Days Of Refeeding Research Articles

Two strategies for coping with food shortage in desert golden spiny mice

Desert rodents face periods of food shortage and use different strategies for coping with it, including changes in activity level. Golden spiny mice ( Acomys russatus) inhabit rock crevasses and do not dig burrows nor store food. When kept under 50% food restriction most, but not all, golden spiny mice defend their body mass by physiological means. We tested the hypothesis that these rodents use two different behavioral strategies, i.e., increasing activity level and searching for food or decreasing activity level and conserving energy to cope with food shortage. Twelve golden spiny mice were fed ad libitum for 14 days, followed by 40 days of 50% food restriction, and 14 days of refeeding. Body mass, food consumption and general activity were monitored. Seven mice significantly reduced activity level, concentrating their activity around feeding time, lowering energy expenditure and thus keeping their body mass constant (“resistant”), while five (“non-resistant”) significantly increased activity level (possibly searching for food) and thus energy expenditure, thereby losing mass rapidly (more than 25% of body mass). The non-resistant golden spiny mice were active throughout many hours of the day, with high variability both between and among individuals. The use of two strategies to cope with food shortage as found in the golden spiny mice may be of evolutionary advantage, since it allows a more flexible reaction to food restriction at the population level.

Coordinated regulation of the GH/IGF system genes during refeeding in rainbow trout (Oncorhynchus mykiss)

The GH/IGF system is a complex regulation network strongly dependent on nutrient availability. While the effect of starvation on the GH/IGF system has been extensively studied, the time course of events leading to the restoration of GH/IGF system activity after starvation is largely unknown. We, therefore, measured the plasma levels of GH, IGF-I and IGF-II and the expression of the GH/IGF system in liver and muscle. Starvation increased the plasma GH level and 1 day of refeeding completely restored it (1.10 +/- 0.27 vs 1.12 +/- 0.28 ng/ml). Thereafter, plasma GH continued to decrease until day 7 and returned to control values from day 15. Starvation decreased plasma IGF-I and IGF-II and refeeding raised plasma IGF-I only from day 4. In contrast, the plasma IGF-II level doubled after 1 day's refeeding (26.5 +/- 1.9 vs 44.0 +/- 3.4 ng/ml; P < 0.01). Starved fish exhibited higher GH receptor (GHR)1 mRNA abundance in liver and muscle than in controls, whereas GHR2 mRNA abundance was increased only in muscle. In liver, 1 day of refeeding, decreased GHR1 (twofold), but increased GHR2 mRNA abundance (twofold). Thereafter, a progressive return to normal values was observed. Liver IGFBP-4 mRNA abundance was lowered in starved fish followed by a progressive restoration during refeeding. Starvation had no effect on liver IGFBP-2 and IGFBP-6 mRNA abundance, whereas refeeding provoked a peak of IGFBP-2 and IGFBP-6 expression at day 7. In muscle, starvation led to a decrease of the IGFBP-2 mRNA level, which was restored only from day 7. IGFBP-4 mRNA abundance in starved fish was lower than in the controls and refeeding led to a transient upregulation (sevenfold) of IGFBP-4 gene at day 1. IGF-I, IGFBP-5, and IGFBP-related protein 1 (rP1) expression profiles were similar, showing a decrease of expression after starvation, a first peak of expression at day 2, a second peak at day 7, and a return to normal value from day 15. Moreover, IGF-I, IGFBP-5, and IGFBP-rP1 mRNA abundance were positively correlated (r = 0.6-0.8; P < 0.0001). In conclusion, plasma IGF-I was restored later than plasma GH level, which suggests that plasma IGF-I levels cannot account for plasma GH changes. The coordinated regulation of IGF-I, IGFBP-5, and IGFBP-rP1 expression would be a signature for the resumption of myogenic activity.

