Association between childhood cognitive skills & adult suicidal behavior: A systematic review and meta-analysis.

BACKGROUND: It is unclear whether cognitive skill deficits during childhood carry risk for suicide attempt or mortality later in adulthood at the population level. We conducted a systematic review and meta-analysis of population-based studies examining the association between childhood cognitive skills and adult suicidal behavior, namely attempt and mortality. METHOD: We systematically searched databases for articles then extracted study characteristics and estimates on the association between childhood cognitive skills (i.e., IQ or school performance at age ≤ 18 years) and later suicide attempt and mortality. Random-effect meta-analysis was used to quantify this association across all studies with available data. RESULTS: Twenty-three studies met the inclusion criteria and suggest an association between lower childhood cognitive skills and increased risk of suicidal behavior. Meta-analysis of the adjusted estimates from 11 studies (N = 2,830,191) found the association to be small but statistically significant. Heterogeneity was significant but moderate, and results were unlikely to be influenced by publication bias. In subgroup analyses, associations were significant only for males. No difference in effect size was found between suicide attempt and suicide mortality. LIMITATIONS: Cognitive skills were measured with different cognitive subtests. Heterogeneity in the age of cognitive skills assessment. Meta-regression and subgroup analyses were based on a relatively low number of studies. CONCLUSIONS: Individuals with lower cognitive skills in childhood have a greater risk of suicidal behavior in adulthood, especially males. Although the association was small, interventions improving cognitive skills may yield large effects on suicide prevention at the population level if the association is causal.

Low MoCA performances correlate with suicidal ideation in late-life depression.

Late-life depression remains an underdiagnosed clinical entity, mainly because the presence of cognitive impairment in the elderly leads clinicians to suspect dementia rather than depression. Our objective was to analyze the cognitive abilities of elderly depressed patients using the Montreal Cognitive Assessment (MoCA) in relation to the presence or absence of suicidal ideation. The MoCA, Beck Scale of Suicidal Ideation, Hamilton Anxiety Scale, and Hamilton Depression Scale were administered to 72 patients with a recent history of late life depression: 43 with suicidal ideation and 29 non-suicidal controls. The results show that suicidal patients demonstrated significantly worse performance on the MoCA total score and the delayed recall subtest in comparison to non-suicidal controls. In addition, after adjusting for age and depression, poorer performance on the MoCA total score correlated to the presence of suicidal ideation. We found that the MoCA total score is able to predict the presence of suicidal ideation in depressed elderly patients in a fair-to-good manner. As late-life depression is already established as a potential prodrome of dementia, longitudinal follow-up may determine whether depressed individuals with suicidal ideation are at higher risk of converting to dementia.

Association between hypovitaminosis D and cognitive inhibition impairment during major depression episode.

BACKGROUND: Major depressive episode (MDE) has been associated with cognitive functioning alteration and hypovitaminosis D (hypoVD), but the relationship between hypoVD, depression, and cognition is not well understood. We aimed to compare patient with MDE with or without hypoVD in regard of cognitive functioning. METHODS: 91 patients (38.5 years old, 65.9% female) with MDE were included in a cross-sectional study and were evaluated with a complete cognitive battery. None of the participants were medicated at the time of the inclusion. Serum 25-hydroxyvitamin D was measured using LC-MS/MS method, and hypovitaminosis was defined as 25OHD < 50nmol/L. Covariates were gender, season of dosage, first MDE onset, age, body mass index and depression severity RESULTS: Patients with hypoVD demonstrated a higher stroop intereference index time underscoring that means low cognitive inhibition ability. Mutiple logistic regression confirmed that hypoVD was significantly associated with high stroop interference time index after controlling by gender, season of dosage, first MDE onset, age, body mass index and depression severity. CONCLUSION: Our results suggest that patient with MDE having hypoVD may be more prone to cognitive impairment.

Narrow hybrid zones in spite of very low population differentiation in neutral markers in an island bird species complex.

