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Biomedical subjects

J B Russell

Publications and source records attributed to J B Russell.

At least 73 records · Page 4Linked to original sources

The effect of amino nitrogen on the energetics of ruminal bacteria and its impact on energy spilling.

The predominant ruminal bacteria that were obtained from a 10(8) dilution of ruminal fluid could be maintained as a mixed population for long periods as long as the bacteria were provided with a complex mixture of carbohydrates. Growth of predominant ruminal bacteria in carbohydrate-limited, ammonia-excess, continuous cultures (0.07/h) had a low requirement for maintenance energy, but the nongrowth energy dissipation of ammonia-limited, carbohydrate-excess, predominant ruminal bacteria was approximately 10-fold higher (0.96 vs. 0.09 mg of hexose equivalent/mg of protein per h, respectively). Mathematical derivations indicated that this additional nongrowth energy dissipation could be accommodated by an energy spilling function that was independent of the growth rate. Peptides and amino acids had little impact on the yield of carbohydrate-limited, ammonia-excess, continuous cultures (0.07/ h), but amino N greatly increased the growth rate and yield of excess-energy batch cultures. The change in growth rate and yield that was dependent on amino N indicated that the energy-excess batch cultures had the same capacity to spill energy as did the ammonia-limited, carbohydrate-excess, predominant ruminal bacteria (0.80 vs. 0.86 mg of hexose equivalent/mg of protein per h, respectively). When the energy-excess batch cultures were provided with amino N, the growth rate increased, the difference in anabolic and catabolic rates was smaller, and less energy was spilled.

Amino Acids↗

How many ruminal bacteria are there?

With the development of strictly anaerobic techniques and habitat-simulating media, a variety of bacteria were isolated from the rumen in the 1940s and 1950s. Based on standard morphological and physiological characteristics, the microbial ecosystem of the rumen contains a very complex population of bacteria. In recent years, ruminal bacteria have been re-evaluated with newer, more objective, and genetically valid methods of classification. Ribosomes are complicated structures, and their DNA-encoding sequences are relatively free from selective pressure. Because ribosomes have evolved slowly, they provide a long-term natural history of evolution. The invariable and hypervariable regions of rRNA genes can be used to group bacteria into kingdoms, genera, and species. The 16S rRNA sequences have provided a basis for renaming some ruminal species (Bacteroides amylophilus is now Ruminobacter amylophilus and Bacteroides succinogenes is now Fibrobacter succinogenes) and for classifying at least one recently isolated ruminal bacterium (e.g., Clostridium aminophilum). The DNA:DNA hybridization is a more sensitive method of assessing bacterial relatedness than is 16S rRNA. Bacterial strains within a species should have a high degree of DNA:DNA homology, but some species of ruminal bacteria (e.g., Prevotella ruminicola and Butyrivibrio fibrisolvens) had highly unrelated strains. Studies of 16S rRNA and DNA:DNA hybridization indicate that the diversity of ruminal bacteria has been greatly underestimated. Traditional studies of phylogeny of ruminal bacteria were stymied by the fastidious growth requirements of many ruminal bacteria, and enumeration was tedious and inaccurate. Modern methods of bacterial classification do not require in vitro culture and have the potential of detecting even a single cell.

Animal Nutritional Physiological Phenomena↗

Why do many ruminal bacteria die and lyse so quickly?

Studies using 15N have indicated that as much as 50% of the microbial mass turns over before N passes to the lower gut, and this N recycling significantly decreases the availability of microbial protein. Protozoa digest bacteria and smaller protozoa, but bacterial protein can turn over even if protozoa are not present. Fibrobacter succinogenes cultures lyse even when they are growing, and the lysis rate is independent of growth rate. When extracellular sugar is depleted, F. succinogenes secretes an extracellular proteinase that inactivates the autolysins. This method of autolytic regulation decreases the turnover of stationary cells. Bacteriophage and anaeroplasma can cause lysogeny, but, as yet, there is little proof that these processes are important determinants of bacterial turnover in vivo. Dietary manipulations (e.g., salt feeding and particle size reduction) that increase liquid and solid dilution rates can increase bacterial flow by decreasing bacterial residence time and turnover. Some dead ruminal bacteria are able to maintain their cellular integrity, and the ratio of dead to live cells in ruminal fluid may be as great as 10:1. Bacterial survival appears to be at least partially explained by the method of sugar transport. When bacteria rely solely on mechanisms of ion-coupled sugar symport, an energized membrane is necessary for the reinitiation of growth. If group translocation (phosphotransferase system) is the mechanisms of transport, uptake can be driven by phosphoenolpyruvate, and an energized membrane and the storage of intracellular reserve materials are not an absolute criteria for survival. In some cases, N deprivation accelerates death. When Prevotella ruminicola was limited for N under conditions of excess energy, methylglyoxal production caused a rapid decrease in viability. The impact of bacterial death in the rumen is not clear-cut. If the rate of fermentation is zero-order with respect to cell concentration (substrate-limited), cell death would have little impact on digestion.

