Tuberculosis mimicking cervical carcinoma--case report.

Tuberculosis is a chronic bacterial infection that primarily results in pulmonary disease. Although there are several reported cases of extra-pulmonary tuberculosis, very few reports have described this disease in the female genital tract. We present a case involving a 67-year-old woman who presented with vaginal discharge, abdominal discomfort, and a pelvic mass in 2006. Clinically, cervical carcinoma was suspected, but pathologic diagnosis eventually revealed tuberculosis of the cervix. Tuberculosis is associated with a significant inflammatory reaction, which may mimic a gynecologic malignancy on exam or with diagnostic imaging. Despite the rare incidence, tuberculosis of the cervix should be considered in the differential diagnosis when cervical carcinoma is initially suspected.

The evaluation of mixtures of yeast and potato extracts in growth media for biomass production of lactic cultures.

The effectiveness of yeast extracts (YE) and potato extracts (PE) to promote growth of seven lactic cultures was evaluated by automated spectrophotometry (AS). Two aspects of the growth curve were analysed: (1) maximum biomass obtained (using ODmax) and (2) highest specific growth rate mu(max)) Eleven lots from the same PE-manufacturing process were examined for lot-to-lot variability. The ODmax values of three of the seven strains were significantly affected by lot source, but mu(max) was not significantly affected. The growth of bacteria was systematically lower in base medium containing 100% PE than in base medium containing 100% YE for both ODmax or mu(max) data, which could be related to the lower content in nitrogen-based compounds in PE. In AS assays, highest OD values for Lactobacillus casei EQ28, Lactobacillus rhamnosus R-011, Lactobacillus plantarum EQ12, and Streptococcus thermophilus R-083 were obtained with a mixture of PE and YE. Fermentations (2 L) were also carried out to determine the accuracy of AS to predict biomass levels obtained under fermentation trials. In these fermentations, replacement of 50% YE with PE was shown to enable good growth of S. thermophilus. With L. rhamnosus R-011, a high correlation (R2 = 0.95) was found between ODmax data obtained in the AS assays and that of the 2-L bioreactor when the same growth medium was used for both series of fermentations. However, AS was not as efficient when industrial media were used for the bioreactor assays. The relationship was still good for ODmax between AS data and that of the bioreactor data with L. rhamnosus R-011 in industrial LBS medium (R2 = 0.87), but was very poor with the S. thermophilus R-083 on Rosell #43 industrial medium (R2 = 0.33). Since PE cost 40% less than YE, there are strong economic advantages in considering such a partial replacement of YE by PE.

Drinking behavior and motivation for treatment among alcohol-dependent liver transplant candidates.

Alcohol misuse is the second most common indication for liver transplantation in the United States. Our post-transplant alcoholism treatment trial suggested that current interventions might not be transferable to liver transplantees. We sought to identify differences between patients awaiting liver transplantation and alcoholics entering treatment without severe liver disease. Thirty transplant patients were compared to thirty naltrexone study patients on medical status, alcohol and drug use, alcohol craving, motivation for treatment, psychiatric symptoms, and psychosocial problems. Lifetime alcohol consumption was greater for transplant patients compared to naltrexone patients. In contrast to the naltrexone group, transplant patients denied craving for alcohol and showed little motivation for alcoholism treatment. Groups did not differ on other psychosocial measures. Liver transplant patients differ from patients in alcoholism treatment trials on measures of alcohol consumption, alcohol craving and motivation for treatment. Alcoholism interventions should accommodate their medical condition and boost motivation for continued abstinence.

A pilot test of motivational interviewing groups for dually diagnosed inpatients.

