A common mechanism of chromosomal translocation in T- and B-cell neoplasia.

The chromosomal breakpoint involved in the t(8;14)(q24;q11) chromosome translocation in the SKW-3 cell line, which directly involves the 3' flanking region of the c-myc gene, was cloned and sequenced. The breakpoint on chromosome 8 mapped to a position 3 kb 3' of c-myc while the chromosome 14 breakpoint occurred 36 kb 5' of the gene for the constant region of the alpha chain of the T-cell receptor (TCR). The translocation resulted in a precise rearrangement of sequences on chromosome 8 and what appears to be a functional J alpha segment on chromosome 14. Signal sequences for V-J joining occurred at the breakpoint positions on both chromosomes 14 and 8, suggesting that the translocation occurs during TCR gene rearrangement and that it is catalyzed by the enzymatic systems involved in V-J joining reactions. The involvement of c-myc in the translocation and the association of joining signals at the breakpoints provides a parallel to the situation observed in the translocations involving c-myc and the immunoglobulin loci in B-cell neoplasms and suggests that common mechanisms of translocation and oncogene deregulation are involved in B- and T-cell malignancies.

Advancing the spontaneous hypertensive rat model of attention deficit/hyperactivity disorder.

To advance the spontaneous hypertensive rat (SHR) model of attention deficit/hyperactivity disorder (ADHD), experiments examined the SHR in tasks recognized to assess functioning of the prefrontal cortex or dorsal striatal. Tasks included odor-delayed win-shift (nonspatial working and reference memory), win-stay (habit learning), and attentional set-shifting (attention and behavioral flexibility). In Experiment 1, the SHR strain was compared with Wistar-Kyoto (WKY) and Wistar-Kyoto Hypertensive (WKHT) strains on the first 2 tasks. In Experiment 2, oral methylphenidate (1.5 mg/kg) and vehicle (water) were evaluated on all 3 tasks in SHR and WKY strains. Results demonstrated that the SHR made significantly more errors in the odor-delayed win-shift, win-stay, and attentional set-shifting tasks compared with the WKY. Similar performances in the WKY and WKHT indicated that deficits observed in the SHR were not related solely to hypertension. Treating the SHR with methylphenidate eliminated strain differences in all 3 tasks. These findings provide evidence that the SHR is a valid model for studying ADHD-associated neurocognitive deficits. Moreover, the current behavioral approach is appropriate to assess novel medications developed to target ADHD-associated neurocognitive deficits.

Characterization of a candidate bcl-1 gene.

The t(11;14)(q13;q32) translocation has been associated with human B-lymphocytic malignancy. Several examples of this translocation have been cloned, documenting that this abnormality joins the immunoglobulin heavy-chain gene to the bcl-1 locus on chromosome 11. However, the identification of the bcl-1 gene, a putative dominant oncogene, has been elusive. In this work, we have isolated genomic clones covering 120 kb of the bcl-1 locus. Probes from the region of an HpaII-tiny-fragment island identified a candidate bcl-1 gene. cDNAs representing the bcl-1 mRNA were cloned from three cell lines, two with the translocation. The deduced amino acid sequence from these clones showed bcl-1 to be a member of the cyclin gene family. In addition, our analysis of expression of bcl-1 in an extensive panel of human cell lines showed it to be widely expressed except in lymphoid or myeloid lineages. This observation may provide a molecular basis for distinct modes of cell cycle control in different mammalian tissues. Activation of the bcl-1 gene may be oncogenic by directly altering progression through the cell cycle.

Translocations and rearrangements in T-cell acute leukemias with the t(11;14) (p13;q11) chromosomal translocations.

Chromosomal translocations involving the T-cell receptor alpha and delta genes at q11 on chromosome 14 are the most common cytogenetic abnormalities in patients with T-cell tumors. We have demonstrated that the t(11;14)(p13;q11) translocation in two T-ALL patients involves the J delta region suggesting that the translocation proceeds or coincides with delta gene rearrangement. Additional rearrangements on both normal and translocated chromosomes 14 are described including rearrangement of both Tcr-alpha and Tcr-delta genes, and deletions within the J alpha region. The polyclonality of rearrangements on the normal chromosome 14 in one of the patient samples demonstrates that T-cell receptor rearrangement continues after the translocation event. The identification of clonally expanded rearrangements involving both the Tcr-delta and the Tcr-alpha genes in a single patient suggests a cascade model for delta----alpha expression may be a viable pathway for T-cell maturation.

