fusion.

The detection of exocytotic fusion in patch-clamped secretory cells depends on measuring an increase in the cell membrane capacitance as new membrane is added to the plasma membrane. However, in the majority of secretory cells, secretory vesicles are too small (< 200 nm in diameter) to cause a detectable signal. We have found that incubations of normal mouse mast cells with the hydrophobic anion dipicrylamine (DPA), increases cell membrane capacitance by about three times. The large capacitive current induced by DPA was voltage-dependent, having a maximum value at -10 mV. The DPA-induced charge movement could be described by a single barrier model in which the DPA molecules move between two stable states in the bulk lipid matrix of the membrane. More importantly, the DPA treatment produced a sevenfold increase in the size of the capacitance steps observed upon the exocytotic fusion of single secretory granules. A similar amplification of DPA on the secretory vesicle capacitance was observed in a cell with larger (< or = 5 microns in diameter) or with smaller secretory granules (< 250 nm in diameter). Additionally, the increased granule membrane capacitance enlarged the transient capacitive discharge measured upon formation of a fusion pore in normal mast cell granules. Our results indicate that hydrophobic ions provide an important tool for high resolution studies of membrane capacitance.

Animals↗

The secretory granule matrix-electrolyte interface: a homologue of the p-n rectifying junction.

When placed at the tip of a glass micropipette electrode the polymeric matrix of the secretory granule behaves like a diode. The measured current was 100-fold greater at negative potentials compared to positive potentials, and up to sixfold greater than that measured with the pipette alone. By manipulating the geometry of the electric field we show that these electrical properties result from focusing an electric field at the gel-electrolyte interface. We also show, by using pulsed-laser imaging with fluorescein as the ionic probe, that there is a rapid accumulation and depletion of ions at the gel-electrolyte interface. A voltage pulse of -9 V applied to the gel caused a severalfold increase in the fluorescence intensity within 5 ms. This correlated with an increase in the measured current (approximately 1 microA). In contrast, within 5 ms of applying +9 V we recorded a decrease in the fluorescence intensity, which paralleled the twofold decrease in the measured current. This is similar to a p-n junction where an applied voltage causes the accumulation and depletion of charge carriers. Using synthetic gels (diameter 3-6 microns) with different charge characteristics we observed no rectification of the current with neutral gels and confirmed that rectification and amplification of the current were dependent on the fixed charge within a gel. In addition, we modeled the conduction at the gel-electrolyte interface using the Nernst-Planck electrodiffusion equation and accurately fitted the experimental current-voltage relationships. This study provides some insight into how biological interfaces may function. For example, we suggest that neurotransmitter release during exocytosis could be regulated by voltage-induced accumulation and depletion of ions at the interface between the secretory granule and the fusion pore.

Animals↗

Dietary protein and(or) energy restriction in mares: plasma glucose, insulin, nonesterified fatty acid, and urea nitrogen responses to feeding, glucose, and epinephrine.

Sixteen light horse mares (8 to 9 yr of age; 457 to 579 kg BW) were fed Bermudagrass hay and a corn/cottonseed hull-based supplement formulated to contain either 100% (control) or 50% (restricted) of the protein and(or) energy requirements for maintenance in a 2 x 2 factorial arrangement of treatments. Daily measurements of intake, BW, and plasma hormones and metabolites were made for 33 d. Plasma glucose, insulin, NEFA, and urea N were measured in hourly samples drawn on d 27, and parallel with an i.v. glucose tolerance test (IVGTT) and epinephrine challenge on d 29. Energy restriction increased daily NEFA concentrations (P < .001) and urea N (P = .013), whereas protein restriction decreased (P = .002) urea N concentrations. These effects of protein and energy restriction occurred within 24 h and were consistent (day effect, P > .1) throughout the remaining 24 d. Normal meal consumption elevated plasma glucose, insulin, and urea N concentrations (time effect, P < .08). Plasma NEFA concentrations did not change after feeding in mares fed control energy, but decreased in mares fed restricted energy (energy x time interaction, P = .005). After IVGTT, areas under the curve for plasma glucose and insulin were smaller in mares fed restricted protein (P < .05), whereas glucose area was larger in mares fed restricted energy (P = .009). After epinephrine injection, energy restriction increased the initial magnitude of the NEFA response, but after 50 min, reduced plasma NEFA below pre-injection concentrations (energy x time interaction, P = .06).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Technical note: tissue residues of mimosine and 2,3-dihydroxypyridine after intravenous infusion in goats.