Motivational interviewing is a brief treatment approach designed to produce rapid, internally motivated change in addictive behaviors. Motivational interviewing shows promise for engaging clients with dual psychiatric and psychoactive substance use diagnoses in treatment. While initially developed as an individual treatment approach, key motivational enhancement principles may be applied to structured group interventions to facilitate its introduction to inpatient dual-diagnosis treatment. We describe how we developed and pilot-tested a motivational interviewing group for dually diagnosed inpatients, and illustrate successes and pitfalls in clinical implementation. Group participants were readily engaged by the entertaining format and often provocative content, and appeared to benefit from exploration of their ambivalence regarding change. Directions for further development and evaluation are proposed.

Alcoholism treatment after liver transplantation: lessons learned from a clinical trial that failed.

Alcoholic liver disease is the second most common indication for liver transplantation in the United States. The lack of alcoholism treatment studies led us to study motivational enhancement therapy (MET) plus naltrexone after transplant. The authors could not complete this study. Sixty alcoholic patients were to receive MET plus naltrexone or placebo for 6 months. Fifty men and 5 women were screened. Nine died and 15 were not approached. Of 31 approached, 20 were ineligible, 11 refused, and 5 entered but dropped out before completion. Barriers to posttransplant alcoholism included infirmity, intensive medical management, and denial for alcoholism treatment. Because 30%-50% of alcoholic patients drink after transplant, the authors suggest using MET alone pretransplant.

Interpreting the significance of drinking by alcohol-dependent liver transplant patients: fostering candor is the key to recovery.

Few studies have examined the value of treating alcohol addiction either before or after liver transplantation. Nevertheless, most liver transplant programs and many insurance companies require 6 months to 1 year of abstinence from alcohol as a condition of eligibility for liver transplantation (the 6-month rule). We believe there are potentially harsh clinical consequences to the implementation of this rule. For example, the natural history of alcohol use disorders often involves brief fallbacks to drinking ("slips"), but when alcoholic liver transplant candidates slip, most are removed from consideration for transplantation or are required to accrue another 6 months of sobriety. Because there is no alternative treatment to liver transplantation for most patients with end-stage liver disease, the 6-month rule could be lethal in some circumstances. In this review, we survey the literature concerning the ability of the 6-month rule to predict drinking by alcoholic patients who undergo liver transplantation and examine its impact on the health consequences of drinking before and after liver transplantation. We believe that fostering candor between the alcoholic patient and the transplant team is the key to recovery from alcoholism. We conclude that it is unethical to force alcoholic liver patients who have resumed alcohol use while waiting for or after transplantation to choose between hiding their drinking to remain suitable candidates for transplantation or risk death by asking for treatment of alcoholism. Consequently, we advocate a flexible approach to clinical decision making for the transplant professional caring for an alcoholic patient who has resumed drinking and provide specific guidelines for patient management.

Endogenous generation of cyanide in neuronal tissue: involvement of a peroxidase system.

In a study of the mechanism by which cyanide is produced in neural tissue, it was hypothesized that nerve cells generate cyanide in a manner similar to that in leukocytes. As in white blood cells, glycine addition enhanced cyanide production in rat pheochromocytoma cells. Because myeloperoxidase catalyses cyanide production in leukocytes, a selective myeloperoxidase inhibitor (aminobenzoic acid hydrazide) was tested and found to inhibit opiate agonist-induced cyanide production in pheochromocytoma cells and also in rat brain. In addition, hydrogen peroxide enhanced cyanide release in pheochromocytoma cells, further suggesting that the process is oxidative in nature. Sonicated rat pheochromocytoma cells did not generate cyanide in response to an agonist acting on surface receptors even though disrupted cells responded to glycine. The mitochondrial fraction from rat brain produced more cyanide in response to glycine than any other fraction. Thus glycine seems to act at an intracellular site to enhance cyanide production and the process seems to involve a peroxidase mechanism similar to that reported for white blood cells.

Immunogenicity of antigens in boiled alginate microspheres.

