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PubMed1993

[Histochemical reaction to NPK fertilizer in the fascicular and reticular zones of the adrenal cortex].

Male, white rats were kept for 30 days in an NPK fertilizer factory. The animals were then sacrificed and the samples of the fascicular and reticular zone of the adrenal gland were analyzed for fat substances. In the fascicular zone, as affected by NPK, the quantity of neutral lipids and triglycerides was decreased, whereas phospholipids were augmented. At the level of reticular zone the lowest rise in the quantity of neutral lipids was found, whereas the amount of the other two sorts of fat substances was conspicuously elevated. Nucleic acids change their presence in that pironynophilia disappeared; in the cells of both zones, the full discharge of RNA took place. Judging by the results obtained, the cells of both fascicular and reticular zone are in the phase of an intensified secretion.

Adrenal Cortex↗
PubMed1997

Optimized gene synthesis, high level expression, isotopic enrichment, and refolding of human interleukin-5.

Structural studies on soluble proteins using nuclear magnetic resonance (NMR) spectroscopy and other structural methods in general require large quantities of isotopically enriched proteins. Human interleukin-5 is a disulfide-linked homodimeric cytokine implicated in asthmatic response. The development of a high yield overexpression system for human interleukin-5 is an important prerequisite to using modern multidimensional NMR in the characterization of the solution structure of the protein and to characterize interactions with a soluble receptor domain. Significant amounts of the protein were expressed using an optimized synthetic gene in a high yield expression system. Gene synthesis was accomplished through the ligation of six oligonucleotides composed of optimized codons. The ligated fragments were further amplified by a polymerase chain reaction and then subcloned into the T7 RNA polymerase based overexpression vector pET11a. However, the induced protein accumulated in the form of inclusion bodies. Initially, the protein was solubilized under denaturing conditions and purified in these denaturing conditions by passage through a single S-200 HR sizing column. Finally, protein refolding was initiated in the presence of 2 M urea followed by dialysis. This protocol yielded 40 mg of biologically active, isotope-enriched protein from 4 liters of minimal medium thus facilitating structural studies by NMR. The strategy described may be of immense value in the production of significant quantities of recombinant, eukaryotic proteins for structural and other studies.

Base Sequence↗
PubMed1992

Genetic expression for type I procollagen in the early stages of flexor tendon healing.

To determine the precise mechanism by which contact tendon healing occurs at the cellular level, the production of pro alpha (I) collagen messenger RNA (mRNA) produced by fibroblasts of healing intrasynovial flexor tendons was determined by an in situ hybridization technique. The repair site and the proximal and distal tendon stumps of repaired tendons treated with early controlled passive mobilization were fixed and buffered in formalin, 3, 7, 10, and 17 days after repair. A complimentary DNA (cDNA) probe corresponding to alpha (I) procollagen mRNA was labeled with [32P]d-CTP. After hybridization, autoradiography, and staining of the sections, the level of procollagen mRNA was assessed by microscopic examination. Rising levels of procollagen mRNA, indicating progressively increasing levels of synthetic collagen activity, were detected in the healing tendons through 10 days. A moderate decrease in procollagen mRNA was seen at 17 days. Genetic expression for procollagen mRNA was localized specifically to the epitenon cells on the tendon surface overlying the repair site and to cells in the gap between the tendon stumps. No detectable expression was noted in endotenon fibroblasts. The finding of high levels of expression for procollagen type I mRNA in the surface layer of healing tendons demonstrates that cells intrinsic to tendon epitenon contribute the greatest quantity of native tendon collagen to the repair site during these important early intervals after tendon suture.

Animals↗
PubMed2000

In vitro IL-12 treatment of peripheral blood mononuclear cells from patients with leukemia or myelodysplastic syndromes: increase in cytotoxicity and reduction in WT1 gene expression.

