Effects of increased levels of supplemental vitamins during the summer in a commercial artificial insemination boar stud.

Heat stress due to increasing extremes in ambient temperature and humidity results in reduced semen quality in boars. This has caused reduced efficiency of the swine industry, requiring more boars to breed the same number of sows. Vitamins such as vitamin C (VC) and E (VE) have been shown to improve semen quality in boars. Recently, vitamin D has been shown to improve semen quality in boars. The purpose of this experiment was to evaluate the effects of increased supplemental vitamins on boar reproduction during the summer season in a commercial boar stud. One hundred and sixty Pig Improvement Company (PIC) terminal line boars (n = 32 per treatment) and 39 maternal, heat-sensitive boars (n = 7 or 8 per treatment) were randomly allocated to treatment and fed a corn and soybean meal-based diet adjusted based on individual boar body condition score. A control (CNT) diet was used that met PIC recommendations for boars. Increased supplementation of specific vitamins was given in the form of a top-dress and consisted of CNT wheat middlings, CNT plus VC (560 mg/day), CNT plus 25-hydroxy vitamin D3 (VD) (125 µg/day), CNT plus VE (275 mg/day) and CNT plus VC, VD and VE (CDE). The experiment was split into three periods based on maximum daily high temperatures in the barn, where period 1 was weeks 1 to 4, period 2 was weeks 5 to 11 and period 3 was weeks 12 to 14. Semen was collected from boars as needed using the stud's normal production schedule and was analyzed for sperm quantity and quality characteristics. There were no dietary effects on semen volume, sperm concentration or total sperm production (P ≥ 0.553). Total motility of sperm was not impacted by diet (P = 0.115); although, VC tended (P = 0.064) to have a greater progressive motility than CDE. Percentages of morphologically normal sperm and normal acrosomes were not affected by dietary supplementation (P ≥ 0.157). Period effects were observed for most semen quality parameters, with quality generally becoming reduced over time. The present study demonstrates that increased supplementation of vitamins beyond PIC recommendations was not beneficial for boar reproduction during the summer.

Left renal vein ligation: a technique to mitigate low portal flow from splenic vein siphon during liver transplantation.

Low portal vein flows in liver transplant have been associated with poor allograft survival. Identifying and ameliorating causes of inadequate portal flow is paramount. We describe successful reversal of significant splenic vein siphon from a spontaneous splenorenal shunt during liver transplant. The patient is a 43-year-old male with cirrhosis from hepatitis C and Budd-Chiari syndrome, who had a variceal hemorrhage necessitating an emergent splenorenal shunt with 8 mm PTFE graft. Imaging in 2006 revealed thrombosis of the splenorenal shunt and evidence of a new spontaneous splenorenal shunt. The patient developed hepatocellular carcinoma and underwent transplant in 2009. After reperfusion, portal flows were low (150-200 mL/min). A mesenteric varix was ligated without improvement. Due to adhesions, direct collateral ligation was not attempted. In order to redirect the splenic siphon, the left renal vein was stapled at its confluence with the inferior vena cava. Portal flows subsequently increased to 1.28 L/min. Postoperatively, the patient had stable renal and liver function. We conclude that spontaneous splenorenal shunts can cause low portal flows. A diligent search for shunts with understanding of flow patterns is critical; ligation or rerouting of splanchnic flow may be necessary to improve portal flows and allograft outcomes.

Biliary complications after orthotopic liver transplantation from donors after cardiac death: broad spectrum of disease.

