Showing papers on "Low protein published in 1999"

A low protein diet alters the balance of islet cell replication and apoptosis in the fetal and neonatal rat and is associated with a reduced pancreatic expression of insulin-like growth factor-II

Abstract: A programmed turnover of pancreatic beta cells occurs in the neonatal rat involving a loss of beta cells by apoptosis, and their replacement by islet cell replication and neogenesis. The timing of apoptosis is associated with a loss of expression of a survival factor, insulin-like growth factor-II (IGF-II), in the pancreatic islets. Offspring from rats chronically fed a low protein isocalorific diet (LP) exhibit a reduced pancreatic beta cell mass at birth and a reduced insulin secretion in later life. This study therefore investigated the impact of LP on islet cell ontogeny in the late fetal and neonatal rat, and any associated changes in the presence of IGFs and their binding proteins (IGFBPs). Pregnant Wistar rats were fed either LP (8% protein) or normal (C) (20% protein) chow from shortly after conception until the offspring were 21 days postnatal (PN). Bromo-deoxyuridine (BrdU) was administered 1 h before rats were killed and pancreata removed from animals between 19.5 days fetal life and postnatal day 21. Offspring of rats given LP diet had reduced birthweight, pancreatic beta cell mass, and pancreas insulin content, with smaller islets compared with control fed animals, which persisted to weaning. Histological analysis showed that islets from pups given LP diet had a lower nuclear labeling index with BrdU in the beta cells, although, paradoxically, more beta cells showed immunoreactivity for proliferating cell nuclear antigen (PCNA). Because PCNA is present in G1 as well as S phase of the cell cycle, we quantified the number of beta cells immunopositive for cyclin D1, a marker of G1, and NEK2, an indicator of cells in G2 and mitosis, More beta cells in islets from LP-fed animals contained cyclin D1, but less contained NEK2 than did those in controls. This suggests that the beta cell cycle may have a prolonged G1 phase in LP-fed animals in vivo. Offspring of rats given C diet had a low rate of islet cell apoptosis detected by the TUNEL method in fetal and neonatal life (1-2%), with a transient increase to 8% at PN day 14. Offspring of rats receiving LP diet demonstrated a significantly greater level of islet cell apoptosis at every age, rising to 15% at PN 14. IGF-II mRNA was quantified in whole pancreas and was significantly reduced in LP-fed animals at ages up to PN day 10. IGF-II immunoreactivity within the islets of LP-fed rats was also less apparent, but no changes were seen in immunoreactive IGF-I or IGFBPs-2 to -5. These findings show that LP diet changes the balance of beta cell replication and apoptosis in fetal and neonatal neonatal life, which may involve an altered length of beta cell cycle, and contribute to the smaller islet size and impaired insulin release seen in later life. A reduced pancreatic expression of IGF-II may contribute to the lower beta cell proliferation rate and increased apoptosis seen in the fetus and neonate after feeding LP diet.

Pharmacokinetics and pharmacodynamics of the nitroimidazole antimicrobials.

