A Comparison of Trabeculectomy Surgery Outcomes With Mitomycin-C Applied by Intra-Tenon Injection Versus Sponge.

PURPOSE: To compare the outcomes of mitomycin-C (MMC) delivered by intra-Tenon injection vs sponge application during trabeculectomy surgery. METHODS: We retrospectively reviewed 566 patients with primary and secondary glaucoma diagnoses who received trabeculectomy surgery with MMC in an academic medical center. Exclusion criteria were age less than 18 years, no light perception vision, combined surgery, previous glaucoma incisional surgery, intraoperative 5-fluorouracil, or follow-up <1 month. Subjects were divided into 2 cohorts: MMC delivered by sponge application or by intra-Tenon injection. Main outcome measures were postoperative intraocular pressure (IOP) level and secondary measures were survival rate for IOP control, glaucoma medication use, complication rate, and vision. RESULTS: After inclusion/exclusion criteria, 316 eyes were available for analysis; 131 eyes had MMC delivered via sponge and 185 eyes via injection. Mean postoperative IOP was not significantly different between treatment groups but change in IOP from baseline was lower in the sponge vs the injection group 24 months after surgery (P = .038). The MMC sponge group had significantly more tense, vascularized, or encapsulated blebs as a late complication (P = .046). Time to failure for postoperative IOP control was not significantly different between MMC treatment groups, but older patient age and limbus-based conjunctival incision were associated with significantly longer time to fail. CONCLUSIONS: The application of MMC by injection was similar to application by sponge in lowering IOP in patients with glaucoma and the safety of both techniques appears to be comparable. Limbus-based conjunctival incision had longer time to failure for postoperative IOP control vs fornix-based incision. NOTE: Publication of this article is sponsored by the American Ophthalmological Society.

Quality of life and everyday activities in patients with primary biliary cirrhosis.

UNLABELLED: Primary biliary cirrhosis (PBC) is generally a slowly progressive disease that may lead to cirrhosis and liver failure. However, patients with PBC often suffer from a variety of symptoms long before the development of cirrhosis that include issues of daily living that have an impact on their work environment and their individual quality of life. We therefore examined multiple parameters by taking advantage of the database of our cohort of 1032 patients with PBC and 1041 matched controls. The data were obtained from patients from 23 tertiary referral centers throughout the United States and from rigorously matched controls by age, sex, ethnicity, and random-digit dialing. The data showed that patients with PBC were more likely than controls to have significant articular symptoms, a reduced ability to perform household chores, and the need for help with routine activities. Patients with PBC rated their overall activity similar or superior to that of controls; however, more of them reported limitations in their ability to carry out activities at work or at home and difficulties in everyday activities. PBC cases also more frequently reported limitations in participating in certain sports or exercises and pursuing various hobbies; however, they did not report significant limitations in social activities. In a multivariable analysis, household income, a diagnosis of systemic lupus erythematosus, limitations in work activities, a reduction in work secondary to disability, and church attendance were independently increased in PBC cases with respect to controls. CONCLUSION: Our data indicate that the quality of life of patients with PBC in the United States is generally well preserved. Nevertheless, patients with PBC suffer significantly more than controls from a variety of symptoms that are beyond the immediate impact of liver failure and affect their lifestyle, personal relationships, and work activities.

X monosomy in female systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease, predominantly occurring in women of childbearing age. SLE, like several other autoimmune diseases, is characterized by a striking female predominance and, although sex hormone influences have been suggested as an explanation for this phenomenon, definitive data are still unavailable. Our group recently reported an increased X monosomy in lymphocytes of women, affected with primary biliary cirrhosis (PBC), systemic sclerosis (SSc), and autoimmune thyroiditis (AITD) in comparison to healthy women, thus suggesting the involvement of this chromosome in female predominance and in the deregulation of the immune system that characterizes autoimmunity. We have now evaluated X monosomy rates in SLE using fluorescence in situ hybridization (FISH) on peripheral mononuclear white blood cells (PBMCs) from female patients compared to healthy age-matched controls. In addition, because of a previous finding of microchimerism as a pathogenetic cause of a number of autoimmune diseases, we investigated the presence of cells carrying the Y chromosome. We did not identify an increased X monosomy in women with SLE compared to controls (P = 0.3960, SLE vs. HCs, Student's t-test), thus suggesting that a different mechanism of immune deregulation might be predominant in the female population of patients with SLE.

Personality type of the glaucoma patient.