Interleukin-12 (IL-12) has potent antitumor activities. We examined whether IL-12 enhanced the cytotoxicity of peripheral blood mononuclear cells (PBMNC) and decreased leukemia cells in 30 patients with leukemia or myelodysplastic syndromes (MDS): 12 acute myeloid leukemia (AML) (five in complete remission (CR) and seven in non-CR); six chronic myeloid leukemia (CML); and 12 MDS (three refractory anemia (RA), eight RA with excess of blasts and one chronic myelomonocytic leukemia). PBMNC from patients and five healthy volunteers were cultured at 5 x 10(5)/ml parallel with or without 100 units/ml of IL-12 for 3 days. Cytotoxicity of PBMNC against K562 cells was assessed by flow cytometry. To quantify the amount of leukemia cells, WT1 mRNA was measured by competitive reverse transcription polymerase chain reaction (RT-PCR), since WT1 mRNA is considered as a marker of minimal residual disease (MRD) in leukemia or MDS. The cytotoxicity of non-IL-12-treated PBMNC of 30 patients was 13.4+/-9.3% at the effector to target (E:T) ratio of 20:1, and significantly lower than that of normal subjects (25.7+/-8.4%). The cytotoxicity increased to 30.6+/-17.9% in the IL-12-treated PBMNC. WT1 mRNA in PBMNC of five healthy volunteers was less than 10(3) copies/microg of total RNA. Following the 3-day IL-12 treatment, mean WT1 mRNA of PBMNC was reduced from 10(4.8) to 10(4.2) copies/microg of total RNA in six CML patients, from 10(5.4) to 10(4.8) copies/microg in 12 MDS patients and from 10(5.0) to 10(4.2) copies/microg in five AML patients in CR, but not reduced in five of seven AML in non-CR. These results showed that IL-12 significantly enhanced PBMNC cytotoxicity and decreased the quantity of leukemia cells in PBMNC of most patients with MDS, CML and AML in CR. IL-12 might be of considerable benefit in the elimination of MRD in patients with hematological malignancies.

Adult↗
PubMed1998

Generation of the short RNA transcript in Leishmaniavirus correlates with the growth of its parasite host, Leishmania.

Leishmaniavirus 1 is a double-stranded RNA virus that infects the New World kinetoplastid parasites, Leishmania braziliensis, and Leishmania guyanensis. The isolated virus particles contain an RNA-dependent RNA polymerase which exhibits both transcriptase activity for genome-length plus-strand synthesis and replicase activity for genome-length minus-strand synthesis. Recently, we identified a 320 nucleotide short RNA transcript of Leishmaniavirus 1-4, derived from the 5' end of the viral plus-strand, which is generated by the virus capsid via site-specific cleavage of the full-length positive single-stranded RNA. We have hypothesized that this short RNA transcript functions to regulate the virus life cycle during the growth of its parasite host, Leishmania guyanensis. To address this hypothesis, we measured the relative amount of short RNA transcripts and the absolute number of viral genomes per infected cell from log through stationary phase of the parasite growth cycle. In vitro assays of the viral polymerase showed an overall increase in viral polymerase activity from log growth into stationary phase which mirrored an in vivo increase in the quantity of double-stranded genome as measured by agarose gel electrophoresis. We have developed competitive reverse transcription-polymerase chain reaction (RT-PCR) assays to measure the relative amounts of viral transcripts in infected cells as well as the number of viral genomes per infected cell. The results of these assays show that the amount of full-length virus transcripts peaks in the parasite stationary phase (132 transcripts per cell), and that the short transcript is most abundant in the early stationary phase cells (24 transcripts per cell).

Animals↗
PubMed2004

Substitution Glu480Lys in erythroid band 3 corresponds to the Fr(a) blood group antigen and supports existence of the second ectoplasmic loop of band 3.

BACKGROUND: Polymorphisms in extracellular loops of RBC band 3 correspond to antigens of the Diego blood group system. Of the seven putative extracellular loops, no mutations have until recently been found in the second, fifth, and sixth loops. We detected a substitution Glu480Lys that would be located in its second ectoplasmic loop. We hypothesized the substitution may underlie a novel antigen of the Diego system. STUDY DESIGN AND METHODS: IAT was performed using two different multispecific sera containing anti-Fr(a) and a series of multispecific sera containing antibodies against other blood group antigens of the Diego blood group system but not agglutinating Fr(a+) RBCs. Biosynthesis of band 3 was studied by RT-PCR of reticulocyte RNA and electrophoresis of solubilized RBC membranes. Anion exchange function of band 3 was studied by measuring the influx of radiolabeled sulfate. RESULTS: RBCs from the Glu480Lys carrier were agglutinated with sera containing anti-Fr(a) and not by sera with specificities for other antigens of the Diego system. We detected identical quantities of mRNA corresponding to the two band-3 alleles and normal content of band 3 in the RBC membranes, as well as normal sulfate influx into RBCs from the Fr(a) heterozygote. CONCLUSIONS: We confirmed the previously reported molecular basis of the Fr(a) antigen, thus providing supportive evidence for the existence of the second extracellular loop of band 3. We also demonstrated that this substitution does not affect mRNA stability, surface expression, and anion exchange function of band 3.