Transcription-mediated amplification and hybridisation protection assay to determine BCR-ABL transcript levels in patients with chronic myeloid leukaemia.

Detection of BCR-ABL transcripts in chronic myeloid leukaemia (CML) is used to confirm the diagnosis and to monitor residual disease. Quantitative techniques are required to predict response to therapy or early relapse. We have evaluated an assay in which transcription-mediated amplification (TMA) of BCR-ABL and ABL transcripts is achieved using reverse transcriptase and RNA polymerase. The products are quantified in the hybridisation protection assay (HPA) using acridinium ester-labelled DNA probes and chemiluminescence. The method is a single tube procedure which uses small amounts of RNA (<500 ng/triplicate analysis), is technically simple (requiring just two waterbaths and a luminometer), rapid (total assay time <4 h) and sensitive (capable of detecting one BCR-ABL-positive K562 cell in the presence of 10(4)-10(5) BCR-ABL-negative cells). BCR-ABL signals from patient RNA samples were quantified relative to known amounts of K562 RNA and normalised to levels of ABL. BCR-ABL/ABL ratios ranged from 0.15 to 1.59 (median 0.65) in RNA from diagnostic blood or bone marrow of 18 CML patients and were < or =0.0001 in 20 normal controls. Sequential samples analysed from six CML patients post-allogeneic bone marrow transplantation who relapsed and received donor lymphocyte infusions showed BCR-ABL/ABL ratios which reflected patient status or treatment. A BCR-ABL/ABL ratio of 0.01 served as a useful arbitrary indicator value, with results above and below this value generally correlating with relapse or remission, respectively.

Effects of chemical and physical mutagens on the frequency of a large genetic duplication in Salmonella typhimurium. I. Induction of duplications.

In Salmonella typhimurium a simple selection has been described to detect bacteria that are merodiploid for almost one-third of the chromosome. The selective procedure is based upon improved utilization of L-malate as the sole carbon source in merodiploid strains. The spontaneous frequency of the duplication in haploid strains is approximately 10(-4) per cell plated. Following the exposure of a haploid strain to mutagenic agents, there is a dose-dependent increase in the duplication frequency above the spontaneous level. In this paper we describe the induction of genetic duplications in Salmonella typhimurium by X-rays, ultraviolet light (UV), ethyl methanesulfonate (EMS), nitrous acid, and the azaacridine half mustard, ICR-372.

Mutagenesis at the ad-3A and ad-3B loci in haploid UV-sensitive strains of Neurospora crassa. V. Comparison of dose--response curves of single- and double-mutant strains with wild-type.

The interactions of mutant alleles that individually confer radiation sensitivity in Neurospora crassa are being studied with regard to their effects on radiation-induced inactivation and forward-mutation induction at the ad-3 loci. This paper reports attempts to construct 3 double-mutant strains containing the following pair-wise combinations of repair-deficient mutants: upr-1,uvs-2; uvs-2,uvs-6; and uvs-3,uvs-6. The double-mutant strain with the 2 excision-repair-deficient mutants upr-1 and uvs-2 shows increased sensitivity to X-ray-induced mutagenesis and inactivation, relative to that shown by either of the parental single-mutant strains. This double mutant is no more sensitive than the parental single-mutant strains to either UV mutagenesis or inactivation. The combination of the uvs-2 and uvs-6 double-mutant strain is considerably more sensitive to both UV and X-ray inactivation than either the uvs-2 or uvs-6 strain, but it shows no greater sensitivity than the parental strains to ad-3 mutation induction by either agent. The combination of the uvs-3 and uvs-6 alleles is inviable. Tetrad analysis and microscopical examination of ascospores shows that ascospores of presumptive genotype uvs-3, uvs-6 do not grow beyond the formation of a few hyphal threads. The lethal and mutagenic effects of UV and X-irradiation in these double-mutant strains are interpreted in terms of the repair systems in Neurospora and other microorganisms.

The effect of air abrasive polishing on blood pH and electrolyte concentrations in healthy mongrel dogs.

