Using alfalfa leaf meal as a supplement in late-gestation beef heifer and nursing beef calf diets.

Two experiments were conducted to investigate using alfalfa leaf meal (ALM; 22% CP, DM basis) in beef cattle diets. In Exp. 1, a total of 24 late-gestation Angus heifers (initial BW 470 +/- 9 kg) were blocked by BW, calving date, and BCS to 1 of 4 dietary treatments in a randomized complete block design. All heifers were offered a basal hay diet (7.4% CP and 67.6% NDF, DM basis). Treatments were arranged as a 2 x 2 factorial consisting of CP supplied at 100 or 112.5% of the recommended daily intake using either soybean meal (SBM) or ALM as the supplemental protein source. Treatments were fed for an average of 100 d before calving. Total DMI was unaffected by supplemental protein source, although heifers consumed more (P < 0.001) ALM supplement than SBM supplement at the expense of hay and corn. Feeding 112.5% of recommended CP to heifers increased precalving rate of BW gain (P = 0.004) and DM digestibility (P = 0.003). Protein source did not affect DM digestibility (P = 0.17). Neither supplemental protein source nor protein amount affected changes in BCS or calving traits. In Exp. 2, replicates of treatments were conducted over 2 consecutive years at 2 locations in northern Minnesota to determine the effects of including ALM in creep-fed supplements on nursing calf performance, supplement BW gain efficiency (GF; BW gain over control/supplement intake), and cow performance. Treatments were control (no supplement), ALM supplement (58% ALM, as-fed basis), or a wheat middling- and soybean hull-based supplement (MIDD). Milk intake (estimated by the weigh-suckle-weigh technique) was similar among treatments. Creep-fed calves had greater (P < 0.001) ADG than control calves, whereas calves offered MIDD tended to have greater ADG (P = 0.05) than those offered ALM (1.38 vs. 1.30 kg/d, respectively). Calves offered MIDD had greater (P < 0.001) creep feed DMI than those offered ALM (2.6 vs. 1.3 kg/d, respectively). A year x treatment interaction was noted for GF (P = 0.02). In yr 1, GF for calves offered ALM was greater (P = 0.006) than GF for calves offered MIDD, but in yr 2, there were no differences. Alfalfa leaf meal may substitute for SBM in beef heifer wintering diets and conventional creep feed ingredients. When included in creep feed diets, ALM can result in slightly less ADG and less DMI, but supplement conversion efficiency may be increased.

SEPT9 gene sequencing analysis reveals recurrent mutations in hereditary neuralgic amyotrophy.

BACKGROUND: Hereditary neuralgic amyotrophy (HNA) is an autosomal dominant disorder that manifests as recurrent, episodic, painful brachial neuropathies. A gene for HNA maps to chromosome 17q25.3 where mutations in SEPT9, encoding the septin-9 protein, have been identified. OBJECTIVE: To determine the frequency and type of mutations in the SEPT9 gene in a new cohort of 42 unrelated HNA pedigrees. METHODS: DNA sequencing of all exons and intron-exon boundaries for SEPT9 was carried out in an affected individual in each pedigree from our HNA cohort. Genotyping using microsatellite markers spanning the SEPT9 gene was also used to identify pedigrees with a previously reported founder haplotype. RESULTS: Two missense mutations were found: c.262C>T (p.Arg88Trp) in seven HNA pedigrees and c.278C>T (p.Ser93Phe) in one HNA pedigree. Sequencing of other known exons in SEPT9 detected no additional disease-associated mutations. A founder haplotype, without defined mutations in SEPT9, was present in seven pedigrees. CONCLUSIONS: We provide further evidence that mutation of the SEPT9 gene is the molecular basis of some cases of hereditary neuralgic amyotrophy (HNA). DNA sequencing of SEPT9 demonstrates a restricted set of mutations in this cohort of HNA pedigrees. Nonetheless, sequence analysis will have an important role in mutation detection in HNA. Additional techniques will be required to find SEPT9 mutations in an HNA founder haplotype and other pedigrees.