Patterns of phenotypic and genic frequencies across hybrid zones provide insight into the origin and evolution of reproductive isolation. The Reunion grey white-eye, Zosterops borbonicus, exhibits parapatrically distributed plumage colour forms across the lowlands of the small volcanic island of Reunion (Mascarene archipelago). These forms meet and hybridize in regions that are natural barriers to dispersal (rivers, lava fields). Here, we investigated the relationship among patterns of differentiation at neutral genetic (microsatellite) markers, phenotypic traits (morphology and plumage colour) and niche characteristics across three independent hybrid zones. Patterns of phenotypic divergence revealed that these hybrid zones are among the narrowest ever documented in birds. However, the levels of phenotypic divergence stand in stark contrast to the lack of clear population neutral genetic structure between forms. The position of the hybrid zones coincides with different natural physical barriers, yet is not associated with steep changes in vegetation and related climatic variables, and major habitat transitions are shifted from these locations by at least 18 km. This suggests that the hybrid zones are stabilized over natural dispersal barriers, independently of environmental boundaries, and are not associated with niche divergence. A striking feature of these hybrid zones is the very low levels of genetic differentiation in neutral markers between forms, suggesting that phenotypic divergence has a narrow genetic basis and may reflect recent divergence at a few linked genes under strong selection, with a possible role for assortative mating in keeping these forms apart.

The role of selection and historical factors in driving population differentiation along an elevational gradient in an island bird.

Adaptation to local environmental conditions and the range dynamics of populations can influence evolutionary divergence along environmental gradients. Thus, it is important to investigate patterns of both phenotypic and genetic variations among populations to reveal the respective roles of these two types of factors in driving population differentiation. Here, we test for evidence of phenotypic and genetic structure across populations of a passerine bird (Zosterops borbonicus) distributed along a steep elevational gradient on the island of Réunion. Using 11 microsatellite loci screened in 401 individuals from 18 localities distributed along the gradient, we found that genetic differentiation occurred at two spatial levels: (i) between two main population groups corresponding to highland and lowland areas, respectively, and (ii) within each of these two groups. In contrast, several morphological traits varied gradually along the gradient. Comparison of neutral genetic differentiation (FST ) and phenotypic differentiation (PST ) showed that PST largely exceeds FST at several morphological traits, which is consistent with a role for local adaptation in driving morphological divergence along the gradient. Overall, our results revealed an area of secondary contact midway up the gradient between two major, cryptic, population groups likely diverged in allopatry. Remarkably, local adaptation has shaped phenotypic differentiation irrespective of population history, resulting in different patterns of variation along the elevational gradient. Our findings underscore the importance of understanding both historical and selective factors when trying to explain variation along environmental gradients.

Psychological characteristics of patients with myotonic dystrophy type 1.

OBJECTIVES: Myotonic dystrophy type 1 (DM1) is the most common adult-onset muscular dystrophy. It is associated with motor symptoms but patients also display non-motor symptoms such as particular personality traits. Studies have reported mixed results about personality characteristics which may be attributable to small sample sizes, different disease severity of groups studied, and use of different questionnaires or method. This study aimed to describe the psychological characteristics of a large cohort of patients with DM1, to characterize those at risk of developing a psychiatric disorder, and to compare characteristics between two DM1 phenotypes, a mild and more severe adult-onset phenotype. METHODS: Two hundred patients with DM1 (152 adult-onset; 48 mild) were asked to complete questionnaires assessing personality traits, psychological symptoms, self-esteem, and suicidal risk. Neurological and neuropsychological assessments were performed to compare personality characteristics to clinical and cognitive measures. RESULTS: Patients with DM1 globally showed personality traits and psychological symptoms in the average range compared to normative data, with normal levels of self-esteem and suicidal ideation. However, 27% of patients were found to be at high risk of developing a psychiatric disorder. Moreover, psychological traits differed across phenotypes, with the most severe phenotype tending to show more severe psychological symptoms. The presence of higher phobic anxiety and lower self-esteem was associated with lower education, a higher number of CTG repeats, more severe muscular impairment, and lower cognitive functioning (P < 0.001). CONCLUSIONS: Different phenotypes should thus be taken into account in clinical settings for individual management of patients and optimizing therapeutic success.