BACKGROUND: Donation-after-death liver transplantation (DCD-LT) carries higher complication rates compared with donation-after-brain death liver transplantation (DBD-LT). In this report we describe our experience with biliary complications in DCD-LT with emphasis on anatomical patterns and outcomes. MATERIALS AND METHODS: We performed retrospective review of patients' medical records from August 2004 to December 2008, during which time total of 26 DCD-LTs were performed. Mean follow-up was 29 months (range 3 to 51 months). RESULTS: Biliary complications occurred in 12 patients (46%), of whom 9 were related to DCD (35%). Four patients had more than 1 biliary complication, and 4 had concomitant arterial problems (stricture/thrombosis). Treatment of complications included: ERCP (n = 5, 3 resolved), conversion to roux (n = 5, 2 resolved), revision of roux (n = 1), percutaneous transhepatic cholangiography (n = 1), artery revision (n = 3). Three patients with casts had operative extraction of casts depicting a mummified biliary tree; histology showed casts and fibrosis and anastomotic suture material. Six patients underwent retransplantation (23%). Among retransplanted patients, 2 deaths occurred (7.7%). CONCLUSION: Our experience with DCD-LT reveals a high prevalence of biliary complications with a new and wide spectrum of clinicopathologic findings. Better strategies for prevention of these unique biliary complications are needed to better justify the added risks and costs for performance of DCD-LT.

Improving organ procurement practices in Michigan.

Travel to procure deceased donor organs is associated with risk to transplant personnel. In many instances, multiple teams are present for a given operation. We studied our statewide experience to determine how much excess travel this redundancy entails, and generated alternate models for organ recovery. We reviewed our organ procurement organization's experience with deceased donor operations between 2002 and 2008. Travel was expressed as cumulative person-miles between procurement team origin and donor hospital. A model of minimal travel was created, using thoracic and abdominal teams from the closest in-state center. A second model involved transporting donors to a dedicated procurement facility. Travel distance was recalculated using these models, and mode and cost of travel extrapolated from current practices. In 654 thoracic and 1469 abdominal donors studied, the mean travel for thoracic teams was 1066 person-miles and for abdominal teams was 550 person-miles. The mean distance traveled by thoracic and abdominal organs was 223 miles and 142 miles, respectively. Both hypothetical models showed reductions in team travel and reliance on air transport, with favorable costs and organ transport times compared to historical data. In summary, we found significant inefficiency in current practice, which may be alleviated using new paradigms for donor procurement.

Genetic determinants of response to chemotherapy in transgenic mouse mammary and salivary tumors.

Several transgenic mouse tumor models were utilized to explore how specific genetic alterations affect the tumor cell response to chemotherapeutic agents in vivo. Specifically, MMTV-ras transgenic mice were interbred to p53 knock-out mice to create a model for assessing the role of p53 in chemotherapeutic responses. In addition, MMTV-ras tumors were compared to MMTV-myc and MMTV-ras/myc tumors. Mice of each genotype reproducibly develop mammary and/or salivary tumors, but tumor growth dynamics vary considerably between genotypes. MMTV-ras/p53-/- tumors exhibit higher S phase fractions than MMTV-ras/p53+/+ tumors, although both tumor types display very low apoptosis levels. In contrast, MMTV-myc tumors exhibit both high S phase fractions and spontaneous apoptosis levels. Tumor-bearing mice of each genotype were treated with either doxorubicin or paclitaxel, and effects on overall tumor growth, cell cycle distribution and apoptosis were evaluated. Surprisingly, neither agent efficiently induced apoptosis in any of the tumor models, including those with wildtype p53. Rather, tumor responses were mediated primarily by changes in cell cycle distribution. However, the spontaneous apoptosis levels did serve as a predictor of tumor growth response, in that only those tumors with high pretreatment apoptosis levels underwent significant regression following treatment with either agent.

Reduction of posterior hip dislocations in the lateral position using traction-countertraction: safer for the surgeon?