Transgenic mice expressing a human mutant beta1 thyroid receptor are hyperactive, impulsive, and inattentive.

Attention deficit hyperactivity disorder (ADHD) is the most commonly diagnosed childhood psychiatric disorder. We have found that a transgenic mouse bearing a human mutant thyroid receptor (TRbeta1) expresses all of the defining symptoms of ADHD--inattention, hyperactivity, and impulsivity--as well as a 'paradoxical' response to methylphenidate (MPH). As with ADHD, the behavioral phenotypes expressed by the TRbeta transgenic mice are dynamic and sensitive to changes in environmental conditions, stress, and reinforcement. TRbeta transgenic mice are euthyroid except for a brief period during postnatal development, but the behavioral phenotypes, elevated dopamine turnover, and paradoxical response to MPH persist into adulthood. Thus, like the vast majority of children with ADHD, the TRbeta transgenic mice exhibit the symptoms of ADHD in the complete absence of thyroid abnormalities. This suggests that even transient perturbations in developmental thyroid homeostasis can have long-lasting behavioral and cognitive consequences, including producing the full spectrum of symptoms of ADHD.

Fixed-time insemination in peripuberal, lightweight replacement beef heifers after estrus synchronization with PGF2alpha and GnRH.

Estrus synchronization contributes to optimizing the use of time, labor, and financial resources by shortening the calving season, in addition to increasing the uniformity of the calf crop. We determined whether acceptable pregnancy rates could be achieved after synchronization of ovulation and fixed-time artificial insemination (AI) in peripuberal replacement beef heifers using gonadotropin-releasing hormone (GnRH) and PGF2alpha. Crossbred heifers from two herds (MH, n=239; SS, n=330) were wintered at a single location. After a prebreeding examination revealed that 55 heifers had a reproductive tract score (RTS) of 1 (infantile reproductive tracts), they were culled and the remaining heifers were assigned randomly to one of three treatment groups: administration of 25mg PGF2alpha i.m. on Days -12 and 0 followed by estrus detection and insemination between 10 and 14 h after an observed estrus (Control; n=173); administration of 100 microg GnRH i.m. on Day -6, followed by 25 mg PGF2alpha i.m. on Day 0, then fixed-time AI and administration of 100 microg GnRH i.m. on Day +2 (GPG; n=172); and, treatment as for group GPG in addition to administration of 100 microg GnRH i.m. on Day -12 (GGPG; n=169). Bulls were introduced 10 days after AI for 60 days to breed heifers which did not conceive after AI (clean-up bulls). On Days -12, -6, and 0 transrectal ultrasonography was used to monitor ovarian structures in a subset of heifers (30 per treatment). At 30-35 days after AI, ultrasound was used to determine the presence of a viable fetus. Presence of a fetus and stage of pregnancy were determined via palpation per rectum 61-63 days after the conclusion of the breeding season. Heifers in the MH herd (309+/-1.9 kg) were heavier (P<0.001) than those in the SS herd (283+/-1.7 kg) at initiation of the breeding season. Synchronized pregnancy rates were greater (P<0.05) in GGPG (25.4%) and GPG (22.1%) than Control (12.7%) heifers. Pregnancy rates were 9, 21, 32, or 31% for heifers with RTS of 2, 3, 4, or 5, respectively. The average diameter of 22 follicles induced to ovulate in heifers treated with GnRH (GPG and GGPG treatments) was 14.2+/-0.8 mm (range=10.0-23.6 mm). In conclusion, a fixed-time ovulation synchronization program using GnRH and PGF2alpha improved pregnancy rates in peripuberal, lightweight replacement beef heifers.

Reflection on the history, coordination, and funding trends for U.S. public meat research: information to enhance resource allocation.