Extremely reduced dispersal and gene flow in an island bird.

The Réunion grey white-eye, Zosterops borbonicus, a passerine bird endemic to Réunion Island in the Mascarene archipelago, represents an extreme case of microgeographical plumage colour variation in birds, with four distinct colour forms occupying different parts of this small island (2512 km(2)). To understand whether such population differentiation may reflect low levels of dispersal and gene flow at a very small spatial scale, we examined population structure and gene flow by analysing variation at 11 microsatellite loci among four geographically close localities (<26 km apart) sampled within the distribution range of one of the colour forms, the brown-headed brown form. Our results revealed levels of genetic differentiation that are exceptionally high for birds at such a small spatial scale. This strong population structure appears to reflect low levels of historical and contemporary gene flow among populations, unless very close geographically (<10 km). Thus, we suggest that the Réunion grey white-eye shows an extremely reduced propensity to disperse, which is likely to be related to behavioural processes.

H2O vapor transmission rate through polyethylene naphthalate polymer using the electrical Ca test.

The electrical Ca test was used to measure H2O vapor transmission through polyethylene naphthalate (PEN) polymer with a thickness of 200 µm. On the basis of the time required for the normalized conductance of the Ca film to reach zero, the H2O vapor transmission rate was determined versus H2O flux, temperature, and saturation of the PEN polymer with H2O. The H2O vapor transmission rate was proportional to the H2O flux and only weakly dependent on temperature at constant H2O flux. The transmission coefficient, Γ, for H2O through the PEN polymer at 70 °C was Γ âˆ¼ 3.2 × 10(-10). The corresponding water vapor transmission rate (WVTR) at 70 °C/80% RH was 0.65 g/(m(2) day). The temperature dependence of the H2O vapor transmission rate through PEN at constant H2O flux yielded an activation barrier of E = 12.4 kJ/mol. There was no observable reservoir effect for H2O in the PEN polymer. The H2O vapor transmission rates for initially dry or H2O-saturated PEN polymer substrates were nearly identical at various temperatures. Although the time required for the normalized conductance of the Ca film to reach zero was inversely proportional to the H2O flux, the Ca film conductance did not decrease linearly versus H2O exposure. The Ca film conductance changed very little during initial H2O exposure. This behavior may be caused by the nonlinear oxidation kinetics of the Ca film.

Offspring from mothers fed a 'junk food' diet in pregnancy and lactation exhibit exacerbated adiposity that is more pronounced in females.

We have shown previously that a maternal junk food diet during pregnancy and lactation plays a role in predisposing offspring to obesity. Here we show that rat offspring born to mothers fed the same junk food diet rich in fat, sugar and salt develop exacerbated adiposity accompanied by raised circulating glucose, insulin, triglyceride and/or cholesterol by the end of adolescence (10 weeks postpartum) compared with offspring also given free access to junk food from weaning but whose mothers were exclusively fed a balanced chow diet in pregnancy and lactation. Results also showed that offspring from mothers fed the junk food diet in pregnancy and lactation, and which were then switched to a balanced chow diet from weaning, exhibited increased perirenal fat pad mass relative to body weight and adipocyte hypertrophy compared with offspring which were never exposed to the junk food diet. This study shows that the increased adiposity was more enhanced in female than male offspring and gene expression analyses showed raised insulin-like growth factor-1 (IGF-1), insulin receptor substrate (IRS)-1, vascular endothelial growth factor (VEGF)-A, peroxisome proliferator-activated receptor-gamma (PPARgamma), leptin, adiponectin, adipsin, lipoprotein lipase (LPL), Glut 1, Glut 3, but not Glut 4 mRNA expression in females fed the junk food diet throughout the study compared with females never given access to junk food. Changes in gene expression were not as marked in male offspring with only IRS-1, VEGF-A, Glut 4 and LPL being up-regulated in those fed the junk food diet throughout the study compared with males never given access to junk food. This study therefore shows that a maternal junk food diet promotes adiposity in offspring and the earlier onset of hyperglycemia, hyperinsulinemia and/or hyperlipidemia. Male and female offspring also display a different metabolic, cellular and molecular response to junk-food-diet-induced adiposity.