Effects of fasting therapy on irritable bowel syndrome

How to treat patients with irritable bowel syndrome (IBS) who do not respond to pharmacotherapy is an unsolved problem. Psychotherapy, which has been reported on in previous studies, is available only in specific centers. We describe in this study a novel and simple psychotherapy; that is, the fasting therapy (FT) for treatment of patients with IBS. Of 84 inpatients with IBS, 58 patients who still had moderate to severe IBS symptoms after 4-week basic treatment were investigated retrospectively. Of the 58 patients enrolled in this study, 36 underwent FT, whereas the remaining 22 received a consecutive basic treatment (control therapy). There were no significant differences in the 4-point severity scales of gastrointestinal and psychological symptoms between the 2 groups before the start of FT. The basic treatment consisted of pharmacotherapy and brief psychotherapy, whereas the FT consisted of 10 days of starvation followed by 5 days of refeeding. Changes in scores of symptoms before and after each treatment were analyzed. FT significantly improved 7 out of the 10 symptoms assessed; that is, abdominal pain-discomfort (p < .001), abdominal distension (p < .001), diarrhea (p < .001), anorexia (p = .02), nausea (p < .01), anxiety (p < .001), and interference with life in general (p < .001). However, the control therapy significantly improved only 3 out of the 10 symptoms assessed; that is, abdominal pain-discomfort (p = .03), abdominal distension (p < .01), and interference with life (p = .01). Our results suggest that FT may have beneficial effects on intractable patients with IBS.

Expression of purinergic receptors in the hypothalamus of the rat is modified by reduced food availability

ATP-sensitive P2 receptors are suggested to play an important role in the cerebral signal transduction. We examined the expression of the P2Y1 receptor and the possibly downstream-related neuronal nitric oxide synthase (nNOS) in the hypothalamus of rats food-restricted for 3 or 10 days and rats refed after a restriction of 10 days. The restriction caused a reduction of the body weight and plasma triacylglyceride, an increase of non-esterified fatty acid levels correlating with a decrease of leptin levels and an enhancement of plasma corticosterone. All changes returned to basal levels after refeeding. The restriction induced an enhanced intake within 30 min after food presentation and a reduction in the latency. Interestingly, the latter was not abolished by refeeding. The daily food intake induced by refeeding was enhanced at the first day only. The expression of hypothalamic P2Y1 receptor/nNOS mRNA and protein and of leptin receptor mRNA were enhanced after restricted feeding. These changes were abolished after 3 days of refeeding. Immunofluorescence studies indicated that P2Y1 receptor and nNOS immunoreactivities are present in the dorsomedial, ventromedial and lateral hypothalamus and in the nucleus arcuatus. P2Y1 receptor-positive cells were partially also nNOS-positive. The P2Y1 receptor labeling was restricted to cell bodies of obviously non-glial cells, whereas nNOS labeling could be detected also at cellular processes of these cells. In the nucleus arcuatus, astrocytes were identified, expressing P2Y1 receptors at cell bodies and cellular processes. The data suggest that restricted feeding may enhance the sensitivity of the hypothalamus to extracellular ADP/ATP by regulation of the expression of P2Y1 receptors and possibly of their signal transduction pathway via nitric oxide production.

Fasting therapy for functional gastrointestinal disorders

The fasting therapy (FT) was developed as novel and simple psychotherapy for psychosomatic diseases from our laboratory. FT consisted of 10 days of starvation with 5 days of re-feeding. FT requires an individual room in the health care utilization due to social isolation. Although FT has empirically been used in Japan, little has known about precise mechanisms on the efficacy of this treatment. However, there are some interesting findings to suggest that this treatment may be effective for functional gastrointestinal disorders. It was reported that FT brought not only reduction of physical symptoms but also improvement of psychological abnormalities in intractable patients with irritable bowel syndrome (IBS). Further studies should be needed to clarify this treatment can be one of powerful psychological treatments for functional gastrointestinal disorders and the other psychosomatic diseases.

Early induction of TGF‐β1 through a fasting–re‐feeding regimen promotes liver carcinogenesis by a sub‐initiating dose of diethylnitrosamine

We previously reported that a sub-necrogenic dose (20 mg/kg) of diethylnitrosamine (DENA) can induce the development of liver cancer when rats undergo a fasting-re-feeding regimen. The present study was undertaken to establish whether fasting followed by re-feeding builds up mechanisms able to trigger liver fibrosis, eventually leading to cirrhosis and cancer. Adult male rats, for fasted 4 days, were given 20 mg/kg of DENA after 1 day of re-feeding; in parallel, consistently fed animals receiving 20 mg/kg (sub-necrogenic) or 200 mg/kg (necrogenic dose) of DENA were used as negative and positive controls, respectively. All three groups were then subjected to the 2-acetylaminofluorene/carbon tetrachloride promoting regimen. Fasting induced moderate apoptosis in liver tissue, as evidenced by increased levels of transforming growth factor-beta1 (TGF-beta1) and Bax proteins and by a dramatic drop in the level of Bcl-2. Subsequent re-feeding caused all changes to revert except TGF-beta1 up-regulation. Histological findings of inflammation and fibrosis were consistently associated with increased production of TGF-beta1, the inflammatory cytokine with the most pronounced profibrogenic action. Thus, up-regulation of TGF-beta1 expression appears as a major mechanism by which the fasting-re-feeding regimen predisposes to initiation and promotion of liver carcinogenesis in rats. Avoiding fasting-re-feeding could be considered in the nutritional status of patients with liver fibrosis.