Vaccine efficacy can be enhanced by delivery of antigens in synthetic microspheres. The process of antigen incorporation into microspheres can expose fragile antigens to damaging conditions, such as high temperatures, and to bacterial contamination. Maintenance of immunogenicity of several antigens and reduction of bacterial load in alginate microspheres following boiling was evaluated. Mice were immunized subcutaneously, initially and again 21 days later, with either non-boiled or boiled microspheres containing ovalbumin (OVA), a culture supernatant vaccine of Pasteurella haemolytica (PHV), or a potassium thiocyanate extract of P. multocida (PTE). Serum samples were obtained prior to immunization and at the time of euthanasia 28 days later. Culture of microspheres showed that boiling completely eliminated aerobic bacterial growth for OVA-containing microspheres, and reduced growth by a factor of 10(4) for PTE microspheres. More bacteria were cultured after boiling than before for PHV microspheres. ELISA performed on serum and intestinal lamina propria explant supernatants showed that immunogenicity of PHV microspheres was not altered by boiling. Boiled OVA microspheres were still able to stimulate a significant serum IgG anti-OVA titer in mice, but boiled PTE microspheres completely lacked immunogenicity. Elispot assays of spleens showed that only PHV microspheres were able to retain immunogenicity after boiling. Results indicate that boiling is not an effective means for reducing the bacterial load of alginate microspheres and that the process is associated with a diminution of vaccine immunogenicity.

Substance-use situations and abstinence predictions in substance abusers with and without personality disorders.

Rates of personality disorders (PDs) in substance abusers are higher than in the general population. Comorbid PDs are believed to complicate the treatment of addicted patients: in addition to having more severe substance-use disorders and life problems, personality-disordered patients may use substances differently than their peers without Axis II diagnoses. In a sample of 339 adults receiving inpatient treatment for alcohol or drug abuse/dependence, 71.7% received Axis II diagnoses, and they presented a more severe clinical picture. They also had more self-reported "impulsive" substance use and use of drugs or alcohol in positive situations. Different groups of personality-disordered patients had different patterns of self-efficacy for abstinence for hypothetical future situations.

The La protein in Schizosaccharomyces pombe: a conserved yet dispensable phosphoprotein that functions in tRNA maturation.

Most RNA polymerase III transcripts are bound immediately after synthesis by an abundant nuclear phosphoprotein known as the La autoantigen. Experiments performed in the budding yeast Saccharomyces cerevisiae have revealed that binding of the La protein to tRNA precursors is required for the endonucleolytic maturation of the 3' terminus of many tRNAs. In the absence of this protein, the 3' ends of these tRNAs are trimmed by exonucleases (Yoo CJ, Wolin SL, 1997, Cell 89:393-402). Here we report the characterization of the La protein in the fission yeast Schizosaccharomyces pombe. As was described for budding yeast, S. pombe cells lacking the La protein are viable and exhibit alterations in the pathway of pre-tRNA maturation. Introduction of either the human, S. cerevisiae, or S. pombe La protein into these cells restores the detected pattern of tRNA processing intermediates to that of wild-type cells. By performing immunoprecipitations from cells that were metabolically labeled with 32P-orthophosphate, we demonstrate that the S. pombe and S. cerevisiae La proteins, like the human La protein, are phosphorylated in vivo. Thus, although the La protein is dispensable for growth in these yeasts, both the structure of the protein and its function in pre-tRNA maturation have been highly conserved throughout evolution.

A misfolded form of 5S rRNA is complexed with the Ro and La autoantigens.