Sixteen growing Alpine wethers (average BW 35 +/- 2 kg) were assigned to one of four treatments to evaluate tissue retention of the leucaena toxins mimosine (MIM) and 2,3-dihydroxypyridine (2,3-DHP). Treatments were infused i.v. for 2 d and were 1) saline control, 2) MIM (200 mg.kg BW-.75.d-1), 3) 2,3-DHP (200 mg.kg BW-.75.d-1), or 4) MIM (100 mg.kg BW-.75.d-1) + 2,3-DHP (100 mg.kg BW-.75.d-1). Immediately after the infusion, the goats were slaughtered and tissue concentrations of MIM and 2,3-DHP were determined via HPLC. No detectable levels of either toxin were found in spleen, heart, lung, or muscle; however, appreciable amounts of MIM and 2,3-DHP were found in plasma, kidney, and liver samples. Kidney MIM content was greater (P < .01) than that of liver, although liver tended to retain slightly more 2,3-DHP (P > .05). Infusion of MIM resulted in a plasma MIM content of 39 to 54 mumol/L and reduced (P < .01) plasma PHE and LEU. Infusion of 2,3-DHP resulted in a plasma 2,3-DHP content of 9.4 mumol/L and increased plasma THR, ARG, VAL, PHE, ILE, LEU, and LYS concentrations (P < .10). Humans consuming offals from ruminants consuming large amounts of the leguminous forage leucaena may be exposed to appreciable quantities of MIM and 2,3-DHP.

Animals↗

Endocrine and metabolic response to muscarinic stimulation and inhibition in the ruminant: effects of slaframine.

The influence of slaframine (SF), a parasympathomimetic compound isolated from the fungus Rizoctonia leguminicola, on circulating metabolic hormone concentrations was investigated in goats. In Exp. 1, SF was administered i.v. at 0 (CONT), 50 (LSF), 100 (MSF), or 150 (HSF) microgram/kg.75 BW in four mature Spanish-cross does (average BW 36 +/- 7 kg) fitted with indwelling jugular vein catheters in a 4 x 4 Latin square design. Plasma glucose peaked (P < .06) at 120 min with LSF and at 180 min with HSF and was higher (P <.06) than the CONT at these times. Glucose exhibited a quadratic response (P < .03) to SF. Area under the response curve for glucose differed (P < .02) in HSF from CONT and MSF. Insulin peaked (P < .01) at 240 min with MSF and at 180 min with HSF. Plasma triiodothyronine was maintained at a higher level (P < .03) with HSF. Thyroxine peaked (P < .06) at 120 min with MSF and 300 min with HSF. Plasma NEFA and somatotropin concentrations were not affected (P > .10) by SF. In Exp. 2, four mature Spanish-cross wethers (average BW 27 +/- 2 kg) fitted with jugular vein catheters were administered SF (0 and 114 micrograms/kg.75 BW) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4DAMP; 0 and 258 micrograms/kg.75 BW), a M3-muscarinic receptor antagonist, i.v. in a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. With SF, glucose peaked (P < .06) at 60 min and insulin peaked (P < .05) at 180 min. Plasma triiodothyronine levels were maintained (P < .05) with SF but declined with other treatments. Plasma NEFA and thyroxine concentrations remained unchanged regardless of treatment. Slaframine administration induced hyperglycemia and hyperinsulinemia in goats; however, these changes were blocked by preadministration of isomolar quantities of the M3-muscarinic receptor antagonist, 4DAMP.

Alkaloids↗

Feed deprivation of mares: plasma metabolite and hormonal concentrations and responses to exercise.