Alterations in expression of genes associated with muscle metabolism and growth during nutritional restriction and refeeding in rainbow trout

Rainbow trout, as well as many other species of fish, demonstrate the ability to survive starvation for long periods of time. During starvation, growth rate is decreased and muscle exhibits signs of wasting. However, upon resumption of feeding, accelerated growth is often observed. Alterations in muscle metabolism occur during feed restriction and refeeding, although the ways in which these alterations affect the molecular pathways that control muscle growth have not been fully determined. To analyze changes in muscle metabolism and growth during starvation and refeeding, real-time PCR was used to test the expression of six metabolic-related genes and eight muscle-specific genes in rainbow trout white muscle prior to and after 30 days of starvation, and after 4 and 14 days of refeeding. The six metabolic-related genes chosen are indicative of specific metabolic pathways: glycolysis, glycogenesis, gluconeogenesis, the pentose phosphate pathway, and fatty acid formation. The eight muscle specific genes chosen are key components in muscle growth and structural integrity, i.e., MRFs, MEFs, myostatins, and myosin. Alterations in expression of the tested metabolic-related genes and muscle-specific genes suggest that during both starvation and refeeding, changes in specific metabolic pathways initiate shifts in muscle that result mainly in the modification of myotube hypertrophy. The expression levels of many of the metabolic-related genes were altered during the refeeding period compared to those observed before the starvation period began. However, the accelerated growth often observed during refeeding is likely driven by changes in normal muscle metabolism, and the altered expression observed here may be a demonstration of those changes.

Homeostatic responses to caloric restriction: influence of background metabolic rate

The biological responses to caloric restriction (CR) are generally examined in rats with elevated metabolic rates due to being housed at ambient temperatures (T(a)) below the zone of thermoneutrality. We determined the physiological and behavioral responses to 2 wk of 30-40% CR in male FBNF1 rats housed in cool (T(a) = 12 degrees C) or thermoneutral (TMN; T(a) = 30 degrees C) conditions. Rats were instrumented with telemetry devices and housed continuously in home-cage calorimeters for the entire experiment. At baseline, rats housed in cool T(a) had reduced rate of weight gain; thus a mild CR (5%) group at thermoneutrality for weight maintenance was also studied. Rats housed in cool T(a) exhibited elevated caloric intake (cool = 77 +/- 1; TMN = 54 +/- 2 kcal), oxygen consumption (Vo(2); cool = 9.9 +/- 0.1; TMN = 5.5 +/- 0.1 ml/min), mean arterial pressure (cool = 103 +/- 1; TMN = 80 +/- 2 mmHg), and heart rate (cool = 374 +/- 3; TMN = 275 +/- 4 beats/min). Cool-CR rats exhibited greater CR-induced weight loss (cool = -62 +/- 3; TMN = -42 +/- 3 g) and reductions in Vo(2) (cool = -2.6 +/- 0.1; TMN = -1.5 +/- 0.1 ml/min) but similar CR-induced reductions in heart rate (cool = -59 +/- 1; TMN= -51 +/- 7 beats/min). CR had no effect on arterial blood pressure or locomotor activity in either group. Unexpectedly, weight maintenance produced significant reductions in Vo(2) and heart rate. At thermoneutrality, a single day of refeeding effectively abolished CR-induced reductions in Vo(2) and heart rate. The results reveal that rats with low or high baseline metabolic rate exhibit comparable compensatory reductions in Vo(2) and heart rate and suggest that T(a) can be used to modulate the metabolic background on which the more prolonged effects of CR can be studied.