A study was conducted to analyze resource allocation for public meat research in the United States and characterize the portfolio of meat research investments. Trends in the amount of public resources provided for meat research (beef, pork, lamb, and poultry) were analyzed for fiscal years 1980, 1985, 1990, 1995, and 1997. An in-depth analysis was conducted for data from fiscal year 1998 to characterize the profile of the research portfolio. Funding levels and scientist-year equivalents were aggregated to represent the measures of resource allocation for three mutually exclusive research categories: 1) meat quality, 2) food safety, and 3) product development and processing. Data for the 1998 profile analysis were derived from a computer search based on the combination of key words and research classification codes to avoid duplication and cluster research projects. Individual research projects were individually reviewed and a percentage was assigned to four mutually exclusive research categories: 1) meat quality, 2) food safety, 3) product development and processing, and 4) marketing. As meat research evolved over the past century, considerable efforts were expended by researchers and administrators to ensure the coordination of research and program relevance. This is demonstrated by the establishment of numerous multistate research committees. Total funding for meat science increased only modestly when adjusted for inflation during the two decades of this study; however, notable changes occurred in the distribution of resources in the portfolio. Funding for meat quality and product development and processing remained virtually unchanged when adjusted for inflation, whereas funding for food safety increased considerably. The total number of scientists conducting meat research remained virtually unchanged during the period, but the proportion allocated to food safety research increased substantially. The federal portion of total funding decreased from 61.3% to 51.6% between 1980 and 1997, whereas the percentage from both state appropriations and private sources increased. Modifications in research emphasis were influenced by industry problems such as meat quality, public perceptions about food safety, the availability of research funding, scientific advances occurring in molecular biology and genetic manipulation, and the changing meat industry. The information in this paper provides administrators and researchers the opportunity to make better informed decisions about resource allocation for meat research.

Phylogenetic analysis and trait evolution in Australian lineages of drywood termites (Isoptera, Kalotermitidae).

A phylogenetic analysis of Australian drywood termites (Isoptera, Kalotermitidae) based on partial sequence from the cytochrome oxidase II (COII) and cytochrome b genes is presented. In addition to providing new information on the evolutionary relationships among 25 species from seven genera, we evaluate the relative likelihoods of alternative topological hypotheses, including those derived from morphology-based classifications. We also test the applicability of a molecular clock for estimating the age of the Kalotermitidae and infer the evolution of species-specific variation for habitat type and soldier caste phragmosis by mapping this information onto the independently derived phylogeny. Maximum-likelihood analysis of both nucleotide and protein sequences from a multigene data set jointly support a single topology, which is shown to be the best estimate of the true phylogeny among the alternatives tested. Our results support the monophyly of all genera but question the discrimination between Procryptotermes and Cryptotermes. A basal dichotomy among generic groups suggests two principle lines of divergence within the family. Intergeneric relationships show mixed congruence to previous proposals, resulting in one morphology-based classification being rejected. A molecular clock hypothesis is not supported due to significant among-lineage rate heterogeneity in the COII gene. Patterns revealed through trait mapping suggest that the most recently diverged taxa tend to occupy the driest habitats and that these same taxa reflect a defensive transition away from large mandibulate soldiers toward small phragmotic soldiers. The association between habitat and defensibility supports the hypothesis that these two characters have been tightly linked throughout the social diversification of termites.

Cause of metabolic acidosis in prolonged surgery.