Closed reduction of a hip dislocation is a physically demanding task for the orthopaedic surgeon. The most commonly used methods for reduction of the hip involve vigorous axial traction on the lower extremity with the patient in the supine position, using an assistant who attempts to hold the pelvis down. The surgeon generally stands over the patient to pull up on the bent knee, which puts the surgeon at risk for a low back injury and, if done while the patient remains on a stretcher, can put the surgeon at risk for a fall from a height. Reduction in the prone position is advocated by some, but caring for the sedated patient in the prone position can be difficult and the patient's pelvis tends to roll off the edge of the stretcher, preventing the achievement of hip flexion, which is desirable in achieving reduction. We describe a traction-countertraction technique that appears to be significantly less dangerous for the surgeon. In this technique the hip is reduced with the patient remaining on the stretcher in the lateral position. In addition, fluoroscopy can be carried out during the reduction maneuver, and the images can be quite helpful in adjusting the direction of manipulative forces.

DRG reimbursement: geriatric hip fractures in the community hospital trauma center.

BACKGROUND: The purpose of this paper was to determine whether Medicare reimbursement for hip fracture reaches cost in geriatric patients. METHODS: We conducted a retrospective review using the hospital trauma registry. Demographics, operations, length of stay, clinical outcome, discharge disposition, hospital charges, and hospital costs were reviewed and compared with diagnosis-related group (DRG) reimbursement. RESULTS: The study included 153 Medicare patients. Mortality was 3.9%, 71% were discharged to a nursing home or rehabilitation unit, and 25% went directly home. DRG reimbursement constituted 58% of charges. Compared with costs, the DRG amount represented a mean loss of nearly $1,000 per patient. CONCLUSIONS: DRG reimbursement undercompensates the community hospital trauma center for treating a common malady among the geriatric population. A population shift toward the elderly, decreasing Medicare remuneration, and the advance of managed care will make correct identification and control of costs extremely important for the hospital caring for hip fractures in the geriatric population.

A farnesyltransferase inhibitor induces tumor regression in transgenic mice harboring multiple oncogenic mutations by mediating alterations in both cell cycle control and apoptosis.

The farnesyltransferase inhibitor L-744,832 selectively blocks the transformed phenotype of cultured cells expressing a mutated H-ras gene and induces dramatic regression of mammary and salivary carcinomas in mouse mammary tumor virus (MMTV)-v-Ha-ras transgenic mice. To better understand how the farnesyltransferase inhibitors might be used in the treatment of human tumors, we have further explored the mechanisms by which L-744,832 induces tumor regression in a variety of transgenic mouse tumor models. We assessed whether L-744,832 induces apoptosis or alterations in cell cycle distribution and found that the tumor regression in MMTV-v-Ha-ras mice could be attributed entirely to elevation of apoptosis levels. In contrast, treatment with doxorubicin, which induces apoptosis in many tumor types, had a minimal effect on apoptosis in these tumors and resulted in a less dramatic tumor response. To determine whether functional p53 is required for L-744,832-induced apoptosis and the resultant tumor regression, MMTV-v-Ha-ras mice were interbred with p53(-/-) mice. Tumors in ras/p53(-/-) mice treated with L-744,832 regressed as efficiently as MMTV-v-Ha-ras tumors, although this response was found to be mediated by both the induction of apoptosis and an increase in G1 with a corresponding decrease in the S-phase fraction. MMTV-v-Ha-ras mice were also interbred with MMTV-c-myc mice to determine whether ras/myc tumors, which possess high levels of spontaneous apoptosis, have the potential to regress through a further increase in apoptosis levels. The ras/myc tumors were found to respond nearly as efficiently to L-744,832 treatment as the MMTV-v-Ha-ras tumors, although no induction of apoptosis was observed. Rather, the tumor regression in the ras/myc mice was found to be mediated by a large reduction in the S-phase fraction. In contrast, treatment of transgenic mice harboring an activated MMTV-c-neu gene did not result in tumor regression. These results demonstrate that a farnesyltransferase inhibitor can induce regression of v-Ha-ras-bearing tumors by multiple mechanisms, including the activation of a suppressed apoptotic pathway, which is largely p53 independent, or by cell cycle alterations, depending upon the presence of various other oncogenic genetic alterations.