The recent re-emergence of air abrasive polishing as an adjunct to professional tooth cleaning has been accompanied by a substantial amount of research. Most of this research has focused on the efficacy of the Prophy-Jet and similar devices in the removal of plaque and stain and on the effects of such devices on restorative materials and the various hard structures of the tooth. Little attention has been paid to the effects of mucosal absorption of the sodium bicarbonate in the polishing abrasive. The purpose of this study was to measure any changes in arterial blood chemistry resulting from the use of the Prophy-Jet. The values for pH, sodium, and bicarbonate, as well as other electrolytes, were measured in 10 dogs over a 2 hour period following a 5 minute timed use of the Prophy-Jet. Of all the parameters examined, only the potassium concentration showed a statistically significant change, and the magnitude of this change was not felt to be clinically significant. The pH and concentrations of sodium and bicarbonate remained essentially unchanged with the use of the Prophy-Jet.

Cardiopulmonary effects of sevoflurane in Garnett's greater bush baby (Otolemur garnettii).

PURPOSE: A study was conducted to assess the cardiopulmonary and anesthetic effects of sevoflurane anesthesia on Garnett's Greater Bush Baby (Otolemur garnettii). METHODS: Anesthesia was induced in ten animals with 8% sevoflurane and was maintained by use of 2.5% sevoflurane for 30 minutes. Induction and recovery times were recorded. Heart and respiratory rates (RR), end-tidal carbon dioxide concentration (ET CO2), arterial blood pressures, relative arterial blood oxygen saturation (SpO2), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), and pH were monitored. Pre- and poststudy CBC and serum biochemical values were compared. RESULTS: Anesthesia induction was rapid (75+/-8.7 seconds [mean +/- SEM]) and smooth. Heart rate significantly increased initially, then decreased significantly over the remaining 30 minutes. There were no significant changes in RR, SpO2, ETCO2, or arterial blood pressure. The PaO2 values significantly increased in the 10- to 30-minute samples. The PaCO2 values remained steady in the 10- to 30-minute samples. A significant decrease was seen in white blood count, calcium, and total protein (TP) values, compared with values in pre-anesthesia samples. Recovery from anesthesia was smooth and rapid, with extubation at 24+/-5.8 seconds. CONCLUSIONS: At the concentrations used in this study, sevoflurane appears to be a safe and effective agent for induction and maintenance of anesthesia in O. garnettii.

Cardiovascular effects of the alpha2-adrenergic receptor agonist medetomidine in clinically normal cats anesthetized with isoflurane.

OBJECTIVE: To assess the cardiovascular effects of the alpha2-adrenergic receptor agonist medetomidine in healthy cats anesthetized with 2% isoflurane. ANIMALS: 11 clinically normal cats. PROCEDURE: Cats were anesthetized with isoflurane, and catheters were inserted for measurement of aortic, left ventricular, and right atrial pressures. For data collection, end-tidal isoflurane concentration was reduced to 2%, and end-tidal CO2 was maintained at 35 to 40 mm of Hg by use of positive-pressure ventilation. After measurement of baseline data, medetomidine (0.01 mg/kg of body weight, i.m.) was administered and data were collected continuously for 75 minutes. At the end of data collection, incisions were closed and cats were allowed to recover from anesthesia. RESULTS: Medetomidine significantly increased mean arterial pressure and systemic vascular resistance. The increase in mean arterial pressure was maximal at 17.8 +/- 7 minutes after medetomidine administration. Medetomidine also increased left ventricular peak systolic pressure, left ventricular end diastolic pressure, and right atrial pressure. Medetomidine significantly decreased heart rate and mean aortic flow. CONCLUSIONS: The low dosage of medetomidine (0.01 mg/kg, i.m.) promoted severe vasoconstriction in isoflurane-anesthetized cats, and resulted in sustained increases in left ventricular preload and afterload.

Otoacoustic emission in a dog.

A 7-month-old female Poodle was evaluated for a sound continuously emanating from its left ear. The frequency and intensity of the tone was matched by use of a multifrequency audiometer. The frequency and intensity of the sound were 9,594 Hz and 45.5 decibels sound pressure level, respectively. On the bases of normal brain stem auditory-evoked responses and behavioral response to sound, the dog was believed to have normal hearing in both ears. Emission of sound from an ear, referred to as an otoacoustic emission, is a condition that has been reported in human beings and small animals. Successful treatment in human patients with drugs and masking sounds is difficult, and treatment has not been reported in animals. This condition in dogs is not considered life-threatening.

Precautions when using mask induction.

There are few techniques as frequently employed in veterinary anesthesia that are more dependent on good clinical judgment, technical skill, and finesse than mask induction. The technique can be extremely valuable in selected patients.

Critical care surgical techniques.