In both vertebrate and invertebrate cells, the 60-kDa Ro autoantigen is bound to small cytoplasmic RNAs known as Y RNAs. In Xenopus oocytes, the 60-kDa Ro protein is also complexed with a class of 5S rRNA precursors that contain internal mutations. Because these 5S rRNA precursors are processed inefficiently and degraded eventually, the Ro protein may function in a quality control pathway for 5S rRNA biosynthesis. We have investigated the sequence and secondary structure determinants in the mutant 5S rRNAs that confer binding by the 60-kDa Ro protein. The mutant 5S rRNAs fold to form an alternative helix that is required for recognition by the 60-kDa Ro protein. Mutations that disrupt the alternative helix eliminate Ro protein binding, whereas compensatory changes that restore the helix are bound efficiently by the Ro protein. When the structure of the mutant RNA was probed using dimethylsulfate and oligonucleotide-directed RNase H cleavage, the results were consistent with the formation of the alternative structure. The La protein, which is also complexed with the mutant 5S rRNA precursors, protects similar sequences from nuclease digestion as does the 60-kDa Ro protein. Thus, the binding sites for these two proteins are either nearby on the RNA, or the two proteins may be complexed through protein-protein interactions. When the human Ro protein is expressed in the yeast Saccharomyces cerevisiae, the protein binds wild-type 5S rRNA precursors, suggesting that a population of wild-type precursors also folds into the alternative structure.

Insulin receptor substrate 1 rescues insulin action in CHO cells expressing mutant insulin receptors that lack a juxtamembrane NPXY motif.

Insulin signals are mediated through tyrosine phosphorylation of specific proteins such as insulin receptor substrate 1 (IRS-1) and Shc by the activated insulin receptor (IR). Phosphorylation of both proteins is nearly abolished by an alanine substitution at Tyr-960 (A960) in the beta-subunit of the receptor. However, overexpression of IRS-1 in CHO cells expressing the mutant receptor (A960 cells) restored sufficient tyrosine phosphorylation of IRS-1 to rescue IRS-1/Grb-2 binding and phosphatidylinositol 3' kinase activation during insulin stimulation. Shc tyrosine phosphorylation and its binding to Grb-2 were impaired in the A960 cells and were unaffected by overexpression of IRS-1. Although overexpression of IRS-1 increased IRS-1 binding to Grb-2, ERK-1/ERK-2 activation was not rescued. These data suggest that signaling molecules other than IRS-1, perhaps including Shc, are critical for insulin stimulation of p21ras. Interestingly, overexpression of IRS-1 in the A960 cells restored insulin-stimulated mitogenesis and partially restored insulin stimulation of glycogen synthesis. Thus, IRS-1 tyrosine phosphorylation is sufficient to increase the mitogenic response to insulin, whereas insulin stimulation of glycogen synthesis appears to involve other factors. Moreover, IRS-1 phosphorylation is either not sufficient or not involved in insulin stimulation of ERK.

Caenorhabditis elegans embryos contain only one major species of Ro RNP.

In virtually all vertebrate cells, Ro RNPs consist of the 60-kDa Ro autoantigen bound to one of several small cytoplasmic RNA molecules known as Y RNAs. Because the 60-kDa Ro autoantigen is also found complexed with defective precursors of 5S rRNA in Xenopus oocytes, we have proposed that this protein functions in a quality control, or discard pathway, for 5S RNA biosynthesis (O'Brien CA, Wolin SL, 1994, Genes & Dev 8:2891-2903). The role of the Y RNAs in this pathway is unknown. To begin a genetic analysis of Ro RNP function, we have characterized these particles in the nematode Caenorhabditis elegans. The C. elegans Ro protein is 12 kDa larger than the vertebrate protein; the larger size is due in part to an N-terminal extension and to two insertions in the RNA recognition motif. In contrast to all previously described vertebrate species, the Ro protein appears bound to a single Y RNA in C. elegans. Similar to vertebrate Y RNAs, the C. elegans Y RNA can be folded to form a pyrimidine-rich internal loop and a long stem in which the 5' and 3' ends are base paired. Within the stem is a conserved bulged helix that is proposed to be the binding site of the Ro protein. Interestingly, although the human protein can bind the nematode Y RNA, the C. elegans protein does not bind human Y RNAs. This is the first description of Ro RNPs in an invertebrate species.

Regulation of phosphatidylinositol 3'-kinase by tyrosyl phosphoproteins. Full activation requires occupancy of both SH2 domains in the 85-kDa regulatory subunit.

