MRI of CNS lymphoma with choroid plexus involvement in five dogs and one cat.

OBJECTIVES: To describe the clinical features, magnetic resonance imaging (MRI) findings, and outcome of dogs and cats with central nervous system (CNS) lymphoma that involved the choroid plexus. MATERIALS AND METHODS: A bi-institutional retrospective study of MRI of dogs and cats with CNS lymphoma, in which the choroid plexus was affected on MRI. Signalment, clinical, MRI, clinicopathologic and histopathologic findings were recorded and evaluated. RESULTS: CNS lymphoma with choroid plexus involvement on the MRI was identified in five dogs and one cat. MRI revealed diffuse enlargement and multifocal nodularity in the choroid plexus in most cases, with the fourth ventricle the most common site affected. Five of the cases had signs of extraneural involvement (including the cat), while the sixth case was not staged. Four of five CSF samples analysed provided a diagnosis of lymphoma. CLINICAL SIGNIFICANCE: We report MRI findings of CNS lymphoma involving the choroid plexus. These results show the importance of recognising novel imaging patterns and the potential utility of CSF collection in diagnosing CNS lymphoma involving the choroid plexus ante mortem.

Force plate gait analysis in Doberman Pinschers with and without cervical spondylomyelopathy.

BACKGROUND: The most accepted means of evaluating the response of a patient with cervical spondylomyelopathy (CSM) to treatment is subjective and based on the owner and clinician's perception of the gait. OBJECTIVE: To establish and compare kinetic parameters based on force plate gait analysis between normal and CSM-affected Dobermans. ANIMALS: Nineteen Doberman Pinschers: 10 clinically normal and 9 with CSM. METHODS: Force plate analysis was prospectively performed in all dogs. At least 4 runs of ipsilateral limbs were collected from each dog. Eight force platform parameters were evaluated, including peak vertical force (PVF) and peak vertical impulse (PVI), peak mediolateral force (PMLF) and peak mediolateral impulse, peak braking force and peak braking impulse, and peak propulsive force (PPF) and peak propulsive impulse. In addition, the coefficient of variation (CV) for each limb was calculated for each parameter. Data analysis was performed by a repeated measures approach. RESULTS: PMLF (P = .0062), PVI (P = .0225), and PPF (P = .0408) were found to be lower in CSM-affected dogs compared with normal dogs. Analysis by CV as the outcome indicated more variability in PVF in CSM-affected dogs (P = 0.0045). The largest difference in the CV of PVF was seen in the thoracic limbs of affected dogs when compared with the thoracic limbs of normal dogs (P = 0.0019). CONCLUSIONS AND CLINICAL IMPORTANCE: The CV of PVF in all 4 limbs, especially the thoracic limbs, distinguished clinically normal Dobermans from those with CSM. Other kinetic parameters less reliably distinguished CSM-affected from clinically normal Dobermans.

Three-dimensional kinematic gait analysis of Doberman Pinschers with and without cervical spondylomyelopathy.

BACKGROUND: The optimal treatment of cervical spondylomyelopathy (CSM) is controversial, with the owner's and clinician's perception of gait improvement often being used as outcome measures. These methods are subjective and suffer from observer bias. OBJECTIVES: To establish kinematic gait parameters utilizing digital motion capture in normal Doberman Pinschers and compare them with CSM-affected Dobermans. ANIMALS: Nineteen Doberman Pinschers; 10 clinically normal and 9 with CSM. METHODS: All dogs were enrolled prospectively and fitted with a Lycra(®) body suit, and motion capture was performed and used to reconstruct a 3-D stick diagram representation of each dog based on 32 reflective markers, from which several parameters were measured. These included stride duration, length, and height; maximal and minimal spinal angles; elbow and stifle flexion and extension; and maximum and minimum distances between the thoracic and pelvic limbs. A random-effects linear regression model was used to compare parameters between groups. RESULTS: Significant differences between groups included smaller minimum (mean = 116 mm; P = .024) and maximum (mean = 184 mm; P = .001) distance between the thoracic limbs in CSM-affected dogs. Additionally, thoracic limb stride duration was also smaller (P = .009) in CSM-affected dogs (mean = 0.7 seconds) when compared with normal dogs (mean = 0.8 seconds). In the pelvic limbs, the average stifle flexion (mean = 100°; P = .048) and extension (mean = 136°; P = .009), as well as number of strides (mean = 2.7 strides; P = .033) were different between groups. CONCLUSIONS AND CLINICAL IMPORTANCE: Our findings suggest that computerized gait analysis reveals more consistent kinematic differences in the thoracic limbs, which can be used as future objective outcome measures.