Amino Acid Sequence↗
PubMed2005

Profile of Toll-like receptor expressions and induction of nitric oxide synthesis by Toll-like receptor agonists in chicken monocytes.

Toll-like receptors (TLRs) play a major role in the innate immune system for initial recognition of microbial pathogens and pathogen associated components. Nitric oxide (NO) is generated in immune cells in response to microbial stimulation and is involved in pathogenesis and control of infection. We used RT-PCR analysis to examine the TLR expression profile on chicken monocytes and demonstrated these cells express chicken TLR2, 3, 4, 6, and 7. TLR5 was not detected by the TR-PCR. We also investigated the differential induction of NO synthesis in chicken monocytes by TLR agonists, including flagellin (FGN, from Salmonella typhimurium), synthetic lipoprotein Pam3CSK4 (PAM), lipopolysaccharide (LPS, from Salmonella enteritidis), lipoteichoic acid (LTA, from Staphylococcus aureus), the synthetic double stranded RNA analog (poly I:C), the guanosine analog, loxoribine (LOX), and synthetic CpG oligodeoxydinucleotide (CpG-ODN). Our results indicate that there was a vast difference among these agonists for their ability to induce NO production. CpG-ODN and LPS were the most potent stimuli and induced significant quantities of NO in cultured monocytes, whereas LTA stimulated significant NO production only at high concentrations. Other agonists such as FGN and poly I:C stimulated very little NO, while PAM, LOX, and nCpG-ODN (control ODN) did not induce NO production. RT-PCR analysis demonstrated that LPS, LTA, and CpG-ODN induced inducible nitric oxide synthase (iNOS) expression in monocytes; whereas the other agonists did not. The presence of TLRs on chicken monocytes and the differential induction of NO production in chicken monocytes by various TLR agonists suggest the differentiation of signaling pathways downstream of individual TLRs.

Animals↗
PubMed2004

Distance-restrained docking of rifampicin and rifamycin SV to RNA polymerase using systematic FRET measurements: developing benchmarks of model quality and reliability.

We are developing distance-restrained docking strategies for modeling macromolecular complexes that combine available high-resolution structures of the components and intercomponent distance restraints derived from systematic fluorescence resonance energy transfer (FRET) measurements. In this article, we consider the problem of docking small-molecule ligands within macromolecular complexes. Using simulated FRET data, we have generated a series of benchmarks that permit estimation of model accuracy based on the quantity and quality of FRET-derived distance restraints, including the number, random error, systematic error, distance distribution, and radial distribution of FRET-derived distance restraints. We find that expected model accuracy is 10 A or better for models based on: i), > or =20 restraints with up to 15% random error and no systematic error, or ii), > or =20 restraints with up to 15% random error, up to 10% systematic error, and a symmetric radial distribution of restraints. Model accuracies can be improved to 5 A or better by increasing the number of restraints to > or =40 and/or by optimizing the distance distribution of restraints. Using experimental FRET data, we have defined the positions of the binding sites within bacterial RNA polymerase of the small-molecule inhibitors rifampicin (Rif) and rifamycin SV (Rif SV). The inferred binding sites for Rif and Rif SV were located with accuracies of, respectively, 7 and 10 A relative to the crystallographically defined binding site for Rif. These accuracies agree with expectations from the benchmark simulations and suffice to indicate that the binding sites for Rif and Rif SV are located within the RNA polymerase active-center cleft, overlapping the binding site for the RNA-DNA hybrid.

Binding Sites↗
PubMed1996

Cloning and quantification of galanin-1 receptor expression by mucosal cells lining the human gastrointestinal tract.