OBJECTIVE: The intraoperative development of metabolic acidosis is frequently attributed to hypovolemia, tissue hypoperfusion, and lactic acidosis. In this study, dilutional acidosis was evaluated as a possible mechanism for the routine development of intraoperative acidosis in noncardiac, nonvascular surgery patients. DESIGN: Prospective, observational study. SETTING: University-affiliated Veteran's Affairs Medical Center and a staff model, health maintenance organization hospital. PATIENTS: Twelve patients undergoing prolonged surgical procedures expected to last > or = 4 hrs were enrolled in the study. INTERVENTIONS: Perioperative management was based on the judgment of the attending anesthesiologist and surgeon without knowledge of the study's intent. MEASUREMENTS AND MAIN RESULTS: Arterial blood gas parameters, serum electrolytes, and urine electrolytes were measured pre- and postoperatively. Pulmonary artery catheters were placed for hemodynamic measurement and oxygen delivery calculations. Plasma volume was measured both pre- and postoperatively, using the Evans blue dye dilution technique. Although significant changes in lactate level (1.1 +/- 0.6-1.8 +/- 1.0) occurred, the change was not large enough to explain the degree of change in base excess (0.8 +/- 2.3 to -2.7 +/- 2.9). Chloride levels significantly increased (106 +/- 3-110 +/- 5) with a correlation (r2 = .92; p < .0001) between the degree of change in chloride and the degree of change in base excess. Plasma volume did not change. Furthermore, a correlation between the volume of normal saline administered and the change in base excess was found (r2 = .86; p < .0001), although no correlation was found with Ringer's lactate solution. An even stronger correlation was noted when the total chloride amount administered was compared with the change in base excess (r2 = .93; p < .0001). CONCLUSIONS: In this patient population, a common source of increasing base deficit is related to chloride administration. The largest source of chloride is usually normal saline. Classically, dilutional acidosis would explain the predominance of this acidotic change; however, no increase in plasma volume occurred. The absence of plasma volume change would suggest that the mechanism postulated to result in dilutional acidosis is incomplete. The common treatment of administering more fluid for intraoperative acidosis may be inappropriate, may have caused the acidosis, and may further exacerbate the acidosis. Chloride levels should be assessed whenever a metabolic acidosis is seen perioperatively.

Influence of radical prostatectomy on serum hormone levels.

PURPOSE: The influence of radical prostatectomy on the hypothalamic pituitary axis has not been well studied. It is also unclear how alterations in serum androgen levels that result from surgical removal of the prostate might influence the recovery of libido and sexual function following radical prostatectomy. We determined the influence of radical prostatectomy on the hypothalamic pituitary testicular axis of 63 men with clinically localized prostate cancer treated only with radical prostatectomy. MATERIALS AND METHODS: A total of 63 healthy men 43 to 67 years old were enrolled in this prospective study. Phlebotomy was performed immediately before and 1 year following radical retropubic prostatectomy. Sera were stored frozen and analyzed as a group at the end of the study. We measured serum testosterone, percent free testosterone, dihydrotestosterone (DHT), estradiol, luteinizing hormone (LH), follicle-stimulating hormone (FSH), sex hormone binding globulin and prolactin. RESULTS: Following radical prostatectomy there was a statistically significant increase in serum testosterone, free testosterone, estradiol, LH and FSH (p <0.0001), and statistically significant decrease in serum DHT (p <0.0001). No difference was noted in serum sex hormone binding globulin or prolactin levels. There was no statistically significant correlation between any serum hormone and sample storage time, patient age or prostate volume that could limit potential bias in study design. Serum hormone changes did not correlate with pathological stage or histological grade for this group of patients. CONCLUSIONS: Radical prostatectomy influences the hypothalamic pituitary axis by increasing serum testosterone, percent free testosterone, estradiol, LH and FSH while decreasing serum DHT levels. These findings suggest that the sexual dysfunction associated with radical prostatectomy cannot be explained by androgen deficiency alone. These data further suggest that the normal prostate and/or prostate neoplasm could secrete a substance or substances that give negative feedback control to pituitary gonadotropin secretion. Further investigation is warranted to identify this substance or substances.

RoBo-1, a novel member of the urokinase plasminogen activator receptor/CD59/Ly-6/snake toxin family selectively expressed in rat bone and growth plate cartilage.