Neuropeptide Y content in the hypothalamic paraventricular nucleus responds to fasting and refeeding in broiler chickens

To examine the neural mechanism by which hypothalamic neuropeptide Y (NPY) regulates energy homeostasis and feeding behavior in commercial broilers, we measured NPY content in several hypothalamic regions of birds that were fasted and then refed. After fasting for 48 and 72 h, body weight significantly decreased, and food intake significantly increased during the subsequent refeeding. The lost body weight was not restored to ad libitum feeding levels even after 3 days of refeeding. Plasma glucose concentration and body fat content significantly decreased and plasma non-esterified fatty acid (NEFA) concentration significantly increased after 48- and 72-h fasting. Refeeding for 24 h restored plasma metabolites and body fat content to pre-fasting levels. NPY content in the paraventricular nucleus (PVN) and infundibular nucleus significantly increased during fasting, and NPY content of the PVN was restored to pre-fasting levels after 24-h refeeding. However, there was no significant change in the NPY content of the lateral hypothalamic area during fasting or refeeding. The present results of changes in the hypothalamic NPY content during fasting and refeeding support the hypothesis that NPY plays a central role in regulation of energy homeostasis, with especially important effect on feeding behavior and body weight in broiler chickens.

Energy metabolism and nutrient oxidation in young pigs and rats during feeding, starvation and re-feeding

The investigation included individual measurements of energy metabolism and oxidation of nutrients in 12 castrated male pigs ( Sus scrofa) (20–40 kg) and 12 male rats ( Rattus norvegicus) (65–105 g). Measurements were carried out in 5–6 days balance periods with ad libitum feeding, followed by 3–4 days of starvation and 4 days of re-feeding. O 2 consumption and CO 2 production were measured by open-air-circuit respiration units. In the feeding period, protein retention in relation to metabolic live mass (kg 0.75) was identical for pigs and rats, while there was a tendency of a higher fat retention in pigs than in rats. A substantial part of digested carbohydrate was not oxidized, but transferred to fat metabolism without significant differences ( P>0.05) between pigs and rats (18% vs. 22%). During starvation, nitrogen excretion in urine decreased to 226 mg/kg 0.75 in pigs and to 429 mg/kg 0.75 in rats, indicating a lower rate of body protein degradation in pigs. Heat production was reduced to 592 and 338 kJ/kg 0.75, while the contribution of heat from oxidation of protein (OXP), carbohydrate (OXCHO) and fat (OXF) showed the same pattern for pigs and rats during all periods. Heat production during feeding and re-feeding was covered by OXP+OXCHO with no OXF and reversibly after 2 days of starvation by OXP+OXF with no OXCHO. The rat may be a suitable model for pigs regarding general patterns of quantitative nutrient partition, but any direct application of results measured with rats to pigs shall be taken cautiously, keeping in mind that modern pigs have been selected for a high growth rate and protein deposition which has not been the case for the laboratory rat.

Energy reserves during food deprivation and compensatory growth in juvenile roach: the importance of season and temperature

The effect of 21 days of starvation, followed by a period of compensatory growth during refeeding, was studied in juvenile roach Rutilus rutilus during winter and summer, at 4, 20 and 27° C acclimation temperature and at a constant photoperiod (12L : 12D). Although light conditions were the same during summer and winter experiments and fish were acclimated to the same temperatures, there were significant differences in a range of variables between summer and winter. Generally winter fish were better prepared to face starvation than summer fish, especially when acclimated at a realistic cold season water temperature of 4° C. In winter, the cold acclimated fish had a two to three‐fold larger relative liver size with an approximately double fractional lipid content, in comparison to summer animals at the same temperature. Their white muscle protein and glycogen concentration, but not their lipid content, were significantly higher. Season, independent of photoperiod or reproductive cycle, was therefore an important factor that determined the physiological status of the animal, and should generally be taken into account when fish are acclimated to different temperature regimes. There were no significant differences between seasons with respect to growth. Juvenile roach showed compensatory growth at all three acclimation temperatures with maximal rates of compensatory growth at 27° C. The replenishment of body energy stores, which were utilized during the starvation period, was responsible for the observed mass gain at 4° C. The contribution of the different energy resources (protein, glycogen and lipid) was dependent on acclimation temperature. In 20 and 27° C acclimated roach, the energetic needs during food deprivation were met by metabolizing white muscle energy stores. While the concentration of white muscle glycogen had decreased after the fasting period, the concentrations of white muscle lipid and protein remained more or less constant. The mobilization of protein and fat was revealed by the reduced size of the muscle after fasting, which was reflected in a decrease in condition factor. At 20° C, liver lipids and glycogen were mobilized, which caused a decrease both in the relative liver size and in the concentration of these substrates. Liver size was also decreased after fasting in the 4° C acclimated fish, but the substrate concentrations remained stable. This experimental group additionally utilized white muscle glycogen during food deprivation. Almost all measured variables were back at the control level within 7 days of refeeding.