Recent studies suggest that galanin receptors may be expressed by mucosal epithelial cells lining the GI tract and play a role in regulating ion transport. However, no information exists as to which receptor subtype is expressed by these mucosal cells, or the relative distribution of such receptors within the GI tract. In this study we cloned the human galanin-1 receptor (huGal 1-R) from small intestinal RNA, and demonstrate its expression in endoscopically obtained mucosal pinch biopsies by RT-PCR from the esophagus to the rectum. Semi-quantitative PCR (SQ-PCR) was performed and revealed the largest amount of huGal 1-R message in the duodenum and the least amount in the gastric fundus. This study for the first time directly demonstrates the presence and quantity of huGal 1-R message in human G1 tract mucosal epithelial cells, and suggests that this subtype is of primary importance in regulating galanin-induced changes in intestinal absorption.

Base Sequence↗
PubMed1996

Muscle and liver protein synthesis adapt efficiently to food deprivation and refeeding in 12-month-old rats.

Our aim was to analyze mechanisms involved in the adaptation of protein metabolism to food deprivation and refeeding in adult rats. Twelve-month-old rats, which had been food-deprived for 113 h and refed for 6 h, were injected subcutaneously with a flooding dose of valine (with 50% [1-13C]-L-valine) to measure in vivo protein synthesis in tibialis anterior, soleus and liver. Protein and RNA contents were also measured. In both muscles, protein mass was maintained during food deprivation. Due to a drop in protein synthetic capacity (Cs), total and myofibrillar protein synthesis rates were reduced in food-deprived rats and were not stimulated by a 6-h refeeding. In contrast, protein levels were maintained lower than RNA levels in liver during food deprivation, and Cs was higher than in fed rats. Protein synthesis rates and ribosomal efficiency were reduced in food-deprived rats. Due to maintenance of protein synthetic capacity, there was a rapid stimulation of liver protein synthesis with refeeding, which induced a significant rise in protein mass (also related to an inhibition of protein degradation). In conclusion, coordinated responses of liver and muscles allowed a sparing of muscle proteins during food deprivation and a rapid recovery of liver proteins during refeeding. Control of ribosome quantity could play a critical role in these adaptations in tissue protein synthesis in adult rats.

Aging↗
PubMed2000

Marked differences in quantity of infectious human immunodeficiency virus type 1 detected in persons with controlled plasma viremia by a simple enhanced culture method.

Culture of autologous CD4 lymphocytes from peripheral blood mononuclear cells compared favorably with two other methods for the measurement of cell-associated human immunodeficiency virus type 1 (HIV-1). For subjects with undetectable HIV-1 RNA levels in plasma, there was a 10,000-fold range of cell-associated virus detected. This method provides a simple and reproducible means for monitoring cell-associated HIV-1.

CD4-Positive T-Lymphocytes↗
PubMed1972

Procedure for the preparation of milligram quantities of adenovirus messenger ribonucleic acid.

Late after adenovirus 2 infection (18 hr), nearly all newly synthesized polysomal messenger ribonucleic acid (mRNA) is viral specified. Large amounts of adenovirus mRNA have been purified by utilizing membrane filtration at high ionic strength. With this procedure, molecules that contain polyadenylic acid [poly (A)] tracts are bound selectively, and ribosomal RNA can be separated from the viral mRNA which contains poly(A). Polysomal RNA synthesized 18 hr after infection was labeled in the presence of 0.02 mug of actinomycin D per ml and extracted at pH 9.0. This RNA annealed 40% to 3 mug of adenovirus 2 deoxyribonucleic acid; the RNA selected by membrane filtration bound 80% under the same conditions. The RNA eluted from membrane filters was 80 to 90% greater than 18S and contained species migrating as 31, 27, and 24S. Binding of polysomal RNA to individual membrane filters was linear, using as much as 300 mug of RNA per membrane. A 1.1-mg amount of viral RNA was prepared from 17.7 mg of polysomal RNA that had been purified by extraction at pH 9.0.

Adenoviridae↗
PubMed1983

Splicing of adenovirus 2 early region 1A mRNAs is non-sequential.

