Nitric oxide amplifies interleukin 1-induced cyclooxygenase-2 expression in rat mesangial cells.

Interleukin 1 and nitric oxide (NO) from infiltrating macrophages and activated mesangial cells may act in concert to sustain and promote glomerular damage. To evaluate if such synergy occurs, we evaluated the effect if IL-1 beta and NO on the formation of prostaglandin (PG)E2 and cyclooxygenase (COX) expression. The NO donors, sodium nitroprusside and S-nitroso-N-acetylpenicillamine, alone did not increase basal PGE2 formation. However, these compounds amplified IL-1 beta-induced PGE2 production. Similarly, sodium nitroprusside and S-nitroso-N-acetylpenicillamine by themselves did not induce mRNA and protein for COX-2, the inducible isoform of COX; however, they both potentiated IL-1 beta-induced mRNA and protein expression of COX-2. The stimulatory effect of NO is likely to be mediated by cGMP since (a) an inhibitor of the soluble guanylate cyclase, methylene blue, reversed the stimulatory effect of NO donors on COX-2 mRNA expression; (b) the membrane-permeable cGMP analogue, 8-Br-cGMP, mimicked the stimulatory effect of NO donors on COX-2-mRNA expression; and (c) atrial natriuretic peptide, which increases cellular cGMP by activating the membrane-bound guanylate cyclase, also amplified IL-1 beta-induced COX-2 mRNA expression. These data indicate a novel interaction between NO and COX pathways.

Model for the genetic evolution of human solid tumors.

A conceptual model is proposed for the genetic evolution of many human solid tumors that is based on the observations that cancer cells may spontaneously double their chromosome number; that cells with excessive chromosome numbers may be cytogenetically unstable, both losing chromosomes randomly during subsequent cell divisions, and often developing structural abnormalities in the chromosomes that are retained; and that some structural chromosome abnormalities may activate growth-promoting genes. The sequence of tetraploidization with chromosome loss can occur repeatedly in a given tumor. The available evidence supporting the model is reviewed. A computer simulation system that embodies these concepts is described and the model is used to generate distributions of chromosome number/cell under various simulated conditions and in a variety of simulated biological settings. A simulation of the time course of changes in chromosome number per cell that accompany the spontaneous neoplastic transformation of mouse fibroblasts in vitro is described. The best fit to the data was obtained when provision was made for the activation of at least two growth-promoting genes. The conditions for generating discrete aneuploid peaks in cytogenetic and flow cytometric studies were explored; our modeling studies suggest that the activation of a growth promoting gene is required in order to produce a discrete aneuploid peak. Our modeling studies suggest that the overrepresentation of individual oncogene-bearing chromosomes in aneuploid cell lines may require the activation of gene dose-dependent growth-promoting genes and is not likely to occur in cell lines in which at least two copies of each normal chromosome are required for cell survival. Overall, the results obtained using the model are consistent with a wide variety of flow cytometric and cytogenetic studies in human solid tumors.

Renal transplantation one week after conception.

A successful 38-week pregnancy is reported following renal transplantation approximately 1 week after conception. The patient was treated with quadruple sequential induction therapy, maintenance immunosuppression, and routine posttransplantation care, including ganciclovir treatment for a symptomatic cytomegalovirus infection during the pregnancy and 3 months after delivery. No decline in renal function was noted. The mother and child remain healthy at 18 months. This case demonstrates the ability of renal transplant patients to maintain renal function throughout pregnancy and the lack of deleterious effects upon the child during gestation and at up to 18 months after birth, despite significant immunosuppression, including antithymocyte globulin induction therapy, and infectious complications of the mother's renal transplantation.

Design and performance of a small-animal imaging system using synthetic collimation.

This work outlines the design and construction of a single-photon emission computed tomography imaging system based on the concept of synthetic collimation. A focused multi-pinhole collimator is constructed using rapid-prototyping and casting techniques. The collimator projects the centre of the field of view (FOV) through 46 pinholes when the detector is adjacent to the collimator, with the number reducing towards the edge of the FOV. The detector is then moved further from the collimator to increase the magnification of the system. The object distance remains constant, and each new detector distance is a new system configuration. The level of overlap of the pinhole projections increases as the system magnification increases, but the number of projections subtended by the detector is reduced. There is no rotation in the system; a single tomographic angle is used in each system configuration. Image reconstruction is performed using maximum-likelihood expectation-maximization and an experimentally measured system matrix. The system matrix is measured for each configuration on a coarse grid, using a point source. The pinholes are individually identified and tracked, and a Gaussian fit is made to each projection. The coefficients of these fits are used to interpolate the system matrix. The system is validated experimentally with a hot-rod phantom. The Fourier crosstalk matrix is also measured to provide an estimate of the average spatial resolution along each axis over the entire FOV. The 3D synthetic-collimator image is formed by estimating the activity distribution within the FOV and summing the activities in the voxels along the axis perpendicular to the collimator face.