Twelve light horse mares were fed a control diet that provided 100% of their maintenance protein and energy requirements for 7 d and were then either continued on the control diet or totally deprived of feed (with access to water) for 3 d . Plasma samples were drawn twice daily throughout the experiment, at 15-min intervals for 9 h beginning 45 h after feed removal, and at 10-min intervals around an exercise bout beginning 73 h after feed removal. Feed deprivation increased (P < or = .06) whole blood beta-hydroxybutyrate and plasma NEFA, urea N, L-lactate, and glucagon concentrations, decreased (P = .02) IGF-I concentrations, and did not change (P > .1) plasma glucose insulin, prolactin, triiodothyronine, and thyroxine concentrations. Exercise increased (P < .05) plasma NEFA, prolactin, and growth hormone (GH) concentrations in all mares. Plasma NEFA concentrations increased (P < .001) after exercise and remained increased in fed mares, but rapidly decreased in deprived mares (time x diet interaction, P = .006). Plasma glucose concentrations following exercise increased in deprived mares but decreased in fed mares (time x diet interaction, P = .07). The plasma prolactin response after exercise also differed between groups (P = .09). Feed-deprived mares had greater (P = .02) plasma GH concentrations before exercise (73 h after feed withdrawal) and had a greater (P < .001) GH peak at 10 min after initiation of exercise. The increase in secretion rate o GH due to feed deprivation in these mares was similar to that reported for other domestic species but was not nearly as great in magnitude.

Animals↗

Effect of chromium tripicolinate on growth, glucose tolerance, insulin sensitivity, plasma metabolites, and growth hormone in pigs.

Two experiments were conducted to evaluate the effect of Cr as Cr tripicolinate (CrPic) on growth, glucose tolerance, insulin sensitivity, plasma metabolites, and growth hormone (GH) in pigs. Pigs were fed a control diet or a diet supplemented with 200 micrograms of Cr/kg of diet as CrPic. Thirty (15 per diet, initial BW was 21.3 kg) and 24 (12 per diet, initial BW was 24.9 kg) crossbred barrows were used in Exp. 1 and 2, respectively. The diets were formulated to provide 120% of the lysine requirement for 20- to 50-kg pigs. A glucose tolerance test (IVGTT; 500 mg of glucose/kg BW) and an insulin challenge test (IVICT; .1 IU of porcine insulin/kg BW) were conducted. In addition, during Exp. 1, a GH-releasing hormone (GHRH) challenge was conducted. All data were pooled across experiments, except where noted. Average daily gain, ADFI, gain/feed, and fasting (15 to 18 h) plasma glucose and total protein concentrations were not affected (P > .10) by dietary treatment. Fasting plasma cholesterol (P < .05) was increased and NEFA (Exp. 2 only, P < .02), urea N (P < .07), and insulin (P < .10) concentrations were decreased in pigs fed CrPic. During the IVGTT and IVICT, glucose disappearance rate (k, percentage/minute) was increased (P < .04) and glucose half-life (t1/2, minutes) was decreased (P < .04) in pigs fed CrPic; however, insulin kinetics were not altered (P > .10). During the GHRH challenge, pigs fed CrPic had decreased (P < .09) area under the response curve for GH.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Dietary protein and(or) energy restriction in mares: plasma growth hormone, IGF-I, prolactin, cortisol, and thyroid hormone responses to feeding, glucose, and epinephrine.

Sixteen light horse mares were fed diets of bermudagrass hay and a corn/cottonseed hull-based supplement formulated to contain either 100% (control) or 50% (restricted) of the protein and(or) energy requirements for maintenance in a 2 x 2 factorial arrangement of treatments. Plasma IGF-I, prolactin, cortisol, triiodothyronine, and thyroxine were monitored for 33 d. On the 27th d, frequent blood samples were drawn throughout the day for the measurement of growth hormone (GH), and on the 29th d, an epinephrine challenge and an i.v. glucose tolerance test (IVGTT) were performed in the morning and afternoon, respectively. Restriction of protein and(or) energy reduced (P < .001) plasma IGF-I concentrations within 24 h, and the effect persisted through the 24th d. Energy restriction decreased (P = .01) plasma cortisol concentrations, whereas thyroid hormones were not influenced (P > .1) by restriction of protein and(or) energy. Plasma prolactin concentrations were low throughout the experiment and after the IVGTT, but they increased (P = .003) after feeding. Protein restriction increased (P = .09) the occurrence of GH episodes during the 14-h feeding period on d 27; the greatest effect occurred in the mares restricted in both nutrients. In contrast, energy restriction reduced (P = .05) the GH response to epinephrine injection. We conclude that 1) protein deficiency in mares increases GH secretion, whereas energy restriction alone does not, 2) a deficiency in energy and(or) protein reduces IGF-I secretion, and 3) prolactin concentrations increase after feeding, even at a time of year when secretion rates are naturally low.