Sex-related injury patterns among selected high school sports.

This cohort observational study was undertaken to test the hypothesis that the incidence of injuries for girls participating in high school sports is greater than that for boys. From 1995 through 1997, players were included in our study if they were listed on the school's varsity team roster for boys' or girls' basketball, boys' or girls' soccer, boys' baseball, or girls' softball. Injuries and opportunities for injury were recorded daily. Certified athletic trainers reported injury and exposure data. Based on 39,032 player-seasons and 8988 reported injuries, the injury rates per 100 players for softball (16.7) and for girls' soccer (26.7) were higher than for baseball (13.2) and boys' soccer (23.4). The knee injury rates per 100 players for girls' basketball (4.5) and girls' soccer (5.2) were higher than for their male counterparts. Major injuries occurred more often in girls' basketball (12.4%) and soccer (12.1%) than in boys' basketball (9.9%) and soccer (10.4%). Baseball players (12.5%) had more major injuries than softball players (7.8%). There was a higher number of surgeries, particularly knee and anterior cruciate ligament surgeries, for female basketball and soccer players than for boys or girls in other sports.

Traumatic brain injury in high school athletes.

CONTEXT: The potential seriousness of mild traumatic brain injury (MTBI) is increasingly recognized; however, information on the frequency of MTBI among high school athletes is limited. OBJECTIVE: To identify the type, frequency, and severity of MTBI in selected high school sports activities. DESIGN: Observational cohort study. SETTING AND PARTICIPANTS: Two hundred forty-six certified athletic trainers recorded injury and exposure data for high school varsity athletes participating in boys' football, wrestling, baseball and field hockey, girls' volleyball and softball, boys' and girls' basketball, and boys' and girls' soccer at 235 US high schools during 1 or more of the 1995-1997 academic years. MAIN OUTCOME MEASURES: Rates of reported MTBI, defined as a head-injured player who was removed from participation and evaluated by an athletic trainer or physician prior to returning to participation. National incidence figures for MTBI also were estimated. RESULTS: Of 23566 reported injuries in the 10 sports during the 3-year study period, 1219 (5.5%) were MTBIs. Of the MTBIs, football accounted for 773 (63.4%) of cases; wrestling, 128 (10.5%); girls' soccer, 76 (6.2%); boys' soccer, 69 (5.7%); girls' basketball, 63 (5.2%); boys' basketball, 51 (4.2%); softball, 25 (2.1%); baseball, 15 (1.2%); field hockey, 13 (1.1%); and volleyball, 6 (0.5%). The injury rates per 100 player-seasons were 3.66 for football, 1.58 for wrestling, 1.14 for girls' soccer, 1.04 for girls' basketball, 0.92 for boys' soccer, 0.75 for boys' basketball, 0.46 for softball, 0.46 for field hockey, 0.23 for baseball, and 0.14 for volleyball. The median time lost from participation for all MTBIs was 3 days. There were 6 cases of subdural hematoma and intracranial injury reported in football. Based on these data, an estimated 62816 cases of MTBI occur annually among high school varsity athletes participating in these sports, with football accounting for about 63% of cases. CONCLUSIONS: Rates of MTBI vary among sports and none of the 10 popular high school sports we studied is without the occurrence of an MTBI. Continued involvement of high school sports sponsors, researchers, medical professionals, coaches, and sports participants is essential to help minimize the risk of MTBI.

Injury patterns in selected high school sports: a review of the 1995-1997 seasons.