The effect of carbohydrate source on nitrogen capture in dairy cows on pasture.

The objective of this study was to determine if feeding carbohydrate supplements with faster degradation rates than corn to dairy cows grazing ryegrass would improve nitrogen capture, milk production, and components. Treatments were grain supplements based on: 1) corn (CORN), 2) barley and molasses (BM), or 3) citrus pulp and molasses (CM). For BM and CM, the diet composition was the same as that of CORN except that a portion of the corn was replaced with barley and molasses or citrus pulp and molasses, respectively, on a dry matter basis. Cows grazed ryegrass (Lolium multiflorum Lam.) pasture. Yield of milk, 3.5% fat-corrected milk, energy-corrected milk, and milk fat, as well as milk fat percentage, were not different among treatments. True milk protein percentage was higher for CORN (2.81%) compared with CM (2.70%), but was not different for BM (2.77%). However, true milk protein yield was not different among treatments. Milk urea N was higher for BM (11.43 mg/dL) compared with both CORN and CM (average: 9.95 mg/dL). There were no differences among CORN, BM, and CM treatments for overall BUN (average: 10.60 mg/dL). At 0400 h, however, cows on CORN had higher BUN than cows on CM (11.43 vs. 9.96 mg/dL), but there were no differences between CORN and BM (average: 11.21 mg/dL) or BM and CM (average: 10.48 mg/dL), and there were no differences among treatments at other time points. The CM diet might have shown more advantage if the pasture crude protein content was higher. Partial replacement of corn with citrus pulp for grazing cows should be further studied using pasture with higher crude protein content. Although cows receiving CM and BM did not produce more milk than cows on CORN, if barley or citrus pulp is less expensive than corn, they may be viable replacements for a portion of the corn supplement for grazing cows.

Nutrient content of whole cottonseed.

The objective of this study was to determine if the nutrient and gossypol contents and in vitro digestibility of 3 types of genetically modified whole cottonseed differed from traditional whole cottonseed. Samples of seed from traditional (no genetic modifications) and genetically modified varieties of cotton grown in 1999 and 2000 were analyzed. Genetic modifications included the insertion of genes to protect cotton from insect pests (Bt), and damage from glyphosate herbicides (RR), and from both (Bt/RR). Year effects were significant for in vitro dry matter (DM) digestibility, gossypol, DM, crude protein (CP), fat, neutral detergent fiber (NDF), acid detergent fiber (ADF), and ash. Higher rainfall resulted in higher CP, fat, and ash and lower NDF and gossypol. There were no differences among seed types for ground or whole seed digestibility, DM, CP, fat, NDF, ADF, ash, lignin, net energy for lactation, amino acids, total fatty acids, or seed index. Overall, the nutrient content and digestibility of varieties of genetically modified seed were similar to that of varieties of traditional whole cottonseed.

Ruminal biohydrogenation in Holstein cows fed soybean fatty acids as amides or calcium salts.