Animals↗

Why are ruminal cellulolytic bacteria unable to digest cellulose at low pH?

Ruminant animals depend on cellulolytic ruminal bacteria to digest cellulose, but these bacteria cannot resist the low ruminal pH that modern feeding practices can create. Because the cellulolytic bacteria cannot grow on cellobiose at low pH, pH sensitivity is a general aspect of growth and not just a limitation of the cellulases per se. Acid-resistant ruminal bacteria have evolved the capacity to let their intracellular pH decrease, maintain a small pH gradient across the cell membrane, and prevent an intracellular accumulation of VFA anions. Cellulolytic bacteria cannot grow with a low intracellular pH, and an increase in pH gradient leads to anion toxicity. Prevotella ruminicola cannot digest native cellulose, but it grows at low pH and degrades the cellulose derivative, carboxymethylcellulose. The Prevotella ruminicola carboxymethylcellulase cannot bind to cellulose, but a recombinant enzyme having the Prevotella ruminicola catalytic domain and a binding domain from Thermomonspora fusca was able to bind and had cellulase activity that was at least 10-fold higher. Based on these results, gene reconstruction offers a means of converting Prevotella ruminicola into a ruminal bacterium that can digest cellulose at low pH.

Animals↗

A review of the genetics of recurrent pregnancy loss.

The field of recurrent pregnancy loss (RPL) has seen a blossoming of investigations in the etiology and treatment of this condition over the last few years. Despite all of this activity, there remains much controversy in all aspects of the field. This ranges from discrepancies in the definition of recurrent pregnancy loss to elements in the work-up and the very existence of certain conditions for which aggressive therapeutic modalities are proposed. We will discuss the genetic component of RPL which is one of the few etiologies for RPL which has achieved near universal acceptance.

Abortion, Spontaneous↗

The effect of carbohydrates on the expression of the Prevotella ruminicola 1,4-beta-D-endoglucanase.

The beta-1,4-endoglucanase of the ruminal bacterium, Prevotella ruminicola B14, hydrolysed carboxymethylcellulose and barley glucan but not xylan or mannan. Endoglucanase activity was present in 88- and 82-kDa proteins, and there was at least a 20-fold variation in endoglucanase activity when P. ruminicola B14 was grown on different sugars. The highest activities were observed with mannose, cellobiose or xylose and little activity was observed with sucrose, arabinose or rhamnose, P. ruminicola B14 also had significant xylanase and mannanase activities, but these activities were present in proteins that had lower molecular masses than the endoglucanase and these proteins did not cross-react with antibody made against the endoglucanase. Mannanase activity has a similar pattern of expression to the endoglucanase, while the xylanase was not induced or repressed by the same sugars or combinations of sugars. The xylanase activity was greatest when xylan was the energy source for growth, but xylose was a very poor inducer of xylanase activity.

Animals↗

High success with gestational carriers and oocyte donors using synchronized cycles.

PURPOSE: Our purpose was to determine if exogenous hormone stimulation of the endometrium improves the implantation and pregnancy rate with a host uterus or gestational carrier. RESULTS: Nine couples underwent 14 embryo transfer cycles with a gestational carrier. The median age was 31.3 years for the infertile women donating the oocytes and 36.6 years for the gestational carriers. Eight pregnancies were achieved documented by ultrasound confirmation of cardiac activity. The pregnancy rate was 64.3%, with 1.6 cycles per patient. CONCLUSION: We show that synchronized exogenous hormone stimulation of the gestational carrier improves the success rate of embryo transfer. When the success rate is compared to a timed embryo transfer with the gestational carrier's natural cycle, from previous reported studies, exogenous hormonal stimulation appears to enhance endometrial receptivity and improve the implantation rate.

Adult↗

Retrospective evaluation of admission criteria for paediatric electrical injuries.