OBJECTIVE: To characterize the risk of injury associated with 10 popular high school sports by comparing the relative frequency of injury and selected injury rates among sports, as well as the participation conditions within each sport. DESIGN AND SETTING: A cohort observational study of high school athletes using a surveillance protocol whereby certified athletic trainers recorded data during the 1995-1997 academic years. SUBJECTS: Players listed on the school's varsity team rosters for football, wrestling, baseball, field hockey, softball, girls' volleyball, boys' or girls' basketball, and boys' or girls' soccer. MEASUREMENTS: Injuries and opportunities for injury (exposures) were recorded daily. The definition of reportable injury used in the study required that certified athletic trainers evaluate the injured players and subsequently restrict them from participation. RESULTS: Football had the highest injury rate per 1000 athlete- exposures at 8.1, and volleyball had the lowest rate at 1.7. Only boys' (59.3%) and girls' (57.0%) soccer showed a larger proportion of reported injuries for games than practices, while volleyball was the only sport to demonstrate a higher injury rate per 1000 athlete-exposures for practices than for games. More than 73% of the injuries restricted players for fewer than 8 days. The proportion of knee injuries was highest for girls' soccer (19.4%) and lowest for baseball (10.5%). Among the studied sports, sprains and strains accounted for more than 50% of the injuries, except in field hockey (45.7%). Of the injuries requiring surgery, 60.3% were to the knee. CONCLUSIONS: An inherent risk of injury is associated with participation in high school sports based on the nature of the game and the activities of the players. Therefore, injury prevention programs should be in place for both practices and games. Preventing reinjury through daily injury management is a critical component of an injury prevention program. Although sports injuries cannot be entirely eliminated, consistent and professional evaluation of yearly injury patterns can provide focus for the development and evaluation of injury prevention strategies.

A Canadian hospital-based HIV/hepatitis C look-back notification program.

OBJECTIVE: To describe the process used to notify pediatric patients who received transfusions of blood or blood products at our institution before donor blood was routinely screened for antibodies to HIV (1985) and hepatitis C virus (1990), and to evaluate the effectiveness of the notification program. DESIGN: Patients who had received transfusions were identified through the hospital's medical records and the records from the Transfusion Medicine Laboratory. Patients were contacted by registered mail to provide notification of transfusion. A questionnaire was included with the notification to obtain information about the patient's awareness of the transfusion and whether he or she had undergone or planned to undergo testing for HIV and hepatitis C virus. SETTING: Tertiary care university-affiliated teaching hospital in Hamilton, Ont. PATIENTS: Patients 16 years of age or younger who had received blood products between February 1978 and November 1985. Patients who had received only albumin or immune serum globulin were not included as these products were not associated with viral transmission in Canada. RESULTS: Notification letters were sent to 1546 patients. Of these letters 522 (33.8%) were returned undelivered. Of the 1024 patients contacted 493 (48.1%) responded to the questionnaire, of whom 157 (31.8%) were not aware of their transfusion. A total of 130 (26.4%) of the respondents had already undergone testing for HIV, and 342 (69.4%) indicated that they would undergo such testing as a result of the notification. In contrast, only 30 (6.3%) of 474 respondents had undergone testing for hepatitis C virus, but 425 (89.7%) indicated that they would undergo such testing. Overall, the patients' response to the notification was neutral or positive; however, a number of patients expressed dissatisfaction and anxiety. CONCLUSIONS: The high proportion of patients who were unaware that they had undergone transfusion and who decided to undergo testing for HIV and hepatitis C virus as a result of notification supports the use of notification programs such as this one.

Molecular cloning, upstream sequence and promoter studies of the human gene for the regulatory subunit RII alpha of cAMP-dependent protein kinase.