Fatty amides of high oleate fats and calcium salts of palm oil were reported to resist biohydrogenation by ruminal microorganisms. This study was conducted to determine whether converting polyunsaturated fat sources to amides and calcium salts had equal ability to resist biohydrogenation. A total mixed ration consisting of forage and concentrate contained (dry basis): 1) 2.45% soybean oil (SBO), 2) 2.75% calcium salt of SBO, 3) 2.75% amide of SBO, or 4) 2.75% of a mixture of the calcium salt and amide (80:20, wt/wt) of SBO. The 4 diets were fed ad libitum to 4 multiparous lactating Holstein cows fitted with ruminal cannulas in a 4 x 4 Latin square with 21-d periods. Omasal samples were taken to measure postruminal fatty acid content and determine the extent of ruminal biohydrogenation. Adding SBO to the diets as either calcium salts or amides increased omasal flow of C18:2 (n-6) from 25 to 39 g/d. Omasal flow of C18:1 increased from 36 to 49 g/d when SBO was fed to cows as calcium salts, but increased to 86 g/d when SBO was fed as amides. Adding the soybean amide to the diet more than doubled the delivery of C18:1 (n-9) to the omasum of lactating cows, but it also increased trans fatty acid production in the rumen accompanied by milk fat depression. In this study, calcium salts and amide derivatives of fatty acids were both effective in enhancing omasal flow of unsaturataed fatty acids in lactating dairy cows. Amides were more effective than calcium salts for increasing the postruminal flow of oleic acid.

Structural characterization of the GSK-3beta active site using selective and non-selective ATP-mimetic inhibitors.

GSK-3beta is a regulatory serine/threonine kinase with a plethora of cellular targets. Consequently, selective small molecule inhibitors of GSK-3beta may have a variety of therapeutic uses including the treatment of neurodegenerative diseases, type II diabetes and cancer. In order to characterize the active site of GSK-3beta, we determined crystal structures of unphosphorylated GSK-3beta in complex with selective and non-selective ATP-mimetic inhibitors. Analysis of the inhibitors' interactions with GSK-3beta in the structures reveals how the enzyme can accommodate a number of diverse molecular scaffolds. In addition, a conserved water molecule near Thr138 is identified that can serve a functional role in inhibitor binding. Finally, a comparison of the interactions made by selective and non-selective inhibitors highlights residues on the edge of the ATP binding-site that can be used to obtain inhibitor selectivity. Information gained from these structures provides a promising route for the design of second-generation GSK-3beta inhibitors.

Effects of feeding time on nitrogen capture by lactating dairy cows grazing rye pasture.

The objective of this experiment was to determine whether varying times at which a partial mixed ration was fed, either before or after grazing, affected N utilization from rye pasture and thus affected milk yield and components. Sixteen Holstein cows were fed a partial mixed ration (PMR) either at 0700, 0830, or 1100 h. Cows were milked at 0900 h and turned out to graze at 0930 h. Treatments represented feeding times 2.5 h and 1 h before grazing and immediately after grazing. The study was conducted as a 3 x 3 Latin square with three 17-d periods. There were no significant differences among treatments for pasture intake or yield of milk or milk components. Milk yield, fat %, and protein % were 29.4, 29.6, and 29.3 kg, 3.5, 3.5, and 3.4%, and 3.4, 3.5, and 3.4% for treatments, respectively. The milk urea levels were 15.6, 15.1, and 15.5 mg/dl, and were not different among treatments. Blood samples were collected on the last day of each period at 0645, 0845, 1045, 1200, and 1400 h. Blood urea nitrogen (BUN) was measured as an indicator of ruminal N capture. Concentrations were not significantly different among diets before grazing; however, they were significantly different among all treatments approximately 1 h after cows were removed from pasture. Cows fed at 0700 h, 2 h before grazing, maintained lower BUN levels across the 7 h during which the blood samples were collected. Cows that ate the PMR immediately after grazing maintained the highest BUN. Feeding a PMR to cows that graze at different times before and after grazing affected the capture of ruminal N, as indicated by differences in the levels of BUN, but there was no effect on yield of milk or milk components.

Comparison of fatty acid content of milk from Jersey and Holstein cows consuming pasture or a total mixed ration.