Accumbens Homer2-mediated signaling: a factor contributing to mouse strain differences in alcohol drinking?

Alcohol-induced increases in nucleus accumbens glutamate actively regulate alcohol consumption, and the alcohol responsiveness of corticoaccumbens glutamate systems relates to genetic variance in alcohol reward. Here, we extend earlier data for inbred mouse strain differences in accumbens glutamate by examining for differences in basal and alcohol-induced changes in the striatal expression of glutamate-related signaling molecules between inbred C57BL/6J and DBA2/J mice. Repeated alcohol treatment (8 × 2 g/kg) increased the expression of Group1 metabotropic glutamate receptors, the NR2a/b subunits of the N-methyl-D-aspartate receptor, Homer2a/b, as well as the activated forms of protein kinase C (PKC) epsilon and phosphoinositol-3-kinase within ventral, but not dorsal, striatum. Regardless of prior alcohol experience, C57BL/6J mice exhibited higher accumbens levels of mGluR1/5, Homer2a/b, NR2a and activated kinases vs. DBA2/J mice, whereas an alcohol-induced rise in dorsal striatum mGluR1/5 expression was observed only in C57BL/6J mice. We next employed virus-mediated gene transfer approaches to ascertain the functional relevance of the observed strain difference in accumbens Homer2 expression for B6/D2 differences in alcohol-induced glutamate sensitization, as well as alcohol preference/intake. Manipulating nucleus accumbens shell Homer2b expression actively regulated these measures in C57BL/6J mice, whereas DBA2/J mice were relatively insensitive to the neurochemical and behavioral effects of virus-mediated changes in Homer2 expression. These data support the over-arching hypothesis that augmented accumbens Homer2-mediated glutamate signaling may be an endophenotype related to genetic variance in alcohol consumption. If relevant to humans, such data pose polymorphisms affecting glutamate receptor/Homer2 signaling in the etiology of alcoholism.

Prophylactic versus preemptive oral valganciclovir for the management of cytomegalovirus infection in adult renal transplant recipients.

Prophylaxis reduces cytomegalovirus (CMV) disease, but is associated with increased costs and risks for side effects, viral resistance and late onset CMV disease. Preemptive therapy avoids drug costs but requires frequent monitoring and may not prevent complications of asymptomatic CMV replication. Kidney transplant recipients at risk for CMV (D+/R-, D+/R+, D-/R+) were randomized to prophylaxis (valganciclovir 900 mg q.d. for 100 days, n=49) or preemptive therapy (900 mg b.i.d. for 21 days, n=49) for CMV DNAemia (CMV DNA level>2000 copies/mL in >or=1 whole blood specimens by quantitative PCR) assessed weekly for 16 weeks and at 5, 6, 9 and 12 months. More patients in the preemptive group, 29 (59%) than in the prophylaxis group, 14 (29%) developed CMV DNAemia, p=0.004. Late onset of CMV DNAemia (>100 days after transplant) occurred in 11 (24%) randomized to prophylaxis, and none randomized to preemptive therapy. Symptomatic infection occurred in five patients, four (3 D+/R- and 1 D+/R+) in the prophylactic group and one (D+/R-) in the preemptive group. Peak CMV levels were highest in the D+/R- patients. Both strategies were effective in preventing symptomatic CMV. Overall costs were similar and insensitive to wide fluctuations in costs of either monitoring or drug.

Endoscopic carpal tunnel release using the subligamentous two-portal technique.

The surgical treatment for entrapment neuropathy of the median nerve in the carpal tunnel is varied. Recent publications have demonstrated a closed, endoscopic method for release of the carpal tunnel using a two-portal technique. In this report, a surgical technique for performing a subligamentous modification of this two-portal procedure is discussed, and the special tools used in this approach are demonstrated. In the early portion of this series of 75 symptomatic hands in 65 patients, 8% (6/75 hands) developed transient postoperative ulnar nerve neuropraxia. One patient experienced an iatrogenic laceration of the median nerve. Following the institution of the subligamentous approach, no postoperative nerve complications occurred. The majority of the patients experienced a significant decrease in their median nerve symptoms. This procedure has been found to be safe and is equally as effective as open surgical treatment of patients with carpal tunnel syndrome.

Cellular transformation by adenovirus type 5 is influenced by the viral DNA polymerase.