Phosphatidylinositol 3'-kinase (PI 3'-kinase) is activated in insulin-stimulated cells by the binding of the SH2 domains in its 85-kDa regulatory subunit to insulin receptor substrate-1 (IRS-1). We have previously shown that both tyrosyl-phosphorylated IRS-1 and mono-phosphopeptides containing a single YXXM motif activate PI 3'-kinase in vitro. However, activation by the monophosphopeptides was significantly less potent than activation by the multiply phosphorylated IRS-1. We now show that the increased potency of PI 3'-kinase activation by IRS-1 relative to phosphopeptide is not due to tertiary structural features IRS-1, as PI 3'-kinase is activated normally by denatured, reduced, and carboxymethylated IRS-1. Furthermore, activation of PI 3'-kinase by bis-phosphorylated peptides containing two YXXM motifs is 100-fold more potent than the corresponding mono-phosphopeptides and similar to activation by IRS-1. These data suggest that tyrosyl-phosphorylated IRS-1 or bis-phosphorylated peptides bind simultaneously to both SH2 domains of p85. However, these data cannot differentiate between an activation mechanism that requires two-site occupancy for maximal activity as opposed to one in which bivalent binding enhances the occupancy of a single activating site. To distinguish between these possibilities, we produced recombinant PI 3'-kinase containing either wild-type p85 or p85 mutated in its N-terminal, C-terminal, or both SH2 domains. We find that mutation of either SH2 domains significantly reduced phosphopeptide binding and decreased PI 3'-kinase activation by 50%, whereas mutation of both SH2 domains completely blocked binding and activation. These data provide the first direct evidence that full activation of PI 3'-kinase by tyrosylphosphorylated proteins requires occupancy of both SH2 domains in p85.

Direct activation of the phosphatidylinositol 3'-kinase by the insulin receptor.

We have previously shown that phosphatidylinositol (PtdIns) 3'-kinase is activated by the binding of proteins or peptides containing the phosphorylated motif Y(P)XXM. In the present study, we examine interactions between PtdIns 3'-kinase and the human insulin receptor, which contains a C-terminal phosphorylation site in the sequence Y1322THM. Partially purified insulin receptors bound tightly to bacterial fusion proteins containing the N- or C-terminal SH2 domains from PtdIns 3'-kinase regulatory subunit (p85). In contrast, a mutant insulin receptor, truncated by 43 amino acids at the C terminus (IR delta CT), bound poorly to the SH2 domains; these mutant receptors have normal kinase activity but lack the Y1322THM motif. Similarly, incubation with wild-type receptors increased the activity of immunopurified PtdIns 3'-kinase, whereas incubation with IR delta CT receptors did not affect PtdIns 3'-kinase activity. Activation of PtdIns 3'-kinase by the wild-type receptor was mimicked by a tyrosyl phosphopeptide derived from the insulin receptor C terminus and containing the Y1322THM motif; non-phosphorylated peptide did not affect activity. Thus, the insulin receptor C terminus activates PtdIns 3'-kinase in vitro by binding to the SH2 domains of the 85-kDa regulatory subunit. These data support the hypothesis that binding of tyrosyl-phosphorylated receptors to p85 SH2 domains is a general mechanism for PtdIns 3'-kinase activation, and they suggest that direct interactions between the insulin receptor and PtdIns 3'-kinase may provide an alternative pathway for the activation of this enzyme by insulin.

Identifying young adult substance abusers: the Rutgers Collegiate Substance Abuse Screening Test.