The gene for the regulatory subunit RII alpha of cAMP-dependent protein kinase is highly regulated during spermatogenesis and a strong signal from a distinct short mRNA form is observed postmeiotically during spermatid elongation. This report presents the isolation and characterization of the 5'-flanking region (1.2 kb) and exon 1 of the human RII alpha gene. S1 nuclease mapping and primer extension experiments revealed the presence of a major transcriptional start site located 208 nucleotides upstream of start for translation. The 5'-flanking region of the RII alpha gene did not contain a TATA box and was highly G/C-rich. A basal promoter directing high levels of chloramphenicol acetyl transferase (CAT) activity was identified in the 5'-flanking sequence. Several potential binding sites for transcription factors were identified in this region, which may be responsible for the germ cell-specific regulation of this gene. We have previously reported that the human testis RII alpha cDNA contains a region (amino acids 45-75) with little or no homology to the corresponding rat skeletal muscle cDNA (Oyen, O., Myklebust, F., Scott, J.D., Cadd, G.G., McKnight, G.S., Hansson, V. and Jahnsen, T. (1990) Biol. Reprod. 43, 46-54). We examined whether this difference could arise due to organ-specific splice mechanisms or represented a species difference. We show that the low homology region of the human RII alpha cDNA resides entirely within exon 1, and does not originate from a tissue-specific alternate splicing of this distinct region.

Acetylsalicylic acid and sodium salicylate inhibit LPS-induced NF-kappa B/c-Rel nuclear translocation, and synthesis of tissue factor (TF) and tumor necrosis factor alfa (TNF-alpha) in human monocytes.

We have investigated the effects of acetylsalicylic acid and sodium salicylate on the LPS-induced synthesis of the pro-coagulant protein tissue factor (TF) and the pro-inflammatory protein tumor necrosis factor-alpha (TNF-alpha), as well as the prostaglandin PGE2 in human monocytes. Both drugs dose-dependently inhibited LPS-induced TF and TNF-alpha synthesis at the mRNA and the protein level, and reduced PGE2 production. As evidenced by electro mobility shift assay (EMSA) and the use of a NF-kappa B prototypic probe, these drugs probably exert their inhibitory effects by interference with the nuclear translocation of NF-kappa B/c-Rel proteins. These data may expand the understanding of the anti-thrombotic and anti-inflammatory effects of these drugs when activation of monocytes occurs.

Structure and function of the mouse surfactant protein B gene.

The mouse surfactant protein B (SP-B) gene was isolated from a DBA/2J EMBL3 genomic library, and the nucleotide sequence and transcriptional start site was determined. The SP-B gene was comprised of 11 exons and 10 introns and spanned 9,882 bases. Repetitive elements of the B1, B2, and R families and several other motifs were located within nontranslated regions of the gene. The 5' flanking region of the gene was utilized to express a bacterial chloramphenicol acetyl transferase reporter gene in mouse and human pulmonary adenocarcinoma cell lines. Transcriptionally active regions of the promoter were identified that determined lung epithelial cell-specific gene expression. Regions of the gene promoter shared between the mouse and human SP-B gene were used to locate putative transcription factor binding sites for thyroid transcription factor-1 and hepatocyte nuclear factor-3. The 5' flanking region of the murine SP-B contains cell-specific cis-active elements that regulate SP-B expression in the respiratory epithelium.

Parathyroid hormone-related protein: a potential autocrine growth regulator in human prostate cancer cell lines.

OBJECTIVE: We recently demonstrated that parathyroid hormone-related protein (PTHrP) is widely expressed by human prostate cancer tissue, suggesting that PTHrP might be involved in the growth and development of prostate cancer. To study this further, the production of PTHrP and its biologic effect were investigated using human prostate cancer cell lines. METHODS: The cell lines used were one androgen-dependent cell line, LNCaP, and two androgen-independent cell lines, PC-3 and DU-145. PTHrP secreted by cancer cells was measured by radioimmunoassay. The effect of PTHrP on DNA synthesis in these cells was determined by thymidine incorporation assay. RESULTS: All cell lines secreted immunodetectable levels of PTHrP in the culture-conditioned media. PC-3 cells secreted significantly higher amounts than the other two cell lines. A synthetic peptide, PTHrP(1-34), stimulated thymidine uptake in PC-3 and DU-145 cells more than threefold the control under serum-free and steroid-free conditions, whereas LNCaP was not affected. However, in the presence of dihydrotestosterone, DNA synthesis of LNCaP cells was stimulated by PTHrP in a dose-dependent manner. Additionally, this PTHrP-induced DNA synthesis was completely neutralized by a validated mouse monoclonal antibody (8B12) raised against PTHrP(1-34). CONCLUSIONS: Our data suggest that PTHrP may play a significant role in the growth of prostate cancer by acting locally in an autocrine fashion.