Multifiber renal SNA recordings predict mean arterial blood pressure in unanesthetized rat.

The goal of this analysis was to quantify the relationship between renal sympathetic nerve activity (SNA) and mean arterial blood pressure (MAP). We previously recorded renal SNA and MAP in conscious rats during a stressful behavioral stimulus and during a nonstressful stimulus. We then formulated a set of two linear, first-order differential equations that uses our SNA recordings after a time delay (the input) to predict fluctuations in MAP (the output). Our model has four parameters: 1) the cardiovascular time constant T that characterizes the frequency response function between the effector elements controlled by the sympathetic nerves and the cardiovascular system (1-5 s); 2) the effector time constant Te determined by the coupling between the sympathetic nervous system and the effectors (0.0-0.6 s); 3) the efferent time delay tau e between a change in SNA and a change in MAP (0.4-0.6 s); and 4) a proportionality constant C between fluctuations in SNA and fluctuations in MAP (0.3-3.4 mmHg/nV). The parameters of the model were determined that minimize the residual error between the simulated time series and the actual data time series for a stressful stimulus. Then we tested the ability of the transfer function to predict the MAP response to a nonstressful stimulus. In five of seven rats tested, the model's predictions were good, with mean cross-correlation coefficients for the predicted trials between 0.62 and 0.83. We show that multifiber renal SNA recordings can reliably predict changes in MAP in the unanesthetized rat. Thus the overall sympathetic drive to the cardiovascular system is indexed by renal SNA, although the vasomotor effectors driven by renal SNA control only approximately 20% of the blood cow.

Differential regulation of cell cycle characteristics and apoptosis in MMTV-myc and MMTV-ras mouse mammary tumors.

We have used the MMTV-myc and MMTV-ras transgenic mouse mammary tumor models (T. A. Stewart et al., Cell, 38: 627-637, 1984, and E. Sinn et al., Cell, 49: 465-475, 1987) to evaluate how the c-myc and v-Ha-ras oncogenes influence tumor growth characteristics in vivo. MMTV-myc tumors had much higher levels of spontaneous apoptosis than MMTV-ras tumors, whereas intermediate levels were observed in MMTV-myc/ras tumors. Significant differences in cell cycle characteristics were also observed in tumors from mice of the three genotypes. Tumors from MMTV-myc mice had lower G1 and higher S-phase fractions than MMTV-ras tumors, with intermediate values again observed in the MMTV-myc/ras tumors. Despite these differences, however, tumor growth rates for the different groups were similar. These findings highlight the importance of the balance between cell cycle regulation and cell death in determining the kinetics of tumor growth and indicate that distinct oncogenes can have a profound influence on that balance.

Increased tumor proliferation and genomic instability without decreased apoptosis in MMTV-ras mice deficient in p53.

We have used an in vivo tumor model to evaluate the consequences of p53 tumor suppressor protein deficiency in a tissue-specific context. By breeding MMTV-ras transgenic mice, which are highly susceptible to the development of mammary and salivary tumors, with p53(-/-) mice, we generated three classes of animals which contained the MMTV-ras transgene but differed in their p53 functional status (ras/p53(+/+), ras/p53(+/-), or ras/p53(-/-)). ras/p53(-/-) mice developed tumors more rapidly than animals of the other two genotypes; however, the distribution of tumors was unexpectedly altered. Whereas the most frequently observed tumors in ras/p53(+/+) and ras/p53(+/-) mice were of mammary origin, ras/p53(-/-) mice developed primarily salivary tumors. In addition, the mammary and salivary tumors from ras/p53(-/-) mice consistently exhibited a number of unfavorable characteristics, including higher histologic grades, increased growth rates, and extensive genomic instability and heterogeneity, relative to tumors from ras/p53(+/+) mice. Interestingly, the increased growth rates of ras/p53(-/-) tumors appear to be due to impaired cell cycle regulation rather than decreased apoptosis, suggesting that p53-mediated tumor suppression can occur independent of its role in apoptosis.