Using differential display polymerase chain reaction, we cloned a novel cDNA named RoBo-1 from rat tibia. RoBo-1 is abundantly expressed in bone, including the hypertrophic chondrocytes of the growth plate where cartilage is remodeled into bone. RoBo-1 mRNA expression increased in response to two modulators of bone metabolism, estradiol and intermittent mechanical loading, suggesting a role in bone homeostasis. The 1.6-kilobase cDNA encodes a 240-amino acid protein with a cysteine spacing pattern, suggesting that RoBo-1 is a novel member of the urokinase plasminogen activator receptor/CD59/Ly-6/snake toxin family. Furthermore, the C-terminal contains a glycosyl-phosphatidylinositol attachment site, suggesting that it is a cell surface protein similar to other mammalian members of this family. The strongest homology of RoBo-1 is to the snake serum-derived phospholipase A2 inhibitors, which uniquely contain two of the cysteine domains but are secreted proteins. Interestingly, RoBo-1 is likely the first membrane-anchored member of this family containing two cysteine domains. Thus, the tissue specificity, responsiveness to bone protective mediators, along with its relationship to the multifunctional urokinase plasminogen activator receptor/CD59/Ly-6/snake toxin family suggests that RoBo-1 may play a novel role in the growth or remodeling of bone.

Cyclic AMP signaling pathways are important in IL-1 beta transcriptional regulation.

An intact cAMP response element (CRE) in the upstream regulatory sequence of IL-1 beta (-2755/-2762) has been shown to be essential for maintaining full IL-1 beta inducibility following treatment with LPS, PMA, or TNF-alpha. In the present study, using the recombinant plasmid pIL-1(4.0 kb)-chloramphenicol acetyltransferase, containing 4.0 kb of the IL-1 beta upstream regulatory sequence, we have demonstrated that dibutyryl cAMP treatment alone is capable of induction. Due to the critical nature of the CRE for the induction of IL-1 beta transcription, an effort was made to determine the importance of the cAMP signaling pathway(s) by determining whether CRE binding protein (CREB) and other CREB/activating transcription factor (ATF) family members that responded to cAMP were associated with the DNA-protein complex that forms at this site. Nuclear extracts prepared from LPS-treated THP-1 5A cells were fractionated by ammonium sulfate precipitation and heparin-Sepharose chromatography, and the resulting fractions were characterized in electrophoretic gel mobility shift assays. These purification steps resulted in an approximately 100-fold enrichment of the proteins binding to the CRE site. Western blot analysis of isolated fractions, using CREB- and ATF-1-specific Ab showed an increased level of these proteins in the enriched fractions. Tryptic digest and DNase I protection studies showed the presence of CREB protein in the complex at the CRE site. Supershift electrophoretic gel mobility shift assays and immunoprecipitation analysis provided further evidence that both CREB and ATF-1 are present in the complex. In addition, an increase in CREB phosphorylation was observed when THP-1 5A cells were treated with dibutyryl cAMP, LPS, or both. These studies confirm the importance of a cAMP signaling pathway(s) in the regulation of IL-1 beta at the transcriptional level.

MR imaging of breast implants.

Magnetic resonance (MR) images of 57 implants in 32 women were reviewed for possible complications. Spin-echo T2-weighted MR imaging of one or both breasts was performed with dedicated breast coils in axial, coronal, and sagittal planes. At surgery in 19 patients, MR evaluation for rupture correlated in 17 cases (two false-negative ruptures). Implant ruptures were seen at surgery in 15 of the 19 patients. Nine ruptures involved single-lumen implants: Six were ruptured within the fibrous capsule that normally forms around the foreign implant (intracapsular rupture [two were seen only at surgery]), and three were ruptured beyond this fibrous capsule into the soft tissues (extracapsular rupture). Five ruptures involved double-lumen implants, in which only the outer lumen had ruptured, and one involved a single-lumen saline implant that had completely collapsed. Additional complications of capsule formation and infection were suggested in two implants, and infection was suggested in one. The MR imaging appearances of the various types of implants as well as their complications are presented.

Evaluation of immunoperoxidase techniques to detect Rickettsia rickettsii in fixed tissue sections.