PURPOSE: To characterize the personality profile of glaucoma subjects. PATIENTS AND METHODS: One hundred eight subjects including 56 open-angle glaucoma (OAG) and 52 controls were given the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) test and all performed automated perimetry. Clinical and demographic information which could relate to personality type was collected. RESULTS: OAG subjects had significantly higher Hypochondriasis (Hs; P=0.0082), Hysteria (Hy; P=0.0056), and Health Concerns (HEA; P=0.0025) mean scores than the control group. OAG subjects also had a significantly greater frequency of clinically abnormal score for hysteria (P=0.0262), and health concerns (P=0.0018). Multivariate analysis of variance revealed that Hypochondriasis, Hysteria, and Health Concerns scores were related to number of systemic medications used and to diagnostic group. Other potential explanatory variables such as sex, ethnicity, number of medical problems, length of glaucoma diagnosis, occurrence of glaucoma surgery, intraocular pressure, and visual status (logMAR, visual field indices) were not related to these personality scores. CONCLUSIONS: Patients with a diagnosis of OAG had more abnormal MMPI-2 scores in areas that focus upon concerns of somatic complaints and poor health. The use of systemic medications, which may be a constant reminder of illness, is a factor that may contribute to higher MMPI-2 scores.

Portrayals of branded soft drinks in popular American movies: a content analysis.

BACKGROUND: This study examines the portrayals of soft drinks in popular American movies as a potential vehicle for global marketing and an indicator of covert product placement. METHODS: We conducted a content analysis of America's top-ten grossing films from 1991 through 2000 that included portrayals of beverages (95 movies total). Coding reliabilities were assessed with Cohen's kappa, and exceeded 0.80. If there was at least one instance of branding for a beverage, the film was considered having branded beverages. Fisher's exact test was used to determine if soft drink portrayals were related to audience rating or genre. Data on the amount of time soft drinks appeared onscreen was log transformed to satisfy the assumption of normality, and analyzed using a repeated measures ANOVA model. McNemar's test of agreement was used to test whether branded soft drinks are as likely to appear or to be actor-endorsed compared to other branded beverages. RESULTS: Rating was not associated with portrayals of branded soft drinks, but comedies were most likely to include a branded soft drink (p = 0.0136). Branded soft drinks appeared more commonly than other branded non-alcoholic beverages (p = 0.0001), branded beer (p = 0.0004), and other branded alcoholic beverages (p = 0.0006). Actors consumed branded soft drinks in five times the number of movies compared to their consumption of other branded non-alcoholic beverages (p = 0.0126). About half the revenue from the films with portrayals of branded soft drinks come from film sales outside the U.S. CONCLUSION: The frequent appearance of branded soft drinks provides indirect evidence that product placement is a common practice for American-produced films shown in the U.S. and other countries.

Frequency of monosomy X in women with primary biliary cirrhosis.

The mechanisms that cause the female predominance of primary biliary cirrhosis (PBC) are uncertain, but the X chromosome includes genes involved in immunological tolerance. We assessed the rate of X monosomy in peripheral white blood cells from 100 women with PBC, 50 with chronic hepatitis C, and 50 healthy controls, by fluorescence in-situ hybridisation. Frequency of X monosomy increased with age in all groups, but was significantly higher in women with PBC than in controls (p<0.0001); age-adjusted back-transformed mean frequencies were 0.050 (95% CI 0.046-0.055) in women with PBC, 0.032 (0.028-0.036) in those with chronic hepatitis C, and 0.028 (0.025-0.032) in controls. We suggest that haploinsufficiency for specific X-linked genes leads to female susceptibility to PBC.

A multidisciplinary community hospital program for early and rapid resuscitation of shock in nontrauma patients.