The r-strand of early region 1A (E1A) of adenovirus serotype 2 is transcribed into three completely overlapping messenger RNA species, a 9S, a 12S and a 13S mRNA. These three mRNAs are processed from a common colinear RNA precursor and differ only with regard to the size of the intron removed during mRNA maturation. We have studied the processing pathways for the E1A mRNAs by using an assay for transient expression of recombinant plasmids containing the E1A region. All three region E1A mRNAs are synthesized and transported to the cytoplasm in sufficient quantities to permit a detailed study of their structure by S1 endonuclease analysis and primer extension. Additionally, we show that the 72 base-pair repeat from simian virus 40 (SV40), when located upstream of the E1A promoter stimulates expression of the E1A mRNAs five- to tenfold. In order to determine whether splicing of the E1A mRNAs is sequential, i.e. whether the 13S and 12S mRNAs can serve as intermediates in splicing, we constructed two plasmids that lack the intervening sequences that are removed during the maturation of the 12S and 13S mRNAs, respectively. From an analysis of the RNAs produced after transfection with these deletion mutants, the following major conclusions can be made. (1) Splicing of the E1A mRNAs is non-sequential, i.e. the 13S, 12S and 9S RNAs are generated by separate splicing events using the nuclear colinear transcript as the only precursor RNA. (2) RNA splicing is not a prerequisite for an efficient transport of the E1A mRNAs to the cytoplasm. (3) The 13S RNA can be further processed to 12S and 9S RNA species. These splicing events are, however, illegitimate and give rise to 12S and 9S RNAs that both lack one nucleotide at the splice junction. (4) A coupling between splicing and nuclear transport is most likely required in vivo to prevent illegitimate splicing of the 13S mRNA. (5) The 12S RNA does not serve as a precursor for further processing to the 9S RNA. (6) Splicing of the E1A mRNAs followed strictly the G-T-A-G rule.

Adenoviruses, Human↗
PubMed1994

Effects of an antisense napin gene on seed storage compounds in transgenic Brassica napus seeds.

To manipulate the quantity and quality of storage components in Brassica napus seeds, we have constructed an antisense gene for the storage protein napin. The antisense gene was driven by the 5'-flanking region of the B. napus napin gene to express antisense RNA in a seed-specific manner. Seeds of transgenic plants with antisense genes often contained reduced amounts of napin. In some transgenic plants, no accumulation of napin was observed. However, the total protein content of transgenic and wild-type seeds did not differ significantly. Seeds lacking napin accumulated 1.4 to 1.5 times more cruciferin than untransformed seeds, although the oleosin content was not affected. Fatty acid content and composition in the seeds of transgenic plants were also analyzed by gas chromatography. Though the total fatty acid content of the transformants was the same as that of non-transformants, there was a reduction in 18:1 contents and a concomitant increase of 18:2 in seeds with reduced napin levels. This observed change in fatty acid composition was inherited in the next generation.

2S Albumins, Plant↗
PubMed2006

Sparing of methionine requirements: evaluation of human data takes sulfur amino acids beyond protein.

The intimate relation between amino acids and protein and nitrogen requirements is well recognized. Nutrition research has focused on the capacity of food to meet the need for nitrogen and indispensable amino acids (IAA) and led to the conclusion that the quality, not just the quantity, of protein is critical. This is especially relevant in regard to the sulfur amino acids (SAA) methionine and cysteine because of the increased understanding of their relation to chronic diseases (e.g., cardiovascular disease, dementia, cirrhosis), immunomodulation, DNA transcription, and RNA translation. Considerable effort has been expended to determine whether and to what extent cysteine can spare the requirement for the IAA methionine. In vivo studies in humans generally concur that the dietary requirement of the SAA ranges between 13 and 16 mg.kg(-1).d(-1), but how much can be met by cysteine relative to methionine remains controversial. This review examines the current status of in vivo estimates of methionine and cysteine requirements in human adults and considers needs beyond what is necessary for protein synthesis. Factors influencing the utilization of methionine and cysteine, especially those conditions that lead to toxicity on the one hand or beneficial effects on the other, are discussed. Data on alternative dietary sources of methyl groups (e.g., betaine, choline, phosphatidylcholine, S-adenosylmethionine, S-methylmethionine) or sulfur (e.g., N-acetylcysteine or L-2-oxothiazolidine-4-carboxylic acid) support a role for the SAA "beyond protein." Other pathways may influence the specific requirement for methionine and/or cysteine, especially when the person is challenged by disease, inadequate availability of food, or environmental stress.

Amino Acids, Essential↗
PubMed1998

Changes in expression of contractile FP and relaxatory EP2 receptors in pregnant rat myometrium during late gestation, at labor, and postpartum.