The effects of fasting and refeeding on biochemical parameters in the red-legged partridge ( Alectoris rufa)

Every year, about three million farm-reared red-legged partridges are released in Spain for hunting purposes, most of them for restocking where the success is very low. Some factors such as the sudden shift from abundant commercial energy-rich diet to natural food and absence of experience at looking for natural food resources could cause a rapid physical deterioration, being more vulnerable to predation and disease. To determine fasting capacity in red-legged partridges, we fasted 32 birds during 4 days and then we refed them, collecting blood samples and body masses during the experiment. Red-legged partridges entered in the third critic phase of fasting, in which body proteins became the main energetic source, after 48 h of fasting. The fasting period caused an average loss of 17% of their initial body mass and only attained a 93.6% of their initial body mass after 24 days of refeeding. The limited capacity to resist fasting in the red-legged partridge could be one of the reasons for the low success in restocking, mainly in those where neither supplementary feeding nor acclimation period is allowed for birds before they are finally released.

Diurnal rhythm in heat production and oxidation of carbohydrate and fat in pigs during feeding, starvation and re-feeding.

Diurnal rhythm in heat production (HE), oxidation of carbohydrate (OXCHO) and fat (OXF) was calculated from daily measurements of gas exchange in 12 pigs [20-40 kg live weight, (LW)] during 6 days of near ad libitum feeding, followed by 4 days of starvation and 4 days of re-feeding. All measurements, divided in five times intervals from 12.00 to 8.00, showed the highest values of HE, reflecting the animals' energy requirements, between 12.00 and 16.00 gradually declining to the lowest values between 4.00 and 8.00. The values measured in the interval 4.00-8.00 were considered as a basal metabolic rate (BMR), being in all measurements 25% lower than during 12.00-4.00. The lowest BMR was measured on the fourth day of starvation (21.7 kJ/h.kg(0.75)). By transition from feeding to starvation, OXCHO declined gradually, but was for 16 h able to cover the energy requirement with no contribution from OXF. The decline in OXCHO proceeded for 40 h and reached zero between 4.00 and 8.00 on the first day of starvation with the energy requirement being covered by OXF. The HE during starvation was 25-30% lower than during feeding caused by absence of feed-induced thermogenesis and by the transition from OXCHO to OXF. Immediately after re-feeding dietary carbohydrates were oxidized, however, there was still a substantial OXF, proceeding until the next feeding. From the second day of re-feeding the contribution of substrates to the total HE was re-established with no OXF and the same level of HE as during feeding.

Energy metabolism and substrate oxidation in pigs during feeding, starvation and re-feeding.

Energy metabolism and substrate oxidation was measured in 12 growing pigs by means of indirect calorimetry and nutrient balances. The measurements were carried out during 5 days of feeding followed by 4 days of starvation and 5 days of re-feeding. During the feeding period, dietary carbohydrates were the main energy source, sufficient to cover energy requirements without oxidation of fat. Starvation reduced the total heat production and affected the oxidation pattern by reducing protein oxidation and shifting from carbohydrate to fat oxidation. On the second day of starvation, the main energy source was body fat and there was no oxidation of carbohydrate. On the second day of re-feeding, the heat production reached the same level as during the feeding period. Also, during the second day of re-feeding, the pattern of nutrient oxidation was similar to the feeding period with oxidation of carbohydrate providing 90% and oxidation of protein providing 10% of the total heat production. There was no fat oxidation. The results demonstrated that the growing pigs were able to re-establish oxidative patterns already 2 days after re-alimentation.