Characterization of in-vitro-translated human regulatory and catalytic subunits of cAMP-dependent protein kinases.

Full-length human cDNAs for all the different regulatory (R) and catalytic (C) subunits of cAMP-dependent protein kinases (PKA) were transcribed and translated in a cell-free in vitro system. The resulting proteins were characterized with respect to molecular size, isoelectric focusing, immunoreactivity, cAMP binding, and to what extent the RII protein subunits revealed mobility shifts upon phosphorylation by catalytic subunit of PKA. We were able to express cDNAs for all the human R (RI alpha, RI beta, RII alpha and RII beta) and C (C alpha, C beta and C gamma) subunits in a wheat-germ extract. [35S]Methionine-labelled in-vitro-translated products were analyzed by SDS/PAGE and revealed distinct protein bands with apparent molecular masses of 49 (RI alpha), 54-55 (RI beta), 51 (RII alpha) and 53 kDa (RII beta) for the R subunits. In vitro transcription/translation of the cDNAs for the C subunits of PKA gave proteins with molecular masses of approximately 40 kDa for all the different C subunits. Phosphorylation of RII alpha and RII beta by the C subunit of PKA, revealed a distinct mobility shift of the RII alpha subunit on one-dimensional SDS/PAGE (51-54 kDa), but not of RII beta (53 kDa). Further characterization of the R subunits by two-dimensional SDS/PAGE revealed that RI alpha was more acidic than RI beta, with pIs of 6.1-6.0 and 6.4-6.2, respectively. Furthermore, the RII alpha protein was more basic than RII beta, with pIs of approximately 5.4-5.3 and 5.3-5.1, respectively. All the in-vitro-translated R subunits could be photoaffinity labelled by the cAMP-analog 8-azido-[32P]cAMP and were also detected by immunoprecipitation with subunit-specific antibodies.

Expectations and needs of persons with family members in an intensive care unit as opposed to a general ward.

The positive effect of family support on the outcome from serious illness that requires intensive care has been recognized by clinicians for decades. We have all seen that family visitation and an intensive care environment more similar to that of a general ward (sunlight, radio, television) can benefit patients with psychosis related to intensive care. The severity of illness of the individual patient exerts a powerful stress on the family unit, but it has been difficult to measure this effect. We used a 40-question family needs survey with a degree of importance scale to compare the intensive care unit (ICU) with the general ward in terms of impact on the family. Five needs were found to discriminate these two environments. The family members of patients in an ICU considered it very important (1) for staff to give directions on what to do at the bedside, (2) to receive more support from their own family unit, (3) to have a place to be alone as a family unit in the hospital, (4) to be informed in advance of any transfer plan, and (5) to have flexibility in the time allowed for visitation. Family members are willing to accept decreased visitation time if the physicians and nurses can equate this decrease with the complexity of care in the ICU. The results of this survey have helped us modify and individualize our approach based on family expectations especially when patients are transferred from the general ward to the ICU or from the ICU to the ward.

Identification, purification, and characterization of subunits of cAMP-dependent protein kinase in human testis. Reverse mobilities of human RII alpha and RII beta on sodium dodecyl sulfate-polyacrylamide gel electrophoresis compared with rat and bovine RIIs.