Early region 2B (E2B) of the group C adenoviruses encodes a number of proteins, including the 140-kilodalton DNA polymerase, which plays a role in the initiation of viral DNA replication. Temperature-sensitive (ts) mutants with mutations mapping to E2B are conditionally defective for both DNA replication in human cells and transformation of rat cells. Nucleotide sequence analysis shows that the E2B mutant ts36 possesses a single point mutation specific to the viral DNA polymerase; this transition of a C to a T at position 7623 changes leucine residue 249 in the polymerase to a phenylalanine. A wild-type (ts+) revertant possesses a codon specifying the original leucine at position 249. Phenotypic analysis of revertant and wild-type viruses derived by marker rescue from ts36 shows that these variants are wild type for both viral DNA replication and transformation. Thus, the single point mutation in the polymerase gene of ts36 is responsible for both defects.

18-substituted steroids--9[1]. Studies on the stability of aldosterone in dilute alkali.

In the presence of dilute alkali at room temperature aldosterone undergoes rearrangement to form an isomeric compound identified as 11 beta, 18;18,21-diepoxy-20,21-dihydroxypregn-4-en-3-one. The presence of dissolved oxygen causes simultaneous degradation to form 11 beta, 18-epoxy-18-hydroxy-3-oxo-17(beta H)-androst-4-ene-17 alpha-carboxylic acid. Under similar alkaline conditions at reflux temperature in aqueous methanol aldosterone undergoes rearrangement to form (20S)-20,21-dihydroxy-3-oxo-pregn-4-eno-18,11 beta-lactone accompanied by an intramolecular aldol type condensation to form 11 beta, 21-dihydroxy-18,21-cyclopregna-4,18(21)-diene-3,20-dione. Identical products were formed from 17-isoaldosterone. The mechanisms of these reactions are discussed.

Assessment of side effects in patients with chronic hepatitis C receiving combination therapy.

The purpose of this descriptive study is to review the adverse effects of combination therapy, interferon alfa-2b and ribavirin, in a sample of patients with chronic hepatitis C who were part of a larger multi-center trial (Bonkovsky, 1999). The sample (n = 13) was drawn from one hepatology practice in the northeastern United States. This secondary analysis reported sums, frequencies, means, and standard deviations for the sample. Patients who received 600 mg ribavirin per day in addition to interferon alfa-2B (Arm A) showed a mean of 9.5 different types of side effects. Patients who received 1000-1200 mg of ribavirin in addition to interferon alfa-2B (Arm B) had a mean of 8.6 different types of side effects. The highest-ranking side effects were fatigue (92%), insomnia (85%), alopecia (69%), and arthralgia (62%). A secondary analysis of the entire data set is required before confidence can be placed in the findings. Implications for nursing practice include the need for creative strategies to reduce the effects of fatigue, insomnia, alopecia, and arthralgia in patients with chronic hepatitis C.

Overexpression of protein kinase C-zeta isoform increases cyclooxygenase-2 and inducible nitric oxide synthase.

Cyclooxygenase (COX) catalyzes the formation of prostaglandins from arachidonic acid. Nitric oxide synthase catalyzes the production of nitric oxide, a short-lived messenger molecule involved in many diverse cellular processes. Both of these enzymes have inducible forms [COX-2 and inducible nitric oxide synthase (iNOS), respectively] that respond to environmental stresses, chemicals, and extracellular ligands such as interleukin-1, epidermal growth factor, and platelet-derived growth factor. The precise cascade of intracellular events that leads to the expression of either COX-2 or iNOS is not known. Protein kinase C (PKC) is a family of 11 serine-threonine kinases conserved throughout eukaryotic species that transduce a wide variety of signals critical for cellular functions. Using a retroviral vector to overexpress the zeta-isoform of PKC in rat mesangial cells, we demonstrate markedly increased COX-2, prostaglandin E2 (PGE2), iNOS, and altered cellular morphology compared with mesangial cells expressing a control retroviral vector and untransfected mesangial cells. NIH/3T3 cells overexpressing PKC-zeta showed no change in morphology, PGE2 production, COX-2 expression, or iNOS expression at basal conditions. This suggests a role for PKC-zeta in the expression of these enzymes in mesangial cells.

Hand and wrist involvement in calcium pyrophosphate dihydrate crystal deposition disease.

The clinical records and hand and wrist radiographs of 51 patients with calcium pyrophosphate dihydrate crystal deposition disease have been analyzed, and symptomatology and radiologic abnormalities have been correlated. Characteristic roentgenographic features included cartilage and synovial calcification and arthropathy of the metacarpophalangeal joints and the radiocarpal compartment of the wrist, including scapholunate dissociation. Clinical-radiologic correlation revealed many asymptomatic patients with calcification and arthropathy and many symptomatic patients with normal radiographs. Thorough radiologic evaluation may reveal many patients with this disorder before the onset of clinical symptoms.

Interleukin-1beta-induced cyclooxygenase-2 expression requires activation of both c-Jun NH2-terminal kinase and p38 MAPK signal pathways in rat renal mesangial cells.