A veterinarian dealing with critical and trauma patients must be proficient with techniques for tracheostomy, thoracostomy tube placement for chest drainage, diagnostic peritoneal lavage, and autotransfusion. The utilization of these techniques may be life-saving in the critical patient. A tracheostomy is indicated in any patient with upper airway obstruction that cannot be managed with supplemental oxygen and/or orotracheal intubation. A tracheostomy tube with an inner cannula is preferred. Tracheostomy tubes should be cleaned at 3- to 4-h intervals, and methods should be employed to decrease thick tracheal secretions and to remove them from the trachea. A patient with a tracheostomy tube should be monitored continuously. A thoracostomy tube is indicated in any patient with large and/or continuous accumulation of air, blood, fluid, or chyle in the pleural space. The thoracostomy tube should be at least the same size as the patient's main stem bronchus. The thoracostomy tube is placed aseptically at the seventh intercostal space at the junction of the upper one third and lower two thirds of the lateral chest wall. Fluid or air may be removed from the chest intermittently with a three-way stopcock attached to the thoracostomy tube and a 60-ml syringe. If continuous drainage is needed, a continuous underwater seal and suction system should be used. Diagnostic abdominal paracentesis and peritoneal lavage are useful techniques in the determination of abdominal trauma, hollow viscus rupture, peritonitis, hepatic trauma, and urinary system trauma. When a multiholed catheter and lavage are used, the accuracy of detecting abdominal trauma is 95 per cent. When only needle paracentesis is used, the accuracy drops to 47 per cent. Abdominal lavage fluid can be analyzed for bacteria, whole blood, white blood cells, free bilirubin, creatinine, blood urea nitrogen, amylase, alkaline phosphatase, and alanine aminotransferase. Large volumes of whole blood recovered from abdominal or thoracic paracentesis can be reinfused into the patient if needed, providing it is not contaminated or markedly hemolyzed. The blood should be collected aseptically into blood bottles or bags. If the bleeding is ongoing or the blood only a few hours old, anticoagulants should be used. If the hemorrhage is several hours old, then clotting and defibrination has already occurred and the blood can be collected into "dry" bags or bottles. Before use, abdominal blood should be analyzed for urine, bile or fecal contamination. Blood collected from the thoracic cavity is much less likely to be contaminated. Autotransfused blood is administered through a standard blood administration set.

Management of geriatric patients. A common occurrence.

The anesthetic and perioperative care of geriatric animals requires increased vigilance and support. The margin of acceptable physiological variation is probably more narrow than in younger patients. Underlying disease, which is often subclinical, influences metabolism and recovery from anesthetics and also predisposes the patients to adverse outcome. Limited respiratory and cardiovascular reserve diminishes the ability of many older patients to meet the challenges of anesthesia and surgery or other stressful medical procedures. The psychological, as well as physiological, stress of hospitalization is increased in many geriatric patients.

Alpha 2 agonists and antagonists.

The alpha 2 agonists can produce reliable dose-dependent sedation and analgesia in most species. Nevertheless, they can also produce significant physiological adverse side effects depending on dose, rate, route of administration, and the concurrent use of other CNS depressants. For this reason, it may be best to use a low dose of an alpha 2 agonist as a preanesthetic agent. The alpha 2 agonists are best suited for young, healthy, exercise-tolerant patients. The combining of low doses of alpha 2, opioid, and benzodiazepine agonists results in a synergistic CNS depressant response while minimizing the undesirable side effects of these three classes of drugs. Each group of drugs has specific antagonists available for their reversal, thus allowing veterinarians to reverse one or more of the agonists depending on the desired response. This may represent a significant advantage to the use of low-dose alpha 2 agonists in combination with opioids and benzodiazepines.

Cardiopulmonary and anesthetic effects of medetomidine-ketamine-butorphanol and antagonism with atipamezole in servals (Felis serval).

Seven (three male and four female) 4-7-yr old captive servals (Felis serval) weighing 13.7 +/- 2.3 kg were used to evaluate the cardiopulmonary and anesthetic effects of combined intramuscular injections of medetomidine (47.4 +/- 10.3 microg/kg), ketamine (1.0 +/- 0.2 mg/kg), and butorphanol (0.2 +/- 0.03 mg/kg). Inductions were smooth and rapid (11.7 +/- 4.3 min) and resulted in good muscle relaxation. Significant decreases in heart rate (85 +/- 12 beats/min) at 10 min after injection and respiratory rate (27 +/- 10 breaths/min) at 5 min after injection continued throughout the immobilization period. Rectal temperature and arterial blood pressure did not change significantly. The PaO2 decreased significantly, and PaCO2 increased significantly during immobilization but remained within clinically acceptable limits. Hypoxemia (PaO2 < 60 mm Hg) was not noted, and arterial blood oxygen saturation (SaO2) was greater than 90% at all times. Relative arterial oxygen saturation (SpO2) values, indicated by pulse oximetry, were lower than SaO2 values. All animals could be safely handled while sedated. Administration of atipamezole (236.8 +/- 51.2 microg/kg half i.v. and half s.c.), an alpha2 antagonist, resulted in rapid (4.1 +/- 3 min to standing) and smooth recoveries.