Reduction of anxiety after restricted feeding in the rat: implication for eating disorders

Background Eating-disorder patients exhibit not only abnormal eating attitudes but also pathologic anxiety-like behaviors. The specific nature of the relationship between dieting and anxiety-like behavior is unknown. Methods To investigate the adaptational changes that resulted from chronic restricted scheduled feeding (2-hour access per day for 2 weeks) and subsequent free refeeding, longitudinal changes in the microstructure of feeding behavior were studied in male rats. To study the relationship between restricted feeding and anxiety-like behavior, separate rats were tested in the elevated plus-maze under the following conditions: 1) free feeding; 2) acute food restriction (2-hour access for 1 day); 3) chronic food restriction (for 10 days); or 4) postrecovery (after 10 days of free feeding subsequent to chronic food restriction). Results The effects of chronic food restriction on meal structure diminished within a few days after refeeding. Decreased anxiety-like behavior was seen during acute and chronic food restriction and did not reflect nonspecific behavioral activation. Anxiolytic-like effects persisted after 10 days of refeeding. Conclusions Chronic food restriction produced reductions in anxiety-like behavior that persisted beyond the normalization of food intake patterns. The findings might have etiologic and pathophysiologic relevance for the restrained eating pattern in eating-disorder patients with comorbid anxious symptoms.

Implementing Clinical Pathways for Patients Admitted to a Medical Service: Lessons Learned

In an attempt to improve quality of care for patients admitted to our medical service we have implemented the use of pathways. These are printed standards of care and a mechanism for daily multidisciplinary documentation. The goals of our pathways are to: improve quality using printed standards of care; improve documentation of the care delivered; improve communication about daily goals between all team members, patients and families; standardize our in-patient chart format throughout the hospital; and increase efficiency of care. Pathways were designed to provide physicians and nurses with the standards for care and provide a mechanism for multidisciplinary documentation on our in-patient charts. We now have 2 pathways in use on our medical service. One is a clinical care plan (CCP) and the other is a Pancreatitis Pathway (PP) for patients admitted with acute pancreatitis and the other a guideline for care for all patients. The pathways were developed by teams including attending physicians (General Internists and Gastroenterologists), medicine house officers, nurses, and care coordinators. The pathways are used for all patients admitted to our medical service if they are admitted to one of 2 floors. This paper includes a comparison of outcomes for our first 9 patients who were managed using the pancreatitis pathway versus 7 patients cared for without the pathway. Significant differences in the pancreatitis pathway treated patients included: 1) less intense pain on day 2, (P = 0.04); 2) less pain on day of refeeding (P = 0.004); and 3) less IV fluids administered (P = 0.05). We also describe several lessons we have learned about using pathways for in-patients on a medical service in an academic medical center. We have learned the following lessons. Nursing documentation is improved. Physicians need ongoing encouragement and education about the value of pathways. There is considerable work involved for unit coordinators, care coordinators, and nursing in using pathways on a medical-surgical floor. There must be physician and nurse champions. There must be ongoing feedback to users. There must be input from users and edits. We believe the use of pathways as a process to remind clinicians of quality standards will improve the care of our patients by decreasing variation, improving team communication, and enhancing patient and family education.

Effects of fasting and refeeding on jejunal morphology and cellular activity in rats in relation to depletion of body stores

Background: Intestinal mucosa atrophy following a period of starvation characterized by the mobilization of fat stores for energy expenditure (phase II) worsen after a long fast marked by an increase in protein catabolism (phase III). However, the morphology of the jejunum is completely restored after 3 days of refeeding. The aim of this study was to determine the mechanisms involved in the rapid jejunal restoration following the critical phase III. Methods: Jejunal structure was observed through conventional and environmental scanning electron microscopy, whilst cellular dynamics were studied using classical optic microscopy tools and immunohistochemistry. Results: Mucosal structural atrophy during fasting proved to worsen over the two phases. During phase II, apoptosis is still present at the tip of the villi, the number of mitosis in crypts showed a 30% decrease and a transient drop in cell migration is observed. During phase III, however, an 85% rise in mitosis was noticed along with an increase in cell migration and the disappearance of apoptotic cells at the villus tips. This increased cell renewal continues after food ingestion. Conclusions: Starved rats appeared to be in a phase of energy sparing in phase II, with depressed cellular events in the intestinal mucosa. In phase III, however, the preservation of functional cells and the early increase in crypt cell proliferation should prepare the mucosa to refeeding and could explain why jejunal repairs are complete after 3 days of refeeding following either phase II or phase III.