Prostaglandins synthesized at parturition may act via specific myometrial receptors as mediators of uterine contractions. Several isoforms of eicosanoid (prostaglandin) receptors, identified by pharmacological means, are linked to contractile (FP, EP1, EP3) or relaxatory (EP2, EP4, IP, DP) intracellular pathways. Changes in mRNA expression of the contractile FP and the relaxatory EP2 receptor were measured in myometrium throughout gestation, at parturition, and postpartum. Timed pregnant rats were killed at 0900 h on Day 16, 18, 20, 21, 21.5, or 22 (parturition) of pregnancy or one day postpartum (n = 5 animals/group). A longitudinal section of myometrium was removed, total RNA was extracted, and semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed for glyceraldehyde phosphate dehydrogenase (GAPDH), calponin, EP2, and FP receptor mRNA expression. Complementary DNA products were run on agarose gels and visualized, and the quantity of cDNA product was measured against a DNA mass ladder. RT-PCR product identity was confirmed by restriction enzyme cleavage. EP2 receptor mRNA expression was highest at Day 16 and declined significantly to Day 21.5 and one day postpartum (p < 0.05, Student-Newman-Keuls procedure). Expression of FP receptor mRNA was low at Day 16 of gestation and increased significantly until delivery (p < 0.05, ANOVA) at Day 22, then fell to prepartum levels at one day postpartum. Myometrial activity at parturition may change from an active quiescent to an active contractile state in concert with a decline in expression of the relaxatory EP2 receptors and up-regulation of contractile FP receptors.

Animals↗
PubMed1986

Molecular cloning of cDNA for rat argininosuccinate lyase and its expression in rat hepatoma cell lines.

Using antibody and plaque hybridization screening, we isolated rat argininosuccinate lyase (AS lyase) cDNA clones from a liver cDNA library prepared in the phage expression vector lambda gt11. Five overlapping cDNAs covering 1.7 kilobases of the estimated 2.0-kilobase AS lyase mRNA were characterized and confirmed as AS lyase sequences by hybrid selection. We examined the differential expression of AS lyase in rat liver and four rat hepatoma cell lines (7800C1, H4, HTC, and MH1C1). These cells exhibited a 60-fold range of AS lyase enzyme activity, with a direct correlation between activity, amount of AS lyase immunoreactive protein, and quantity of specific AS lyase mRNA. These observations suggest that the differences in AS lyase expression between rat liver and the hepatoma cell lines result from variations in AS lyase transcriptional activity or alterations in nuclear processing of AS lyase RNA.

Animals↗
PubMed1983

Effects of dietary fiber on mucosal growth and cell proliferation in the small intestine of the rat: a comparison of oat bran, pectin, and guar with total fiber deprivation.

The effects of dietary fiber on small intestinal mucosal mass, morphology, and cytokinetics were studied by feeding three qualitatively different forms of fiber to 40 rats for 4 wk. A fiber-free diet was fed to the control group and a similar diet with either a 20% oat bran, 10% pectin, or 10% guar supplement to the other three groups. All groups of rats exhibited similar caloric intakes and weight gains. Only the guar diet produced a significant increase in mucosal mass, as demonstrated by 19.1% increase in mucosal weight (p less than 0.05), a 16.7% increase in RNA (p less than 0.05) and an increase in DNA from 38.3 +/- 1.8 (SEM) to 44.6 +/- 2.3 micrograms/cm intestine/100 g body weight (p less than 0.05) when compared to the controls. The pectin-diet produced a decrease in villus height from 102.1 +/- 2.4 to 94.3 +/- 2.4 cells but an increase in crypt column length from 25.3 +/- 0.6 to 27.4 +/- 0.4 cells when compared to the controls (p less than 0.05). The shift in labeling index distribution curves for pectin and guar to the right and for oat bran to the left indicated an increase and decrease respectively in labeling index. The higher rate of epithelial cell migration in the pectin- and guar-fed groups shortened their estimated villus cell transit times to 36.4 +/- 0.7 and 37.0 +/- 1.4 h, respectively, when compared with 42.6 +/- 1.2 h in the oat bran and 41.1 +/- 1.0 h in the control (p less than 0.05). These results show that the modulation of small intestinal mucosal structure and growth by dietary fiber appears to be mediated through alterations in cell proliferation and that these changes depend not only on the quantity but also the quality of the fiber present in the diet.

Animals↗