OBJECTIVE: To determine the effect of a community hospital-wide program enabling nurses and prehospital personnel to mobilize institutional resources for the treatment of patients with nontraumatic shock. DESIGN: Historically controlled single-center study. SETTING: A 180-bed community hospital. PATIENTS: Patients in shock who were candidates for aggressive therapy. INTERVENTIONS: From January 1998 to May 31, 2000, patients in shock received standard therapy (control group). During the month of June 2000, intensive education of all health-care providers (ie, prehospital personnel, nurses, and physicians) took place. From July 1, 2000, through June 30, 2001, patients in shock (protocol group) were managed with a hospital-wide shock program. The program included early recognition of shock and the initiation of therapy by nonphysicians. Frontline personnel mobilized a shock team, which used goal-directed resuscitation protocols, early intensivist involvement, and rapid transfer to the ICU where protocols specific to shock etiology were implemented. MEASUREMENTS AND MAIN RESULTS: Eighty-six and 103 patients, respectively, were enrolled in the control and protocol groups. Baseline characteristics were similar. The protocol group had significant reductions in the median times to interventions, as follows: intensivist arrival, 2:00 h to 50 min (p < 0.002); ICU/operating room admission, 2 h 47 min to 1 h 30 min (p < 0.002); 2 L fluid infused, 3 h 52 min to 1 h 45 min (p < 0.0001); and pulmonary artery catheter placement, 3 h 50 min to 2 h 10 min (p 0.02). Good outcomes (ie, discharged to home or to a rehabilitation center) were more likely in the protocol group than in the control group (p = 0.02). The hospital mortality rate was 40.7% in the control group and 28.2% in the protocol group (p = 0.035). CONCLUSION: Similar to current practice in patients who have experienced trauma or cardiac arrest, the empowerment of nonphysician providers to mobilize hospital resources for the care of patients with shock is effective. A community hospital program incorporating the education of providers, the activation of a coordinated team response, and early goal-directed therapy expedited appropriate treatment and was temporally associated with improved outcomes. Randomized multicenter trials are needed to further assess the impact of the shock program on outcomes.

The long-term financial and clinical impact of an electronic health record on an academic ophthalmology practice.

Purpose. To examine financial and clinical work productivity outcomes associated with the use of the electronic health record (EHR). Methods. 191,360 billable clinical encounters were analyzed for 12 clinical providers over a 9-year study period during which an EHR was implemented. Main outcome measures were clinical revenues collected per provider and secondary outcomes were charge capture, patient visit coding levels, transcription costs, patient visit volume per provider, digital drawing, and digital imaging volume. Results. The difference in inflation adjusted net clinical revenue per provider per year did not change significantly in the period after EHR implementation (mean = $404,198; SD = $17,912) than before (mean = $411,420; SD = $39,366) (P = 0.746). Charge capture, the proportion of higher- and lower-level visit codes for new and established patients, and patient visits per provider remained stable. A total savings of $188,951 in transcription costs occurred over a 4-year time period post-EHR implementation. The rate of drawing the ophthalmic exam in the EHR was low (mean = 2.28%; SD = 0.05%) for all providers. Conclusions. This study did not show a clear financial gain after EHR implementation in an academic ophthalmology practice. Ophthalmologists do not rely on drawings to document the ophthalmic exam; instead, the ophthalmic exam becomes text-driven in a paperless world.

Titers of antibody to common pathogens: relation to food-based interventions in rural Kenyan schoolchildren.

BACKGROUND: Undernutrition is widely perceived to affect the development of an effective immune system. OBJECTIVE: We used a mini-analysis system to quantitate antibody titers and evaluate the sera of 200 Kenyan schoolchildren for antibodies to Helicobacter pylori [isotypes of immunoglobulins A (IgA), G (IgG), and M (IgM)], hepatitis A virus, rotavirus, tetanus toxoid (IgG), and a panel of recombinant malarial antigens (MSP1(19), MSP2, Ag512, MSP4, and MSP5). DESIGN: Children participated in a school-based feeding intervention with meat, milk, or nonanimal-source foods or in a nonintervention control group. Microvolumes (200 mL) of sera were analyzed at baseline and after 1 y. RESULTS: Nearly all children had elevated titers of antibody to H. pylori, hepatitis A virus, rotavirus, and malaria at the outset, despite a high prevalence of apparent biochemical micronutrient deficiencies and stunting, but many had titers of tetanus toxoid IgG antibodies below the protective concentration. Children with low hemoglobin had a greater proportion of elevated H. pylori IgM antibody titers at baseline, which suggests that current infection with H. pylori may be associated with anemia. Compared with the control subjects, only the group eating meat had a significant increase in H. pylori IgM antibodies during the intervention (P = 0.019). No other group comparisons with the control subjects were statistically significant. The additional finding that the sera of some children showed inadequate tetanus-protective antibodies, despite immunization, suggests that the vaccination program was suboptimal. CONCLUSIONS: A large battery of immune assays can be performed on microvolumes of sera. Furthermore, despite evidence of malnutrition, children do develop significant antibody-mediated responses to common pathogens.

Tenofovir treatment at 30 mg/kg/day can inhibit cortical bone mineralization in growing rhesus monkeys (Macaca mulatta).