Abstract: Metronidazole, the prototype nitroimidazole antimicrobial, was originally introduced to treat Trichomonas vaginalis, but is now used for the treatment of anaerobic and protozoal infections. The nitroimidazoles are bactericidal through toxic metabolites which cause DNA strand breakage. Resistance, both clinical and microbiological, has been described only rarely. Metronidazole given orally is absorbed almost completely, with bioavailability > 90% for tablets; absorption is unaffected by infection. Rectal and intravaginal absorption are 67 to 82%, and 20 to 56%, of the dose, respectively. Metronidazole is distributed widely and has low protein binding (< 20%). The volume of distribution at steady state in adults is 0.51 to 1.1 L/kg. Metronidazole reaches 60 to 100% of plasma concentrations in most tissues studied, including the central nervous system, but does not reach high concentrations in placental tissue. Metronidazole is extensively metabolised by the liver to 5 metabolites. The hydroxy metabolite has biological activity of 30 to 65% and a longer elimination half-life than the parent compound. The majority of metronidazole and its metabolites are excreted in urine and faeces, with less than 12% excreted unchanged in urine. The pharmacokinetics of metronidazole are unaffected by acute or chronic renal failure, haemodialysis, continuous ambulatory peritoneal dialysis, age, pregnancy or enteric disease. Renal dysfunction reduces the elimination of metronidazole metabolites; however, no toxicity has been documented and dosage alterations are unnecessary. Liver disease leads to a decreased clearance of metronidazole and dosage reduction is recommended. Recent pharmacodynamic studies of metronidazole have demonstrated activity for 12 to 24 hours after administration of metronidazole 1 g. The post-antibiotic effect of metronidazole extends beyond 3 hours after the concentration falls below the minimum inhibitory concentration (MIC). The concentration-dependent bactericidal activity, prolonged half-life and sustained activity in plasma support the clinical evaluation of higher doses of metronidazole given less frequently. Metronidazole-containing regimens for Helicobacter pylori in combination with proton pump inhibitors demonstrate higher success rates than antimicrobial regimens alone. The pharmacokinetics of metronidazole in gastric fluid appear contradictory to these results, since omeprazole reduces peak drug concentration and area under the concentration-time curve for metronidazole and its hydroxy metabolite; however, concentrations remain above the MIC. Other members of this class include tinidazole, ornidazole and secnidazole. They are also well absorbed and distributed after oral administration. Their only distinguishing features are prolonged half-lives compared with metronidazole. The choice of nitroimidazole may be influenced by the longer administration intervals possible with other members of this class; however, metronidazole remains the predominant antimicrobial for anaerobic and protozoal infections.

Dynamics of the nuclear lamina as monitored by GFP-tagged A-type lamins

Abstract: The behavior of chimeric proteins consisting of A-type lamins and green fluorescent protein (GFP) was studied to investigate the localization and dynamics of nuclear lamins in living cells. Cell line CHO-K1 was transfected with cDNA constructs encoding fusion proteins of lamin A-GFP, lamin Adelta10-GFP, or lamin C-GFP. In the interphase nucleus lamin-GFP fluorescence showed a perinuclear localization and incorporation into the lamina for all three constructs. Our findings show for the first time that the newly discovered lamin A 10 protein is localized to the nuclear membrane. The GFP-tagged lamins were processed and behaved similarly to the endogenous lamin molecules, at least in cells that expressed physiological levels of the GFP-lamins. In addition to the typical perinuclear localization, in the majority of transfected cells each individual A-type lamin-GFP revealed an extensive collection of branching intra- and trans-nuclear tubular structures, which showed a clear preference for a vertical orientation. Time-lapse studies of 3-D reconstructed interphase cells showed a remarkable stability in both number and location of these structures over time, while the lamina showed considerable dynamic movements, consisting of folding and indentation of large parts of the lamina. Fluorescence recovery after bleaching studies revealed a low protein turnover of both tubular and lamina-associated lamins. Repetitive bleaching of intranuclear areas revealed the presence of an insoluble intranuclear fraction of A-type lamins. Time-lapse studies of mitotic cells showed that reformation of the lamina and the tubular structures consisting of A-type lamins did not occur until after cytokinesis was completed.

Changes in renal function during weight loss induced by high vs low-protein low-fat diets in overweight subjects.

Abstract: Changes in renal function during weight loss induced by high vs low-protein low-fat diets in overweight subjects

Changes in Bone Turnover in Young Women Consuming Different Levels of Dietary Protein

Abstract: Although high protein diets are known to increase urinary calcium excretion and induce negative calcium balance, the impact of dietary protein on bone turnover and fractures is controversial. We therefore evaluated the effect of dietary protein on markers of bone turnover in 16 healthy young women. The experiment consisted of 2 weeks of a well balanced diet containing moderate amounts of calcium, sodium, and protein followed by 4 days of an experimental diet containing one of three levels of protein (low, medium, or high). On day 4, serum and urinary calcium, serum PTH, 1,25-dihydroxyvitamin D, serum osteocalcin, bone-specific alkaline phosphatase, and urinary N-telopeptide excretion were measured. Urinary calcium excretion was significantly higher on the high than on the low protein diet. Secondary hyperparathyroidism occurred on the low protein diet. Urinary N-telopeptide excretion was significantly greater during the high protein than during the low protein intake (48.2 +/- 7.2 vs. 32.7 +/- 5.3 nM bone collagen equivalents/mM creatinine; P < 0.05). There was no increase in osteocalcin or bone-specific alkaline phosphatase when comparing the low to the high diet, suggesting that bone resorption was increased without a compensatory increase in bone formation. Our data suggest that at high levels of dietary protein, at least a portion of the increase in urinary calcium reflects increased bone resorption.