Holstein (n = 19) and Jersey (n = 18) cows were used to study effects of two feeding systems on fatty acid composition of milk. Confinement cows were fed a total mixed ration with corn silage and alfalfa silage and pastured cows grazed a crabgrass (90%) and clover (10%) pasture and were allowed 5.5 kg of grain per head daily. Two milk samples were collected from each cow at morning and afternoon milkings 1 d each week for four consecutive weeks in June and July 1998. One set of milk samples was analyzed to determine fatty acid composition, and the second set was used for crude protein and total fat analyses. Data were analyzed by the general linear models procedure of SAS, using a split-plot model with breed, treatment, and breed x treatment as main effects and time of sampling and week as subplot effects along with appropriate interactions. Milk from pastured cows was higher than milk from confinement cows for the cis-9, trans-11 octadecadienoic acid isomer of conjugated linoleic acid (CLA). Also, milk from Holsteins was higher than milk from Jerseys for C16:1, C18:1, and CLA and lower than Jerseys for C6:0, C8:0, C10:0, C12:0, and C14:0. Several treatment x week interactions existed, but main effects were still important; for example, proportions of CLA in milk of grazed cows were relatively constant across weeks (0.66, 0.64, 0.64, and 0.69% +/- 0.02%, respectively), but the CLA in milk of confinement cows increased in wk 4 (0.35, 0.31, 0.31, and 0.48% +/- 0.02% for wk 1 to 4, respectively). There are potentially important differences in fatty acid composition of milk from cows consuming a warm season pasture species compared with milk from cows consuming a total mixed ration, as well as differences between Holstein and Jersey breeds.

"Open" structures of MurD: domain movements and structural similarities with folylpolyglutamate synthetase.

UDP-N-acetylmuramoyl-l-alanine:d-glutamate (MurD) ligase catalyses the addition of d-glutamate to the nucleotide precursor UDP-N-acetylmuramoyl-l-alanine (UMA). The crystal structures of Escherichia coli in the substrate-free form and MurD complexed with UMA have been determined at 2.4 A and 1.88 A resolution, respectively. The MurD structure comprises three domains each of a topology reminiscent of nucleotide-binding folds. In the two structures the C-terminal domain undergoes a large rigid-body rotation away from the N-terminal and central domains. These two "open" structures were compared with the four published "closed" structures of MurD. In addition the comparison reveals which regions are affected by the binding of UMA, ATP and d-Glu. Also we compare and discuss two structurally characterized enzymes which belong to the same ligase superfamily: MurD and folylpolyglutamate synthetase (FGS). The analysis allows the identification of key residues involved in the reaction mechanism of FGS. The determination of the two "open" conformation structures represents a new step towards the complete elucidation of the enzymatic mechanism of the MurD ligase.

In situ disappearance of malate from alfalfa and bermudagrass hay.

The objectives of this study were to determine the rate of malate and dry matter disappearance from different forages in the rumen. Four nonlactating, ruminally cannulated Holstein cows were fed a hay-based diet. Samples of early and late harvested alfalfa, Coastal bermudagrass, and Tifton 85 bermudagrass hays were ground, placed in nylon in situ bags, and ruminally incubated for 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 h. After incubation, samples were rinsed, freeze-dried, extracted, and analyzed for malate content by HPLC with an organic acid column. When forages were incubated in the rumen, malate concentrations were less than 0.55 mg/g of dry matter at 0.5 h and remained low for the 48-h incubation period. These results suggest that malate was solublized and utilized within 30 min after reaching the rumen. Dry matter digestibility of both forages increased with time and was different across forages. Both alfalfa samples were digested to a greater extent between 0.5 and 24 h than either type of bermudagrass, but after 48 h the early maturity Tifton 85 digestibility was similar to alfalfa. Even though it is more common to feed unground forages to ruminants, these in situ results suggest that once malate is available in the rumen it will disappear quickly.

Mechanism of calcite crystal growth inhibition by the N-terminal undecapeptide of lithostathine.