Substance abuse is a serious problem in the young adult population, yet there exists a lack of reliable screening measures for use in identifying problem users in this age group. The Rutgers Collegiate Substance Abuse Screening Test (RCSAST) is a 25-item, true/false questionnaire that was created to provide a reliable means of identifying young adult substance abusers. Three groups completed the RCSAST: a clinical sample of 84 young adult, problem substance users; a group of 33 young adults who were referred to an assistance program but were judged not to have a substance use problem; and a control sample of 87 young adult, nonproblem substance users. The RCSAST correctly classified 94% of the clinical subjects as problem users, and 89% of the control subjects as nonproblem users. The difference between the average total scores for the two groups was highly significant. In addition, the RCSAST was able to distinguish between problem and nonproblem users within the sample of subjects who were referred for evaluation. The findings support the use of the RCSAST in identifying young adult substance abusers.

Metallic ballistic fragments: MR imaging safety and artifacts.

The ferromagnetism of various bullets and shotgun pellets was tested in vitro. Magnetic deflection showed that four of 21 metallic specimens tested (all bullets) demonstrated marked ferromagnetism. Three of these four were made outside the United States; two of the four were known to contain steel, and the other two were reportedly either copper or copper-nickel-jacketed lead bullets (indicating that the ferromagnetism was due to impurities in the bullet jackets or cores). Ferromagnetic bullets readily rotated within a gelatin phantom in response to magnetic torque. Nonferromagnetic bullets and pellets demonstrated only mild to moderate metal artifact during spin-echo and gradient-echo magnetic resonance (MR) imaging. However, all four of the ferromagnetic bullets produced severe MR artifacts and image distortion. MR studies of seven patients with retained bullets, pellets, or shrapnel were reviewed. In six of the seven, only mild MR artifacts were seen. Only intracranial shrapnel (presumably steel) in one patient created significant artifact. All seven patients with retained bullets and shotgun pellets were imaged safely with MR. However, caution should be exercised with MR imaging in the presence of metallic foreign bodies, particularly if they are located near vital neural, vascular, or soft-tissue structures.

Comparison of the Effects of the Anderson Knee Stabler and McDavid Knee Guard on the Kinematics of the Lower Extremity during Gait*.

* This study was completed in partial fulfillment of Ms. Van Horn's master's degree, University of North Carolina at Chapel Hill. The purpose of this study was to compare gait patterns among subjects wearing Anderson Knee Stabler braces, McDavid Knee Guards, and no braces. Fifteen male subjects were filmed while running on a treadmill at 4 mph and 8 mph without a brace, with an Anderson Knee Stabler, and with a McDavid Knee Guard. Fourteen gait variables were measured for each brace and speed condition. Analysis of the variables with multivariate ANOVA indicated that there was an increase in hip and knee flexion and knee angular velocity with and without braces at 8 mph as compared to 4 mph, a decrease in knee extension when either brace was worn, and minimal gait pattern differences with the Anderson Knee Stabler as compared with the McDavid Knee Guard (all results p < 0.05). The results of this study demonstrate that no clear superiority exists between the braces' effect on the gait characteristics measured. Therefore, other parameters should be considered when making brace selections.J Orthop Sports Phys Ther 1988;9(7):254-260.

Human parvovirus-associated red cell aplasia in the absence of underlying hemolytic anemia.

Human parvovirus (HPV) infection has recently been implicated as the cause of aplastic crisis in patients with hemolytic anemias such as congenital spherocytosis and sickle cell anemia. The virus causes a transient red cell aplasia which, in patients with a shortened red cell life span, is manifested as a rapid worsening of the anemia and an absence of peripheral reticulocytosis. Recovery is associated with the presence of giant pronormoblasts in the bone marrow, and several days later, a brisk peripheral reticulocytosis. In normal subjects, HPV causes erythema infectiosum (fifth disease) but is not associated with symptomatic anemia, probably because of the duration of the normal red blood cell life span. A case of HPV infection producing severe anemia in an immunocompromised patient without an underlying hemolytic anemia is presented here. Infection in this patient, a 3-year-old boy with acute lymphoblastic leukemia in remission, may have been prolonged by immunosuppression, leading over a 4-week period to a severe anemia. The immunosuppressed appear to be another group of patients at risk of developing symptomatic anemia when infected by HPV.