First-order differential-delay equation for the baroreflex predicts the 0.4-Hz blood pressure rhythm in rats.

We have described a 0.4-Hz rhythm in renal sympathetic nerve activity (SNA) that is tightly coupled to 0.4-Hz oscillations in blood pressure in the unanesthetized rat. In previous work, the relationship between SNA and fluctuations in mean arterial blood pressure (MAP) was described by a set of two first-order differential equations. We have now modified our earlier model to test the feasibility that the 0.4-Hz rhythm can be explained by the baroreflex without requiring a neural oscillator. In this baroreflex model, a linear feedback term replaces the sympathetic drive to the cardiovascular system. The time delay in the feedback loop is set equal to the time delay on the efferent side, approximately 0.5 s (as determined in the initial model), plus a time delay of 0.2 s on the afferent side for a total time delay of approximately 0.7 s. A stability analysis of this new model yields feedback resonant frequencies close to 0.4 Hz. Because of the time delay in the feedback loop, the proportional gain may not exceed a value on the order of 10 to maintain stability. The addition of a derivative feedback term increases the system's stability for a positive range of derivative gains. We conclude that the known physiological time delay for the sympathetic portion of the baroreflex can account for the observed 0.4-Hz rhythm in rat MAP and that the sensitivity of the baroreceptors to the rate of change in blood pressure, as well as average blood pressure, would enhance the natural stability of the baroreflex.

Characterization of the mouse tartrate-resistant acid phosphatase (TRAP) gene promoter.

Tartrate-resistant acid phosphatase (TRAP) is an iron-binding protein that is highly expressed in osteoclasts. To characterize the regulation of TRAP gene expression, progressive 5' and 3' deletions of a 1.8 kb fragment containing the 5'-flanking sequence were fused to a luciferase reporter gene. Two nonoverlapping regions of this 1.8 kb fragment had promoter activity. The upstream promoter (P1) was located within the region from -881 bp to -463 bp relative to the ATG, while the downstream promoter (P2) was located between -363 bp to -1 bp in a region we have previously shown to be an intron in transcripts originating from the upstream promoter. A putative repressor region for the P2 promoter at -1846 bp to -1240 bp and a putative enhancer region at -962 bp to -881 bp relative to the ATG were identified. PCR analysis of promoter-specific transcription of the TRAP gene in various murine tissues showed that both promoters were active in several tissues. Transferrin-bound iron increased P1 promoter activity 2.5-fold and hemin decreased P1 promoter activity, but neither had any effect on P2 activity. These data show that the transcriptional regulation of the TRAP gene is complex and that iron may play a key role in TRAP gene regulation.

A radiation hybrid map of the BRCA1 region.

A locus on chromosome 17q, designated "BRCA1," has been identified as a predisposition gene for breast cancer. A panel of chromosome 17-specific radiation-reduced somatic cell hybrid clones has been assembled for high-resolution mapping of chromosome 17. A series of 35 markers, known to span the BRCA1 locus, were tested against this hybrid panel by PCR assays. Statistical analysis of these data yields a BRCA1 radiation hybrid map at a density sufficient to initiate YAC cloning and pulsed-field gel electrophoretic mapping of the candidate region. In addition, many of the markers reveal genetic polymorphisms and may be tested in breast cancer families and in loss-of-heterozygosity studies of sporadic breast cancers to better define the BRCA1 gene candidate region.

Cloning and characterization of the 5'-flanking region of the mouse tartrate-resistant acid phosphatase gene.