The inflammatory cytokine interleukin-1beta (IL-1beta) induces cyclooxygenase-2 (Cox-2) expression with a concomitant release of prostaglandins from glomerular mesangial cells. We reported previously that IL-1beta rapidly activates the c-Jun NH2-terminal/stress-activated protein kinases (JNK/SAPK) and p38 mitogen-activated protein kinase (MAPK) and also induces Cox-2 expression and prostaglandin E2 (PGE2) production. The current study demonstrates that overexpression of the dominant negative form of JNK1 or p54 JNK2/SAPKbeta reduces Cox-2 expression and PGE2 production stimulated by IL-1beta. Similarly, overexpression of the kinase-dead form of p38 MAPK also inhibits IL-1beta-induced Cox-2 expression and PGE2 production. These results suggest that activation of both JNK/SAPK and p38 MAPK is required for Cox-2 expression after IL-1beta activation. Furthermore, our experiments confirm that IL-1beta activates MAP kinase kinase-4 (MKK4)/SEK1, MKK3, and MKK6 in renal mesangial cells. Overexpression of the dominant negative form of MKK4/SEK1 decreases IL-1beta- induced Cox-2 expression with inhibition of both JNK/SAPK and p38 MAPK phosphorylation. Overexpression of the kinase-dead form of MKK3 or MKK6 demonstrated that either of these two mutant kinases inhibited IL-1beta-induced p38 MAPK phosphorylation and Cox-2 expression but not JNK/SAPK phosphorylation and activation. This study suggests that the activation of both JNK/SAPK and p38 MAPK signaling cascades is required for IL-1beta-induced Cox-2 expression and PGE2 synthesis.

The extensor retinaculum of the wrist.

The dorsal retinaculum of the wrist consists of two layers: the supratendinous and the infratendinous. The infratendinous layer is limited to an area deep to the ulnar three compartments. There are six compartments for the tendons dorsal to the wrist separated by six longitudinal vertical septa. Each septum originates from the supratendinous retinaculum and inserts onto the radius. The sixth compartment for the extensor carpi ulnaris is complex. The tendon of the extensor carpi ulnaris is enclosed in an independent fibrous tunnel formed by the supratendinous retinaculum superiorly, the infratendinous retinaculum inferiorly, the sixth septum laterally, and the ulnar insertion of the retinaculum reinforced by longitudinal fibers called the "linea jugata" medially. Our findings support the concept of an adaptable dynamic collateral ligament system rather than the traditional radial and ulnar collateral ligaments.

Common variable immunodeficiency in a renal transplant patient with severe recurrent bacterial infection: a case report and review of the literature.

The second reported case of common variable immunodeficiency (acquired agammaglobulinemia) after renal transplantation is presented. Agammaglobulinemia presumably resulted from long-standing immunosuppression. This case and our review of the literature indicate that agammaglobulinemia is a rare event after transplantation but can be treated successfully with intravenous immunoglobulin. Additionally, hypogammaglobulinemia occurs frequently after transplantation and should be monitored and treated in appropriate clinical situations. The treatment of our patient with intravenous immunoglobulin also suggests that patients with common variable immunodeficiency can undergo renal transplantation.

The Barnes-Jewish Hospital/Washington University Renal Transplant Program: comparison of two eras 1991-1994 and 1995-2000.

The first cadaveric transplant at Barnes-Jewish Hospital/Washington University was performed in 1963, the first living related transplant in 1965, and the first living unrelated transplant in 1983. Changes in the renal transplant program initiated in 1993 and 1994 resulted in many improvements over the past decade. Our comparison of 2 modern eras of transplant, 1991-1994 and 1995-2000, showed the following: 1. No significant differences in patient and donor characteristics. 2. Trends toward greater use of living donors (p = 0.07), older cadaveric donors (p = 0.084) and particularly cadaveric donors > 55 years of age (p = 0.09). 3. Decreasing mean CIT: 19.2 hours vs. 14.2 hours (p < 0.001). 4. Decreasing use of donors with CIT > 24 hours: 22% to 3%, (p < 0.001). 5. Decreased rate of DGF: 13% vs. 8% (p = 0.044). 6. Decreased rate of symptomatic CMV: 35% vs. 14% (p < 0.001). 7. Decreased rate of PTLD: 3.5% vs. 0.5% (p = 0.004). 8. Decreased one-year rate of acute rejection: 41% vs. 15% (p < 0.001). 9. Current one-year rate of acute rejection < 8%. 10. Decreased length of initial hospital stay: 12.7 days to 8.0 days (p < 0.001). 11. Decreased length of hospital in the first year after transplant: 10.6 days vs. 6.4 days (p < 0.001). 12. There were no improvements in patient and graft survival at one and 3 years. a. one-year patient survival rates: 95% vs. 96%. b. 3-year patient survival rates: 90% vs. 90%. c. one-year death-censored graft survival rates: 91% vs. 94%. d. 3-year death-censored graft survival rates: 87% vs. 88%.