Seasonal and spatial changes in the sediment organic matter of a semi-enclosed marine system (W-Mediterranean Sea)

Abstract: The composition of sediment organic matter and the related role of microphytobenthic biomass have been investigated during one-year in a semi-enclosed marine system (Marsala lagoon, Mediterranean Sea). Monthly samples from June 1993 to May 1994 were analysed for carbohydrate, protein, lipid, photosynthetic pigment and total organic matter. The three main biochemical classes of organic compounds (i.e. carbohydrates, proteins and lipids) showed higher concentrations than reported in the literature. However, photosynthetic pigment was quite low, compared to other enclosed marine basins or Mediterranean coastal lagoons. As a result, the contribution of primary organic carbon to the sedimentary biopolymeric fraction of organic matter was low (on average 2.2%), indicating that most of the sedimentary organic matter in the study area originated from sources other than micro-algae. The sedimentary organic matter, dominated by carbohydrates (on average 51.2%) followed by proteins (39.0%) and lipids (9.8%), as well as the low protein to carbohydrate ratio, indicate the presence of large amounts of non-living and/or aged organic matter. Comparing data on spatial distribution of sedimentary and suspended organic matter, the dynamic balance of resuspension vs. sedimentation along a north-south axis is invoked as one major factor affecting the distribution and composition of the main classes of organic compounds. The macroalgal and vascular plant coverage is suggested to be another major factor affecting both amounts and composition of sedimentary organic matter. The northern area, characterised by partially unvegetated sediments, showed higher amounts of proteins, whilst moving southward and approaching a luxuriant Posidonia oceanica reef, carbohydrates became more important relative to proteins. As only the biopolymeric fraction of sediment organic matter showed significant seasonal changes, the quantity of sediment OM behaves as an emerging property. By contrast, OM quality is strictly connected to algal coverage as well as to episodic inputs of primary organic matter from deposited phytoplankton and/or microphytobenthos. The uncoupling between large amounts and relatively low nutritional value of sedimentary OM suggests that this particular environment behaves as a detrital ‘trap’.

Ketogenic diet reduces spontaneous seizures and mossy fiber sprouting in the kainic acid model.

Abstract: The high fat, low carbohydrate, low protein ketogenic diet (KD) has been used to control refractory epilepsy in children since 1920, although its mechanism of action is unknown. Previous animal studies have shown that the KD can increase acute seizure threshold, but the effect of the KD on the process of epileptogenesis has not been studied. We tested the effect of an experimental KD on epileptogenesis in adult rats using the kainic acid (KA) model. P54 rats underwent KA-induced status epilepticus, followed by assignment to a control diet or a KD consisting of (by weight), 14% protein, 70% fat and no carbohydrate. KD-fed animals tolerated the diet and maintained ketosis. KD-fed rats had significantly fewer and briefer spontaneous recurrent seizures and less supragranular mossy fiber sprouting, although the degree of hippocampal pyramidal cell damage was similar in both groups. These results provide the first evidence that the KD retards epileptogenesis in an experimental model.

A Low-Protein Isocaloric Diet During Gestation Affects Brain Development and Alters Permanently Cerebral Cortex Blood Vessels in Rat Offspring

Abstract: In humans, low birth weight is associated with nonfatal stroke, cardiovascular disease and diabetes at adulthood. The aim of this study was to investigate in rats the effect of early protein restriction, inducing low birth weight, on brain and endocrine pancreas vascularization at birth and to study if such alterations lasted until adulthood. Pregnant rats were fed either 20 or 8% protein isocaloric diets. Control newborns were nursed by their dams fed the 20% protein diet and low protein (LP) pups by dams fed either the 8 or 20% protein diet. The diets given during lactation were maintained until adulthood. The blood vessel density of cerebral cortex analyzed by morphometry in 3-d-old pups from dams fed the 8% protein diet was lower than in control (C). It remained lower at adulthood whether a LP or a C diet was given postnatally. Reduction of vascularization at adulthood induced by the LP diet limited to fetal life seems characteristic for the brain since vascularization of islets of Langerhans was reduced in neonates but normalized at adulthood by a C diet postnatally. Body and brain weights were lower in LP pups and adults. DNA concentration was lower in forebrain and higher in cerebellum in LP pups. In brain of LP adults, DNA, protein, cholesterol and phospholipid concentrations were lower and were restored at adulthood by a normal diet after birth. In conclusion, cerebral cortex of offspring exposed to a LP isocaloric diet during fetal development showed reduced vascularization which remained throughout life.