In the medical community, the practice of admitting all electrical burns for 24-48 h of observation, monitoring and laboratory evaluation is widespread. This retrospective review of paediatric electrical burns was conducted to determine which patients may safely be treated as outpatients. Retrospective analysis of all paediatric burns admitted between 1980 and 1991 identified 35 patients with electrical injuries. Patients were divided into two groups for analysis: those burned by exposure to household voltages (120-240 V; n = 26) and those exposed to high voltages, in excess of 1000 V (n = 9). The majority of household electrical injuries occurred secondary to contact with the household 120 V (21/26). Contact with an extremity accounted for the largest number of these injuries (18/26). The mouth was the second most frequent site of injury (7/26). Most of these patients (20/26) had < 1 per cent BSA burn. No patient in the household-voltage group had an arrythmia that required treatment, nor were there any identified examples of compartment syndrome or other vascular complications. Seven patients did require minimal skin grafting. No deaths occurred in either group. The patients in the household-voltage group were significantly younger. High-voltage electrical injuries occurred in an older patient population and required more aggressive care and surgical intervention. This was evident at the time of initial evaluation. Based on these data, healthy children with small partial-thickness electrical burns and no initial evidence of cardiac or neurovascular injury do not appear to need hospital admission.

Adolescent↗

Regulation of the lactose phosphotransferase system of Streptococcus bovis by glucose: independence of inducer exclusion and expulsion mechanisms.

Streptococcus bovis had a diauxic pattern of glucose and lactose utilization, and both of these sugars were transported by the sugar phosphotransferase system (PTS). Lactose catabolism was inducible, and S. bovis used the tagatose pathway to ferment lactose. Since a mutant that was deficient in glucose PTS activity transported lactose as fast as the wild-type, it appeared that S. bovis has separate enzyme IIs for glucose and lactose. The nonmetabolizable glucose analogue 2-deoxyglucose (2-DG) was a noncompetitive inhibitor of methyl beta-D-thiogalactopyranoside (TMG) transport, and cells that were provided with either glucose or 2-DG were unable to transport TMG or lactose. Because the glucose-PTS-deficient mutant could ferment glucose, but could not exclude TMG, it appeared that enzyme IIGlc rather than glucose catabolism per se was the critical feature of inducer exclusion. Cells that had accumulated TMG as TMG 6-phosphate expelled free TMG when glucose was added, but 2-DG was unable to cause TMG expulsion. The glucose-PTS-deficient mutant could still expel TMG in the presence of exogenous glucose. Membrane vesicles also exhibited glucose-dependent TMG exclusion and TMG expulsion. Membrane vesicles that were electroporated with phosphoenolpyruvate (PEP) and HPr retained TMG for more than 3 min, but vesicles that were electroporated with PEP plus HPr and fructose 1,6-diphosphate (FDP) (or glycerate 2-phosphate) lost their ability to retain TMG. Because FDP was able to trigger the ATP-dependent phosphorylation of HPr, it appeared that inducer expulsion was mediated by an FDP-activated protein kinase.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Proteins↗

Cellodextrin efflux by the cellulolytic ruminal bacterium Fibrobacter succinogenes and its potential role in the growth of nonadherent bacteria.

When glucose or cellobiose was provided as an energy source for Fibrobacter succinogenes, there was a transient accumulation (as much as 0.4 mM hexose equivalent) of cellobiose or cellotriose, respectively, in the growth medium. Nongrowing cell suspensions converted cellobiose to cellotriose and longer-chain cellodextrins, and in this case the total cellodextrin concentration was as much as 20 mM (hexose equivalent). Because cell extracts of glucose- or cellobiose-grown cells cleaved cellobioise and cellotriose by phosphate-dependent reactions and glucose 1-phosphate was an end product, it appeared that cellodextrins were being produced by a reversible phosphorylase reaction. This conclusion was supported by the observation that the ratio of cellodextrins to cellodextrins with one greater hexose [n/(n + 1)] was approximately 4, a value similar to the equilibrium constant (Keq) of cellobiose phosphorylase (J. K. Alexander, J. Bacteriol. 81:903-910, 1961). When F. succinogenes was grown in a cellobiose-limited chemostat, cellobiose and cellotriose could both be detected, and the ratio of cellotriose to cellobiose was approximately 1 to 4. On the basis of these results, cellodextrin production is an equilibrium (mass action) function and not just an artifact of energy-rich cultural conditions. Cellodextrins could not be detected in low-dilution-rate, cellulose-limited continuous cultures, but these cultures had a large number of nonadherent cells. Because the nonadherent cells had a large reserve of polysaccharide and were observed at all stages of cell division, it appeared that they were utilizing cellodextrins as an energy source for growth.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The cellular location of Prevotella ruminicola beta-1,4-D-endoglucanase and its occurrence in other strains of ruminal bacteria.