Little information is available on the molecular mechanisms controlling osteoclastic bone resorption. We used tartrate-resistant acid phosphatase (TRAP) to begin to investigate the regulation of bone resorption at the molecular level. TRAP is expressed at high levels in osteoclasts and may play an important role in the bone resorptive process. Therefore, we isolated the murine TRAP gene from a mouse spleen genomic library and characterized its promoter. A restriction map was generated for the 17 kb TRAP insert. A 2 kb SmaI fragment, containing the 5'-flanking region, was subcloned and the nucleotide sequence determined. Sequence analysis of the SmaI fragment revealed the presence of numerous candidate transcription factor binding sequences, including those for AP1 and H-APF-1. The H-APF-1 site matches the consensus sequence for the IL-6-regulated transcription factor. An intron was identified at -1 to -393 bp relative to the ATG. The presence of an intron was confirmed by PCR analysis of RNA isolated from murine osteoclasts. Primer extension analysis indicated the presence of a transcription initiation site at -552 bp from the ATG. The region from -1846 to 2bp relative to the ATG initiation codon drove the transient expression of a luciferase reporter gene when transfected into HRE H9 rabbit endometrial cells. PMA treatment of HRE H9 cells enhanced luciferase transcription approximately threefold. These data suggest that the TRAP promoter is complex and contains multiple regulatory elements. The availability of the TRAP promoter may also permit production of transgenic mice, which can be used to develop previously unavailable osteoclast cell lines.

Quantitative studies on aluminum deposition and its effects on neurofilament protein expression and phosphorylation, following the intraventricular administration of aluminum maltolate to adult rabbits.

The deposition of aluminum (Al) in the brain and spinal cord of adult male New Zealand white rabbits was monitored following the intraventricular administration of Al maltolate. Although decreasing concentrations of Al were observed from the injection site (approximately 10 micrograms/g dry tissue) to the lumbar cord (2.1 micrograms/g), argyrophilic tangles were present in the perikarya and proximal neurites of neurons as far distal as the lumbar and sacral cord areas. Quantitative immunoblot studies of the three neurofilament protein isoforms failed to detect changes resulting from Al maltolate treatment. Similarly no significant alterations in the total phosphate content of these cytoskeletal proteins were observed. Lastly, on Northern blots, the expression of genes encoding for the 200 kDa and 68 kDa neurofilament proteins also was unaffected by Al maltolate treatment.

The effectiveness of weight-belts during multiple repetitions of the squat exercise.

The purpose of this study was to examine the effectiveness of weight-belts during multiple repetitions of the parallel back squat exercise. Five subjects were filmed (50 fps) as they performed eight consecutive trials at each of two weight-belt conditions [with belt = WB, without belt = WOB] in random order at their eight-repetition maximum effort. Other parameters examined were ground reaction forces, intra-abdominal pressure (IAP), and mean electromyography (mEMG) for the external oblique (EO), erector spinae (ES), vastus lateralis (VL), and bicep femoris (BF) muscles. All parameters were collected and interfaced to a computer via an A/D converter. WB repetitions were generally performed faster than WOB repetitions, especially by the later repetitions (3.34 vs 3.56 s). WB IAP values were consistently greater (P less than 0.05) than WOB values by 25-40%. IAP increased by approximately 11.5% from the first to the last repetitions. No differences were observed for ES and EO mEMG for belt usage, but values increased by up to 20% across repetitions. Several differences were observed between WB and WOB for the VL and BF mEMG, with WB values being significantly greater. These data suggest that a weight-belt aids in supporting the trunk by increasing IAP, and that any differential effect due to wearing a weight-belt did not occur over eight repetitions.

Pet project. The use of pet facilitated therapy among the chronically mentally ill.

1. The effects of animal companions on people can be of therapeutic benefit. Using pet facilitated therapy with the chronically mentally ill as a treatment modality offers great potential for positive patient care outcomes. 2. Initiating a pet facilitated therapy program involves establishing policy and procedures regarding animals at the facility. The personality and animal experiences of the patient must be taken into consideration for pet selection. 3. Evaluate the effectiveness of the pet facilitated therapy program. Assess your patient care outcomes to design further interventions.