Standard immunoperoxidase techniques were evaluated in the diagnosis of Rocky Mountain spotted fever (RMSF). Formalin-fixed, paraffin-embedded tissue was tested to detect Rickettsia rickettsii using the same antibody provided by the Centers for Disease Control and Prevention that is used for direct immunofluorescence (DIF). Tissues from 23 patients with suspected RMSF were divided: some were snap-frozen for DIF; the remainder were fixed in formalin, processed routinely, and embedded in paraffin for immunoperoxidase and hematoxylin and eosin staining. Ten patients were ultimately determined to have RMSF; in nine of these patients, both DIF and immunoperoxidase staining were positive for R rickettsii. There were no discrepancies, positive or negative, between the two methods. In the one case with a false-negative result by both methods, the patient had received antirickettsial antibiotics 72 hours before the biopsy was performed. These data demonstrate that the immunoperoxidase technique is effective in diagnosing RMSF and may be applied to cases retrospectively.

An immunohistochemical study of perianal Paget's disease. Possible origins and clinical implications.

The histogenesis of perianal Paget's disease is controversial. A clinical and pathologic study was done of a patient with a history of adenocarcinoma of the rectum for whom a subsequent diagnosis of perianal Paget's disease was the sole manifestation of recurrent rectal cancer. Immunohistochemical techniques were used to compare and contrast the original rectal adenocarcinoma with the subsequent perianal skin recurrence confined to the epidermis. Both the rectal adenocarcinoma and the Paget's cells were positive for cytokeratin, epithelial membrane antigen, B72.3, and carcinoembryonic antigen and negative for gross cystic disease fluid protein-15, Leu-M1, CA 125, and S-100 protein. These findings, their relevance to the histogenesis of perianal Paget's disease, and the possible clinical implications are discussed.

Improved lung preservation with cold air storage.

Conventional topical slush cooling limits lung transport to 4 to 6 hours. For this canine study of an alternate air cooling system, 37 canine lungs were removed: 24 were placed in plastic bags, and inserted in a Transplanthermm container at core air temperatures (n = 6 lungs each) of (A) 4 degrees C, (B) 8 degrees C, (C) 12 degrees C, and (D) 20 degrees C; 6 were stored conventionally in ice slush (E); and 7 were transplanted immediately (F). After 8 hours, the stored lungs were transplanted and the contralateral pulmonary artery was ligated. Survival, arterial oxygen tension, and extravascular lung water were monitored at 15 minutes and every hour for 4 hours. Four-hour survival was 100% in groups A, B, and F; 83% in group C, 50% in group D, and 17% in group E. The mean arterial oxygen tension at 1 hour was lower in group E (6.4 +/- 2.4 kPa) than in group A (39.8 +/- 13.2 kPa) (p = 0.0002) or in group F (42.0 +/- 16.2 kPa) (p = 0.0035). Extravascular lung water in group E was higher at 15 minutes (15.44 +/- 5.63 mL/kg) than in group A (3.76 +/- 0.63 mL/kg) (p = 0.0001) and group F (4.69 +/- 1.65 mL/kg) (p = 0.003). Cold air storage appears to provide better lung preservation than hypothermic immersion in ice slush.

Effect of desmopressin on vein graft patency in a microvascular model.

Desmopressin acetate decreases blood loss after cardiac surgery by activating platelets. We studied whether this effect was detrimental to small-caliber vein grafts in rats. Thirty minutes before femoral artery grafting with 0.75-mm-diameter reverse autogenous saphenous vein grafts, 20 rats received desmopressin acetate intravenously at 1.0 micrograms/kg over 10 minutes, and 20 control rats received normal saline intravenously over 10 minutes. In each group, 10 rats received a 6-mm-long graft and 10 an 18-mm-long graft. Graft patency was evaluated at 20 minutes, 24 hours, and 30 days. Intimal thickening was assessed by light and scanning electron microscopy. At 30 days, 9 short grafts and 8 long grafts in the desmopressin-treated group were patent, whereas only 8 short control grafts and only 6 long control grafts were patent. Intimal thickening and platelet deposition were the same in both groups. These data show no detrimental effects of desmopressin acetate on saphenous vein graft patency.