Ultrasonic atomization for spray drying: a versatile technique for the preparation of protein loaded biodegradable microspheres.

Abstract: Bovine serum albumin (BDA) loaded microspheres with a spherical shape and smooth surface structure were successfully prepared from poly(lactide-co-glycolide) using an ultrasonic nozzle installed in a Niro laboratory spray dryer. Process and formulation parameters were investigated with respect to their influence on microsphere characteristics, such as particle size, loading capacity, and release properties. Preparation of microspheres in yields of more than 50% was achieved using an ultrasonic atomizer connected to a stream of carrier air. Microsphere characteristics could be modified by changing several technological parameters. An increased polymer concentration of the feed generated larger particles with a significantly reduced initial release of the protein. Moreover, microspheres with a smooth surface structure were obtained from the organic polymer solution with the highest viscosity. Microparticles with a low BSA loading showed a large central cavity surrounded by a thin polymer layer in scanning electron microspheres. A high protein loading led to an enlargement of the shell layer, or even to dense particles without any cavities. A continuous in vitro release pattern of BSA was obtained from the particles with low protein loading. Glass transition temperatures (Tg) of the microspheres before and after lyophilization did not differ from those of the BSA loaded particles prepared by spray drying with a rotary atomizer. Analysis of the polymer by gel permeation chromatography indicated that ultrasonication had no effect on polymer molecular weight. Molecular weight and polydispersity of the pure polymer, placebo microspheres prepared by spray drying, and placebo microspheres prepared using the ultrasonic nozzle were in the same range. In conclusion, ultrasonic atomization represents a versatile and reliable technique for the production of protein loaded biodegradable microspheres without inducing a degradation of the polymer matrix. Particle characteristics can be modified by adjusting formulation parameters and atomization conditions in a simple manner.

RFLP markers associated with soybean cyst nematode resistance and seed composition in a ‘Peking’בEssex’ population

Abstract: Soybean cyst nematode (SCN), Heterodera glycines Ichinohe, causes severe damage to soybean [Glycine max (L.) Merr] throughout North America and worldwide. Molecular markers associated with loci conferring SCN resistance would be useful in breeding programs using marker-assisted selection (MAS). In this study, 200 F2:3 families derived from two contrasting parents, SCN-resistant ‘Peking’ with relatively low protein and oil concentrations, and SCN-susceptible ‘Essex’ with high protein and oil concentrations, were used to determine loci underlying the SCN resistance and seed composition. Three different SCN Race isolates (1, 3, and 5) were used to screen both parents and F2:3 families. The parents were surveyed with 216 restriction fragment length polymorphism (RFLP) probes with five different restriction enzymes. Fifty-six were polymorphic and contrasted with trait data from bioassays to identify molecular markers associated with loci controlling resistance to SCN and seed composition. Five RFLP markers, A593 and T005 on linkage group (LG) B, A018 on LG E, and K014 and B072 on LG H, were significantly linked to resistance loci for Race 1 isolate, which jointly explained 57.7% of the total phenotypic variation. Three markers (B072 and K014, both on LG H; T005 on LG B) were associated with resistance to the Race 3 isolate and jointly explained 21.4% of the total phenotypic variation. Two markers (K011 on LG I, A963 on LG E) associated with resistance to the Race 5 isolate together explained 14.0% of the total phenotypic variation. In the same population we also identified two RFLP markers (B072 on LG H, B148 on LG F) associated with loci conferring protein concentration, which jointly explained 32.3% of the total phenotypic variation. Marker B072 was also linked to loci controlling the concentration of seed oil, which explained 21% of the total phenotypic variation. Clustering among quantitative trait loci (QTLs) conditioning resistance to different SCN Race isolates and seed protein and oil concentrations may exist in this population. We believe that markers located near these QTLs could be used to select for new SCN resistance and higher levels of seed protein and oil concentrations in breeding improved soybean cultivars.

Protein malnutrition affects the growth trajectories of the craniofacial skeleton in rats.