Prevotella ruminicola B(1)4, TC1-1, TF1-3, and TS1-5 all produced immunologically cross-reacting 88- and 82-kDa carboxymethyl cellulases (CMCases). P. ruminicola 23, 118B, 20-63, and 20-78 had much lower CMCase activities, and Western blots (immunoblots) showed no cross-reaction with the B(1)4 CMCase antiserum. Fibrobacter succinogenes S85 and Selenomonas ruminantium HD4 and D produced CMCase, but these enzymes were smaller and did not cross-react with the B(1)4 CMCase antiserum. The B(1)4 CMCase antiserum inhibited the B(1)4, TC1-1, TF1-3, and TS1-5 CMCase activities and agglutinated these cells, but it had no effect on the other strains or species. On the basis of these results, the B(1)4 CMCase is a strain-specific enzyme that is located on the outside surface of the cells. P. ruminicola B(1)4 cultures, grown on sucrose, did not have significant CMCase activity, but these cells could bind purified 88- and 82-kDa CMCase but not 40.5-kDa CMCase. Because the 40.5-kDa CMCase is a fully active, truncated form of the CMCase, it appears that the N-terminal domain of the 88-kDa B(1)4 CMCase anchors the CMCase to the cells. Cells grown on cellobiose produced at least 10-fold more CMCase than the sucrose-grown cells, and the cellobiose-grown cells could only bind 15% as much CMCase as sucrose-grown cells. Virtually all of the CMCase activity of exponentially growing cultures was cell associated, but CMCase activity was eventually detected in the culture supernatant. On the basis of the observation that the 88-kDa CMCase was gradually converted to the 82-kDa CMCase when cultures reached the stationary phase without a change in specific activity, it appears that the 82-kDa protein is probably a proteolytic degradation product of the 88-kDa CMCase.

Animals↗

Properties of two sugar phosphate phosphatases from Streptococcus bovis and their potential involvement in inducer expulsion.

Streptococcus bovis possesses two sugar phosphate phosphatases (Pases). Pase I is a soluble enzyme that is inhibited by the membrane fractions from lactose-grown cells and is insensitive to activation by S46D HPr, an analog of HPr(ser-P) of the sugar phosphotransferase system. Pase II is a membrane-associated enzyme that can be activated 10-fold by S46D HPr, and it appears to play a role in inducer expulsion.

Bacterial Proteins↗

Energetics of bacterial growth: balance of anabolic and catabolic reactions.

Biomass formation represents one of the most basic aspects of bacterial metabolism. While there is an abundance of information concerning individual reactions that result in cell duplication, there has been surprisingly little information on the bioenergetics of growth. For many years, it was assumed that biomass production (anabolism) was proportional to the amount of ATP which could be derived from energy-yielding pathways (catabolism), but later work showed that the ATP yield (YATP) was not necessarily a constant. Continuous-culture experiments indicated that bacteria utilized ATP for metabolic reactions that were not directly related to growth (maintenance functions). Mathematical derivations showed that maintenance energy appeared to be a growth rate-independent function of the cell mass and time. Later work, however, showed that maintenance energy alone could not account for all the variations in yield. Because only some of the discrepancy could be explained by the secretion of metabolites (overflow metabolism) or the diversion of catabolism to metabolic pathways which produced less ATP, it appeared that energy-excess cultures had mechanisms of spilling energy. Bacteria have the potential to spill excess ATP in futile enzyme cycles, but there has been little proof that such cycles are significant. Recent work indicated that bacteria can also use futile cycles of potassium, ammonia, and protons through the cell membrane to dissipate ATP either directly or indirectly. The utility of energy spilling in bacteria has been a curiosity. The deprivation of energy from potential competitors is at best a teleological explanation that cannot be easily supported by standard theories of natural selection. The priming of intracellular intermediates for future growth or protection of cells from potentially toxic end products (e.g., methylglyoxal) seems a more plausible explanation.