Animals↗

Influence of chromium tripicolinate on glucose metabolism and nutrient partitioning in growing lambs.

Twenty-four Suffolk lambs (average BW 38 +/- 2.7 kg; 16 ewes and 8 wethers) were fed either a corn-cottonseed hull-based control diet (CON) or CON plus 250 ppb of Cr as chromium tripicolinate (CrPic). Lambs were penned in groups of three and ADG and DMI were measured through d 85 of the experiment. Jugular blood samples were obtained during wk 2, 7, and 11. An i.v. glucose tolerance test (IVGTT; 500 mg of glucose/kg BW) and an i.v. insulin challenge test (IVICT; .1 IU of ovine insulin/kg BW) were performed during wk 2 and 10 of the trial. This was followed by a N balance trial during wk 3 and 11. Wethers were slaughtered at the end of the experiment and carcass characteristics determined. No differences (P > .10) were observed between dietary treatments in DMI, ADG, or N balance; however, the CrPic-fed wethers had 18% less fat over the 10th rib (P = .082) and a lower yield grade (P = .014). Plasma NEFA was lower throughout the trial (P < .03) and cholesterol was 17% lower during wk 2 (P < .02) in lambs fed CrPic. There were no differences due to diet (P > .10) in plasma concentrations of urea N, glucose, albumin, total protein, insulin, glucagon, triiodothyronine, or thyroxine. Glucose clearance rate and half-life during the IVGTT and IVICT did not differ (P > .10) between CON and CrPic groups; however, during the IVGTT on wk 2, plasma insulin was elevated (P < .05) and glucose reduced (P = .067) in the lambs fed CrPic. Supplemental CrPic seems to influence metabolic measurements that may affect performance of growing lambs.

Animals↗

Responses of early lactation cows fed winter and summer annual forages and undegradable intake protein.

Twenty-four multiparous Holstein cows were divided into six blocks of 4 cows based on their previous 305-d mature equivalent milk yield and were used in a 2 x 3 factorial experiment for the first 84 DIM. Objectives were to determine the effects of forage and RUP supplementation of DM and nutrient intakes and digestibilities, milk yield and composition, BW change, and plasma concentrations of insulin, triiodothyronine, thyroxine, cortisol, NEFA, urea N, and protein. One cow from each block was assigned to wheat or sorghum silage plus one of three concentrates. Protein treatments were 1) all supplemental CP from soybean meal; 2) 33.3% of supplemental CP from heat soybean meal, fish meal, and corn gluten meal; and 3) protein treatment 2 plus an additional 4.6% CP from soybean meal. Intakes of DM, CP, NDF, ADF, NEL, and RUP were higher in cows fed sorghum silage; RUP tended to increase DMI. Milk and 3.5% FCM yields were higher for cows fed sorghum silage than for those fed wheat silage (42.3 vs. 40.0 kg/d and 41.2 vs. 38.7 kg/d, respectively). Milk protein, lactose, and SNF were increased by RUP. Insulin was increased, cortisol was decreased, and thyroid hormones were unaffected by RUP. Concentrations of NEFA were highest at wk 4. For early lactation cows fed diets containing sorghum or wheat silage, RUP is a necessity.

Animal Feed↗

Brain lesions and water-maze learning deficits after systemic administration of kainic acid to adult rats.

The relationship between hippocampal damage and spatial learning deficiencies was studied in rats injected with kainic acid (10 mg/kg i.p.). A single injection was given either before or after the acquisition phase of the Morris water-maze task. In this acquisition phase, the animals were required to find a hidden underwater platform starting from four different points. The task was repeated twice a day for 10 days. In the retention phase after 10 days rest, the rats repeated the same task. The damage caused by the treatment occurred in several prosencephalic areas, including the piriform and enthorhinal cortices, the thalamus and the hippocampus. In the latter, greatest damage was seen in CA1 followed by CA3 while CA2 and the gyrus dentatus appeared almost unaffected. The behavioural results indicated that kainic acid impaired but did not preclude the acquisition of the water-maze task. During the retention phase, no significant differences in latencies were found between animals that were treated before and after acquisition, thus, indicating that pretraining does not play an important role in the recovery of these spatial abilities following hippocampal lesions.

Animals↗