Abstract: To investigate the effects of protein malnutrition on a normal growth trajectory, we radiographed Rattus norvegicus from 22 d (weaning) and continuing past adult size. We took measurements from longitudinal radiographs of rats fed a control diet and littermates fed an isocaloric low protein experimental diet. A Gompertz model was fit to each individual rat for body weight and 22 measurements of the craniofacial skeleton, producing parameters that described the rate and timing of growth. We tested for differences in these parameters due to diet, sex and litter with a mixed-model three-way ANOVA. Allometric analysis examined the scaling relationships between and within various regions of the skull. For most measurements, final sizes predicted by the model were not significantly different between rats fed the two diets, although the differences in final measurements showed small, but significant differences in growth between rats in the two diet groups. The instantaneous initial rate of growth, maximum rate of growth and deceleration of growth were significantly higher in the control rats for every measurement. Rats fed the low protein diet grew for a significantly longer period of time. The shape of the neurocranium was relatively conserved between diet groups; however, rats fed the low protein diet had shorter and relatively wider skulls than the controls. These results suggest that functional demands of the viscerocranium were greater after birth, and that growth in this area was faster. The viscerocranium reached functional adult proportions earlier and was therefore more susceptible to epigenetic perturbations such as dietary protein level. Protein malnutrition did not affect many aspects of adult size, but strongly altered the growth trajectory to achieve that size.

Comparison of the supplemental effects of Saccharomyces cerevisiae and antibiotics in low‐protein and high‐fibre diets fed to broiler chickens

Abstract: Two experiments were completed to compare the supplemental effects of yeast Saccharomyces cerevisiae and sub‐therapeutic antibiotics in high‐fibre and low‐protein diets for broiler chicks. In experiment I, yeast was added at 1.5, 3.0 and 6.0gkg‐1 while penicillin, tylosin or neoterramycin were added at 150 mg kg‐1 into different batches of a high‐fibre diet containing 250 g kg‐1 palm kernel meal. Saccharomyces cerevisiae and antibiotics increased BW, BWG, FCR, feed intake, carcass and breast weights above the control. Small intestine weight was reduced by antibiotics, while yeast caused a lower deposition of abdominal fat. Liver weights of different treated broilers were similar. The performance indices were superior on penicillin and 3.0 g kg‐1 yeast compared to other supplements evaluated. In the second experiment, 1.5 and 3.0 g kg‐1 of yeast, and 0.75 mg kg‐1 of procaine penicillin, zinc bacitracin and tylosin were separately added to a 180 g kg‐1 crude protein diet. Performance of the chicks with addi...

In vitro immunoglobulin light chain fibrillogenesis.

Abstract: These techniques permit the production of bulk quantities of fibrils and provide methods for monitoring the kinetics of fibrillogenesis. Experiments performed in the fluorimeter require low protein concentrations, sampling is not necessary (with ThT in situ), and the measured fluorescence signal is indicative of fibril content and is not complicated by the presence of amorphous aggregates. However, ASF using the orbital shaker is a simple, rapid, initial procedure, adequate for screening for fibrillogenic potential, in which multiple experiments can be performed simultaneously and over long periods of incubation. These methods may be used to investigate the fibrillogenesis of VL proteins and BJps as a means of predicting pathogenicity, as well as providing information on the basic biophysical principles underlying light chain aggregation.

Nucleotide-dependent oligomerization of ClpB from Escherichia coli.

Abstract: Self-association of ClpB (a mixture of 95- and 80-kDa subunits) has been studied with gel filtration chromatography, analytical ultracentrifugation, and electron microscopy. Monomeric ClpB predominates at low protein concentration (0.07 mg/mL), while an oligomeric form is highly populated at >4 mg/mL. The oligomer formation is enhanced in the presence of 2 mM ATP or adenosine 5'-O-thiotriphosphate (ATPgammaS). In contrast, 2 mM ADP inhibits full oligomerization of ClpB. The apparent size of the ATP- or ATPgammaS-induced oligomer, as determined by gel filtration, sedimentation velocity and electron microscopy image averaging, and the molecular weight, as determined by sedimentation equilibrium, are consistent with those of a ClpB hexamer. These results indicate that the oligomerization reactions of ClpB are similar to those of other Hsp100 proteins.