Adenosine Triphosphate↗

Laparoscopic oophorectomy.

Laparoscopic oophorectomy is one of the most controversial and challenging areas of endoscopic surgery. Judicious preoperative evaluation along with careful patient selection can reduce the risk of operating on an unsuspected ovarian malignancy. Appropriate patient consent and planning should be obtained before the procedure, in case an ovarian neoplasia should be encountered. Although endoscopic pelvic biopsy, treatment and staging has been reported recently, the standard of care still appears to be by a laparoscopy. The removal of benign cystic teratomas of all sizes can be handled laparoscopically with the assistance of a retrieval bag or pouch. Reports of endoscopic treatment of ovarian remnant syndrome, androgen insensitivity syndrome, and even prophylactic oophorectomy are appearing in the literature. They have major benefits to patients including reduced cost, decreased hospitalization and time away from work, and with similar or lower complication rates compared with other modes of treatment. Pelviscopy or operative laparoscopy is gaining in popularity and acceptance as our experience and training improves.

Adnexal Diseases↗

Alternative strategies of 2-deoxyglucose resistance and low affinity glucose transport in the ruminal bacteria, Streptococcus bovis and Selenomonas ruminantium.

Streptococcus bovis and Selenomonas ruminantium grew in the presence of the glucose analog, 2-deoxyglucose (2-DG), but the cells no longer had high affinity glucose transport. In S. bovis, 2-DG resistance was correlated with a decrease in phosphoenolpyruvate (PEP)-dependent glucose phosphotransferase (PTS) activity. The 2-DG-selected S. bovis cells relied solely upon a low affinity, facilitated diffusion mechanism of glucose transport and a 2-DG-resistant glucokinase (ATP-dependent). The glucokinase activity of S. ruminantium was competitively inhibited by 2-DG, and the 2-DG selected cells continued to use PEP-dependent PTS as a mechanism of glucose transport. In this latter case, the 2-DG selected cells switched from a mannosephosphotransferase (enzyme II) that phosphorylated glucose, mannose, and 2-DG, but not alpha-methylglucose to a glucosephosphotransferase (enzyme II) that phosphorylated glucose and alpha-methylglucoside but not 2-DG or mannose. The glucosephosphotransferase (enzyme II) had a very low affinity for glucose and the transport kinetics were similar to the facilitated diffusion system of S. bovis.

Animals↗

Emendation of the description of Acidaminococcus fermentans, a trans-aconitate- and citrate-oxidizing bacterium.

Ruminal fluid which was enriched with trans-aconitate yielded a gram-negative diplococcus (strain AO) which was identified by 16S ribosomal DNA sequence analysis as Acidaminococcus fermentans. In contrast to the original description, the A. fermentans type strain and strain AO were found to utilize citrate as an energy source and to produce hydrogen and hydrogen sulfide. The descriptions of the genus and species are emended accordingly.

Aconitic Acid↗

Energy-spilling reactions of Streptococcus bovis and resistance of its membrane to proton conductance.

Glucose-excess cultures of Streptococcus bovis consumed glucose faster than the amount that could be explained by growth or maintenance, and nongrowing chloramphenicol-treated cells had a rate of glucose consumption that was 10-fold greater than the maintenance rate. Because N,N-dicyclohexylcarbodiimide, an inhibitor of the membrane-bound F1F0 ATPase, eliminated the nongrowth energy dissipation (energy spilling) without a decrease in ATP and the rate of energy spilling could be increased by the protonophore 3,3',4',5-tetrachlorosalicylanilide, it appeared that a futile cycle of protons through the cell membrane was responsible for most of the energy spilling. When the rate of energy spilling was decreased gradually with iodoacetate, there was only a small decrease in the phosphorylation potential (delta G'p) and the theoretical estimate of H+ per ATP decreased from 4.2 to 3.6. On the bases of this ratio of H+ to ATP and the rate of ATP production, the flux of protons (amperage) across the cell membrane was directly proportional to the rate of energy spilling. Amperage values estimated from delta G'p were, however, nearly twice as great as values which were estimated from the heat production (delta H) of the cells [amperage = (0.38 x wattage)/delta p]. The last comparison indicated that only a fraction of the delta G of ATP hydrolysis was harvested by the F1F0 ATPase to pump protons. Both estimates of amperage indicated that the resistance of the cell membrane to proton conductance was inversely proportional to the log of the energy-spilling rate.

Adenosine Triphosphate↗