Pancreatic juice is supersaturated with calcium carbonate. Calcite crystals therefore may occur, obstruct pancreatic ducts, and finally cause a lithiasis. Human lithostathine, a protein synthesized by the pancreas, inhibits the growth of calcite crystals by inducing a habit modification: the rhombohedral (10 14) usual habit is transformed into a needle-like habit through the (11 0) crystal form. A similar observation was made with the N-terminal undecapeptide (pE(1)R(11)) of lithostathine. We therefore aimed at discovering how peptides inhibit calcium salt crystal growth. We solved the complete x-ray structure of lithostathine, including the flexible N-terminal domain, at 1.3 A. Docking studies of pE(1)R(11) with the (10 14) and (11 0) faces through molecular dynamics simulation resulted in three successive steps. First, the undecapeptide progressively unfolded as it approached the calcite surface. Second, mobile lateral chains of amino acids made hydrogen bonds with the calcite surface. Last, electrostatic bonds between calcium ions and peptide bonds stabilized and anchored pE(1)R(11) on the crystal surface. pE(1)R(11)-calcite interaction was stronger with the (11 0) face than with the (10 14) face, confirming earlier experimental observations. Energy contributions showed that the peptide backbone governed the binding more than did the lateral chains. The ability of peptides to inhibit crystal growth is therefore essentially based on backbone flexibility.

Role of the ortholog and paralog amino acid invariants in the active site of the UDP-MurNAc-L-alanine:D-glutamate ligase (MurD).

To evaluate their role in the active site of the UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase (MurD) from Escherichia coli, 12 residues conserved either in the Mur superfamily [Eveland, S. S., Pompliano, D. L., and Anderson, M. S. (1997) Biochemistry 36, 6223-6229; Bouhss, A., Mengin-Lecreulx, D., Blanot, D., van Heijenoort, J., and Parquet, C. (1997) Biochemistry 36, 11556-11563] or in the sequences of 26 MurD orthologs were submitted to site-directed mutagenesis. All these residues lay within the cleft of the active site of MurD as defined by its 3D structure [Bertrand, J. A., Auger, D., Fanchon, E., Martin, L., Blanot, D., van Heijenoort, J., and Dideberg, O. (1997) EMBO J. 16, 3416-3425]. Fourteen mutant proteins (D35A, K115A, E157A/K, H183A, Y194F, K198A/F, N268A, N271A, H301A, R302A, D317A, and R425A) containing a C-terminal (His)(6) extension were prepared and their steady-state kinetic parameters determined. All had a reduced enzymatic activity, which in many cases was very low, but no mutation led to a total loss of activity. Examination of the specificity constants k(cat)/K(m) for the three MurD substrates indicated that most mutations affected both the binding of one substrate and the catalytic process. These kinetic results correlated with the assigned function of the residues based on the X-ray structures.

Interaction of tallow and hay particle size on ruminal parameters.

Four nonlactating ruminally cannulated Holstein cows were used in a 4 x 4 Latin square experiment with 4 21-d periods to determine if the effects of dietary fat would be affected by hay particle length. Treatments consisted of two levels of tallow (0 and 5%) and two hay particle lengths (short-cut and long-cut) in a 2 x 2 factorial. Diets contained alfalfa hay, corn silage, and concentrate [1:1:2, dry matter (DM) basis] fed as a total mixed ration (TMR) once per day. Samples of the 0 and 5% tallow TMR were ground and incubated in situ in polyester bags for 24 and 48 h. Ruminal samples were taken on day 21 at 0800 h and at 2-h intervals until 1600 h. The total tract digestibilities of acid detergent fiber (ADF) and neutral detergent fiber (NDF) were not affected by tallow or by hay by tallow interactions. There was a trend for tallow to improve total tract digestibility of crude protein (CP) (70.2 vs. 74.7%). After 48 h of ruminal incubation, tallow significantly decreased the digestibilities of DM, ADF, and NDF. No hay length by tallow interactions for DM, NDF, ADF or CP digestibilities occurred after 24 or 48 h. Tallow increased concentrations of propionate and decreased concentrations of acetate and valerate and the acetate-to-propionate ratio. Total volatile fatty acids increased when tallow was added to diets with short-cut hay, which suggests that when unprotected fat is added to diets with a high level of hay, a short-cut hay length may be advantageous. This result may be due to shorter rumen retention time of feed particles, which reduces the time for fatty acids to exert antimicrobial effects. Or, it may because the increased surface area of the hay particle provides more area for microbial attachment and increased fermentation.