We have previously identified and characterized regulatory (R) subunits of cyclic AMP-dependent protein kinase, particularly the RII subunits in rat tissues (Jahnsen, T., Lohmann, S. M., Walter, U., Hedin, L., and Richards, J. S. (1985) J. Biol. Chem. 260, 15980-15987; Jahnsen, T., Hedin, L., Lohmann, S. M., Walter, U., and Richards, J. S. (1986) J. Biol. Chem. 261, 6637-6639; Jahnsen, T., Hedin, L., Kidd, V. J., Beattie, W. G., Lohmann, S. M., Walter, U., Durica, J., Schulz, T. Z., Schiltz, E., Browner, M., Lawrence, C. B., Goldman, D., Ratoosh, S. L., and Richards, J. S. (1986) J. Biol. Chem. 261, 12352-12361). These studies showed that rat RII alpha and RII beta had apparent molecular masses of 54 and 52 kDa, respectively. The aim of the present study was to purify and characterize cAMP-dependent protein kinase R subunits in human testis and to examine which of the subunits (mRNAs and proteins) are present in this tissue. Our results show that human testis contains mRNAs for five out of the seven known subunits of cAMP-dependent protein kinase. We observed strong expression of mRNAs for RI alpha (1.5 and 3.2 kilobases (kb)), RII alpha (2.2, 2.4, and 7.0 kb), and RII beta (3.3 kb). We also demonstrated mRNAs for two of the three catalytic subunits, C alpha (2.7 kb) and C gamma (1.7 kb). Purification of R subunits by DEAE-cellulose and cAMP affinity chromatography revealed three distinct forms with apparent molecular masses of 49, 51, and 53 kDa, respectively. Characterization of these R subunits by their 8-azido-cAMP photoaffinity labeling and immunoreactivity, as well as by a phosphorylation-dependent mobility shift on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), indicated subunit sizes of RII beta (53 kDa) greater than RII alpha dephosphoform (51 kDa) greater than RI alpha (49 kDa). This conclusion was verified by the analysis of RII subunits produced by in vitro transcription/translation of full-length cDNAs for both human RII alpha and RII beta in wheat germ lysates. The in vitro translated products were the same size as the purified human testis subunits, and only the smallest RII subunit (RII alpha) revealed a distinct mobility shift on SDS-PAGE after phosphorylation/dephosphorylation. This study supports the conclusion that the mobilities of human RII subunits (RII alpha, RII beta) on SDS-PAGE are reversed in contrast with those of other species such as rat and bovine.(ABSTRACT TRUNCATED AT 400 WORDS)

Localization of the catalytic subunit C gamma of the cAMP-dependent protein kinase gene (PRKACG) to human chromosome region 9q13.

A cDNA for a new catalytic subunit (C gamma) of the cAMP-dependent protein kinase (PKA) was recently isolated from a human testis cDNA library. This subunit was shown to be expressed only in testis, and has so far not been demonstrated in other species. In the present study, we have determined the chromosomal localization of this gene employing a cDNA for C gamma as a probe. Southern blot analysis of genomic DNA from human x mouse somatic cell hybrids allowed us to assign this gene (PRKACG) to human chromosome 9. In situ hybridization to metaphase chromosomes confirmed the somatic cell hybrid data and regionally mapped the C gamma gene of PKA to human chromosome 9q13.

Four sites in the acceptor helix and one site in the variable pocket of tRNA(Ala) determine the molecule's acceptor identity.

The structural features that determine tRNA(Ala) acceptor identity have been studied with amber-suppressor tRNAs in Escherichia coli cells. Previous work established that a wobble pair composed of guanosine at position 3 and uridine at position 70 (G3-U70) in the acceptor helix of tRNA(Ala) is a determinant of the molecule's acceptor identity. We show that additional determinants are located at three other sites in the acceptor helix and at one site in the variable pocket of tRNA(Ala). These latter determinants are less important than G3.U70 since their individual alterations in mutants of tRNA(Ala) have smaller degrading effects on the functions of the molecules, and subsets of the determinants, when combined with G3.U70, are sufficient to switch the identities of several other tRNAs to that of tRNA(Ala). Other workers are using fragments of the tRNA(Ala) acceptor helix to study the molecule's acceptor identity. Our demonstration that the variable pocket contributes to tRNA(Ala) acceptor identity means that such fragments do not faithfully replicate the structure-function relationship of the cellular process.

Rapid determination of nucleotides that define tRNA(Gly) acceptor identity.

Expression of the genetic code depends on the recognition of specific tRNAs by the enzymes that aminoacylate them. A computer comparison of tRNA sequences coupled with analysis of mutant nonsense-suppressor tRNAs has revealed the structural features that distinguish the acceptor identity of Escherichia coli tRNA(Gly) from tRNAs that accept phenylalanine, arginine, lysine, and glutamine. On replacement of several nucleotides in the acceptor stem and anticodon of the latter tRNAs with tRNA(Gly)-derived residues, the resulting molecules acquired a tRNA(Gly) identity.