Epidemiology and Drug Treatment of Epilepsy in Elderly People

Abstract: Seizures are extremely common in the elderly, with an annual incidence reaching 100 per 100000 people aged over 60 years. Most are precipitated by acute symptomatic illnesses such as stroke or systemic disease. Chronic neurological diseases such as Alzheimer's disease may also cause seizures. The aetiology of seizures in many patients is unknown. Seizures may be situational and subside quickly, but the prevalence of chronic seizures--epilepsy--is as high as 1% in the elderly. The majority of seizures are of partial onset, especially complex partial. Complex partial seizures at this age may be very subtle and hard to diagnose. Generalised-onset seizures also occur, perhaps as a result of diffuse changes with aging or degenerative disease or to a combination of genetic and environmental factors. The prognosis for complete seizure control in this population is relatively favourable. Physiological and disease-related changes with aging result in complex pharmacokinetics. Most changes lead to a need for gentler drug treatment with cautious initiation of drugs at lower dosages. Consideration must be given to renal and hepatic function, protein binding and drug interactions. Determinations of free (unbound) drug concentrations are helpful for highly protein bound drugs. The dosages of newer drugs excreted renally must be adjusted based on creatinine clearance. The dosage of most drugs is determined empirically by careful observation of seizure control and adverse effects. Carbamazepine, valproic acid (sodium valproate), gabapentin and lamotrigine have certain theoretical advantages, but comparative trials of anticonvulsants in the elderly are needed. The ideal drug for older patients would be effective, without neurological toxicity, with low protein binding, a nonparticipant in drug interactions and amenable to once daily administration.

Adsorption of protein layers at the water/air interface as studied by axisymmetric drop and bubble shape analysis.

Abstract: Experimental studies of dynamic and equilibrium surface tensions of aqueous β-casein solutions were performed by using the pendent drop and bubble techniques. A comparison of the results shows good agreement at higher protein concentrations. However, at low protein concentrations a significant difference between the data is observed, caused by the loss of protein molecules from the bulk at the drop surface due to the limited solution reservoir. In bubble experiments the reservoir is larger by more than 2 orders of magnitude so that losses become essential at much lower bulk concentrations. Loss of protein and the diffusional transport in the limited reservoir of a small drop lead to the formation of an adsorption layer with properties quite different from those obtained at a bubble surface.

Dietary soy protein effects on inherited polycystic kidney disease are influenced by gender and protein level.

Abstract: The effects of dietary soy protein compared to casein were examined in male and female CD1-pcy/pcy (pcy) mice with polycystic kidney disease. Animals 10 wk of age were fed purified diets containing either soy protein isolate or casein given at a level of 17.4 or 6% protein. After 13 wk on the diets, body weights and serum concentrations of albumin and protein indicated that protein nutrition was adequate on all diets. Overall, animals fed soy protein versus casein had 28% lower (P = 0.0037) relative kidney weights (g/100 g body wt), 37% lower (P = 0.0089) cyst scores (% cyst area x relative kidney weight), and 25% less (P = 0.0144) kidney water (g). Dietary protein reduction resulted in 30% lower (P = 0.0010) relative kidney weights, 25% lower (P = 0.0327) cyst scores, and 35% less (P = 0.0001) kidney water. Analysis of interactions between main effects revealed that the effects of soy protein on kidney size were significant only in females, and that effects of soy protein on cyst score were significant only in animals on the low protein diets. In addition, differences in kidney weights and cyst score due to protein reduction were significant in animals fed soy protein, but not in those fed casein as the protein source. These results show that both dietary protein source and level significantly affect polycystic kidney disease in pcy animals, with the effects of dietary soy protein being most pronounced in female animals fed the low protein diets and the effects of protein reduction being most pronounced in animals fed soy protein-based diets.

Formation of short-lived protein aggregates directly from the coil in two-state folding.

Abstract: Recent results on the 102 residue protein U1A show that protein aggregation is not always slow and irreversible but may take place transiently in refolding studies on a millisecond time scale. In this study we observe a similar aggregation behavior with the classical two-state protein CI2. Since both U1A and CI2 appear to fold directly from the coil at low protein concentrations, it is likely that the aggregates also form directly from the coil. This is in contrast to the behavior of larger multistate proteins where aggregation occurs in connection to "sticky" intermediates.