Predictors of early mortality among hospitalized nursing home residents.

BACKGROUND: Emergency admissions from nursing homes (NHs) are associated with high mortality. Understanding the predictors of early mortality in these patients may guide clinicians in choosing appropriate site and level of care. METHODS: We identified all consecutive admissions from NHs (all ages) to an Acute Medical Assessment Unit between January 2005 and December 2007. Analysis was performed at the level of the admission. The predictors of in-patient mortality at 7 days were examined using a generalized estimating equations analysis. RESULTS: A total of 314 patients [32% male, mean age: 84.2 years (SD: 8.3 years)] were admitted during the study period constituting 410 emergency episodes. Twenty-three percent of admissions resulted in hospital mortality with 73% of deaths occurring within 1 week (50% within the first 3 days). For 7-day mortality outcome, patients with a modified early warning score (MEWS) of 4-5 on admission had 12 times the odds of death [95% confidence interval (CI) 1.40-103.56], whereas those with a score of ≥6 had 21 times the odds of death (95% CI 2.71-170.57) compared with those with a score of ≤1. An estimated glomerular filtration rate (eGFR) of 30-60 and <30 ml/min/m(2) was associated with nearly a 3-fold increase in the odds of death at 1 week (95% CI 1.10-7.97) and a 5-fold increase in the odds of death within 1 week (95% CI 1.75-14.96), respectively, compared with eGFR > 60 ml/min/m(2). C-reactive protein (CRP) >100 mg/l on admission was also associated with a 2.5 times higher odds of death (95% CI 1.23-4.95). Taking eight or more different medication items per day was associated with only a third of the odds of death (95% CI 0.09-0.98) compared with patients taking only three or fewer per day. CONCLUSION: In acutely ill NH residents, MEWS is an important predictor of early hospital mortality and can be used in both the community and the hospital settings to identify patients whose death maybe predictable or unavoidable, thus allowing a more holistic approach to management with discussion with patient and relatives for planning of immediate care. In addition, CRP and eGFR levels on admission have also been shown to predict early hospital mortality in these patients and can be used in conjunction with MEWS in the same way to allow decision making on the appropriate level of care at the point of hospital admission.

GSI-I (Z-LLNle-CHO) inhibits γ-secretase and the proteosome to trigger cell death in precursor-B acute lymphoblastic leukemia.

Gamma secretase inhibitors (GSIs) comprise a growing class of compounds that interfere with the membrane-bound Notch signaling protein and its downstream intra-nuclear transcriptional targets. As GSI-I (Z-LLNle-CHO) is also a derivative of a widely used proteosome inhibitor MG-132, we hypothesized that this compound might be active in precursor-B acute lymphoblastic leukemia (ALL) cell lines and patient samples. We found that GSI-I treatment of precursor-B ALL blasts induced apoptotic cell death within 18-24 h. With confirmation using RNA and protein analyses, GSI-I blocked nuclear accumulation of cleaved Notch1 and Notch2, and inhibited Notch targets Hey2 and Myc. Microarray analyses of 207 children with high-risk precursor-B ALL demonstrate that Notch pathway expression is a common feature of these neoplasms. However, microarray studies also implicated additional transcriptional targets in GSI-I-dependent cell death, including genes in the unfolded protein response, nuclear factor-κB and p53 pathways. Z-LLNle-CHO blocks both γ-secretase and proteosome activity, inducing more robust cell death in precursor-B ALL cells than either proteosome-selective or γ-secretase-selective inhibitors alone. Using Z-LLNle-CHO in a nonobese diabetes/severe combined immunodeficiency (NOD/SCID) precursor-B ALL xenograft model, we found that GSI-I alone delayed or prevented engraftment of B-lymphoblasts in 50% of the animals comprising the experimental group, suggesting that this compound is worthy of additional testing.