Enzyme-Linked Immunosorbent Assay for Bovine Apolipoprotein A-IV

The present report describes an enzyme-linked immunosorbent assay for bovine apolipoprotein (apo) A-IV. This assay was applied to the determination of its concentration and distribution in sera from cattle. The distribution of apoA-IV in lipoprotein fractions separated by ultracentrifugation was mostly recovered in the non-lipoprotein fractions (d>1.21 g/ml, 90%), but, in the case of gel filtration chromatography, apoA-IV was mainly eluted in HDL and non-lipoprotein fractions. The apoA-IV concentrations during early, mid- and late lactating stages in cows were significantly higher than during the nonlactating stage (p<0.05). From early to late lactating stages, the concentration of apoA-IV was unaltered. After 4 days of fasting, the concentration of plasma apoA-IV had decreased significantly (p<0.05) at days 3 and 4, and was returned to the basal level by 3 days of refeeding. These results suggested that the concentration of apoA-IV is modified by nutritional conditions.

Effects of fasting on circulating IGF-binding proteins, glucose, and cortisol in channel catfish ( Ictalurus punctatus)

The effects of fasting on IGF-binding proteins (IGFBPs), glucose, and cortisol in channel catfish were examined. Fed fish (controls) were compared to 14-, 30-, and 45-day fasted fish and 45-day fasted fish refed for 15 additional days. Body length and weight changes, condition factor (CF), hepatosomatic index (HSI), and plasma glucose and cortisol were assessed to determine growth and metabolic status. Body length and growth rates were inhibited ( P<0.05) after 14, 30, and 45 days of fasting. The 14-, 30-, and 45-day fasted fish exhibited hypoglycemia and reduced CF and HSI. Cortisol levels were increased (22.8±15.2 ng/ml versus 4.7±3.9 ng/ml) in 30-day fasted fish compared to fed controls ( P<0.05). Associated with the increase in cortisol in fasted fish was a concomitant increase in plasma levels of a 20-kDa IGFBP through day 45. A 35- and a 45-kDa IGFBP were also identified but were similar between fed and unfed fish throughout the experiment. At the end of 15 days of refeeding, 20-kDa IGFBP, glucose, and cortisol levels were similar to fed controls. Refeeding also caused an increase in growth rates. These results suggest the existence of a catfish counter part to mammalian IGFBP-1, similar to lower molecular mass IGFBPs reported in other species of fish. These results also suggest that a 20-kDa IGFBP is upregulated during fasting-induced growth inhibition of channel catfish and provide additional evidence of the conserved nature of the IGF-IGFBP-growth axis in fish.

Attenuation by leptin of the effects of fasting on ovarian function in hens (Gallus domesticus)

Thirty-four-week-old laying hens received injections of recombinant chicken leptin to assess the role of leptin in avian ovarian function. In the first experiment, the hens (n=60) were divided into three groups: (i). fed ad libitum; (ii). fasted; and (iii). fasted + leptin. Hens were fasted for 5 days and those treated with leptin received 250 microg leptin kg-1 body weight twice a day, i.p. In the second experiment, the hens (n=72) were divided into four groups: (i). fed ad libitum; (ii). fasted; (iii). fasted + leptin given only during fasting (5 days); or (iv). fasted and leptin given during both fasting and 5 days of re-feeding (10 days). LH was measured in blood plasma, and progesterone and oestradiol were measured in blood plasma and the ovary by radioimmunoassay. Apoptosis was examined in the walls of the three largest yellow hierarchical follicles (F3-F1; F3 25-35 mm) by the TdT-mediated dUTP nick-end labelling method. Results showed that the injections of leptin during fasting: (i). delayed cessation of egg laying; (ii). attenuated regression of yellow hierarchical follicles; (iii). altered ovarian steroidogenesis; and (iv). abolished the fasting-induced apoptosis in the wall of F3-F1 follicles during the first 2 days of fasting and partially attenuated apoptosis after 5 days of fasting. Prolongation of leptin injections into the re-feeding period considerably delayed the restoration of the ovary. Expression of leptin receptor in laying hens was determined by RT-PCR. The highest expression of leptin receptor was observed in the hypothalamus. Lower receptor mRNA expression was found in the hypophysis, whereas the lowest expression was observed in the ovary. Within the ovary, a relatively high expression of leptin receptor was found in the stroma with cortical follicles <1 mm, the wall of white (1-8 mm) and small yellow follicles (>8-12 mm), and the granulosa layer of F3 follicles. The expression of leptin receptor in the granulosa layer of F2 and F1 follicles was barely detectable. This was in contrast to a much higher expression of leptin receptor maintained in the theca layer of F3-F1 follicles. The present results indicate that in chickens leptin might be involved in the adaptation to starvation due to attenuation of follicular apoptosis. The presence of leptin receptors in the ovary indicates the possibility of a peripheral effect of the hormone.