The acyclic nucleoside phosphonate analog, 9-[2-(R)-(phosphonomethoxy)propyl]adenine (PMPA; Tenofovir: Gilead Sciences, Inc., Foster City, CA), has been shown to effectively inhibit simian immunodeficiency virus (SIV) replication in rhesus macaques by blocking reverse transcription. However, chronic long-term tenofovir treatment at 30 mg/kg/day, intended to reduce viral replication and illness, has been shown to result in bone deformities and spontaneous fractures in rhesus monkeys. Based on these findings, we studied the effects of tenofovir treatment and pathogenic SIV infection on cortical bone remodeling in rhesus monkeys. Tibiae from tenofovir-treated or untreated, SIV-infected or uninfected, rhesus macaques were evaluated for bone microdamage and remodeling. We found that tenofovir treatment had a significant effect on osteoid (unmineralized bone) seam width in tibial cross-sections. Regardless of SIV infection status, half of the tenofovir-treated animals had significantly increased osteoid seam widths in tibial cortical bone resulting in an osteomalacia-like condition. Pathogenic SIV infection significantly increased tibial resorption cavity density. and this increase was normalized by tenofovir treatment. These results suggest that tenofovir treatment at 30 mg/kg/day inhibits mineralization of newly formed bone. SIV infection results in increased tibial resorption cavity density, while tenofovir treatment tends to minimize this increase. Both defective mineralization of newly formed bone and increased resorption cavity density may result in greater bone fragility.

Adherence to glaucoma medication: the effect of interventions and association with personality type.

PURPOSE: To determine whether multiple interventions influence adherence to glaucoma medication and to study the relationship between personality type and adherence. METHODS: Eighty patients with glaucoma using prostaglandin monotherapy were prospectively randomized to an intervention or a nonintervention group. Over a 5-month period, the intervention group received monthly automated telephone calls reminding them to take their glaucoma medications. At month 3, the intervention group had an office visit with their physician and received education regarding their disease and treatment. The nonintervention group returned at the end of the study period. Adherence was directly measured with an electronic monitoring cap. Outcome measures were adherence rate and therapeutic coverage. Patients completed the Minnesota Multiphasic Personality Inventory-2. RESULTS: Thirty-eight patients were randomized to the intervention group and 42 were randomized to the nonintervention group. Mean adherence rate for the intervention group over 5 months was 76% and for the nonintervention group, 81%. For the intervention group, adherence did not change significantly after the midstudy visit (P=0.233). No difference existed between the intervention and nonintervention groups before (P=0.456) or after (P=0.134) the midstudy intervention visit. The Minnesota Multiphasic Personality Inventory-2 personality scales for depression (P=0.01) and hypochondriasis (P>0.0001) were significantly associated with poorer adherence. CONCLUSIONS: Monthly automated telephone reminders, a 1-time educational session, and increased contact with a physician did not improve adherence rate with glaucoma medications. Depression and hypochondriasis personality types were related to poor adherence and highlight the role of psychosocial factors in medication adherence.

Risk factors and comorbidities in primary biliary cirrhosis: a controlled interview-based study of 1032 patients.

Primary biliary cirrhosis (PBC) is an autoimmune disease of unknown etiology, often associated with other autoimmune conditions. Controlled studies have so far provided conflicting data on risk factors and comorbidity rates in PBC. We enrolled patients with PBC (n = 1032) from 23 tertiary referral centers for liver diseases in the United States and random-digit-dialed controls (n = 1041) matched for sex, age, race, and geographical location. Patients and controls were administered a modified version of the US National Health and Nutrition Examination Study (NHANES III) questionnaire by trained personnel to evaluate associations between PBC and social, demographic, personal and family medical histories, lifestyle, and reproductive factors and the rates of comorbidity in affected individuals. Data indicate that having a first-degree relative with PBC (adjusted odds ratio [AOR] 10.736; 95% confidence interval 4.227-27.268), history of urinary tract infections (AOR 1.511, 95% CI 1.192-1.915), past smoking (AOR 1.569, 95% CI 1.292-1.905), or use of hormone replacement therapies (AOR 1.548, 95% CI 1.273-1.882) were significantly associated with increased risk of PBC. The frequent use of nail polish slightly increased the risk of having PBC. Other autoimmune diseases were found in 32% of cases and 13% of controls (P<0.0001). In conclusion, environmental factors, possibly including infectious agents through urinary tract infections or chemicals contained in cigarette smoke, may induce PBC in genetically susceptible individuals. Exogenous estrogens may also contribute to explain the female predominance of the disease.

X chromosome monosomy: a common mechanism for autoimmune diseases.

The majority of human autoimmune diseases are characterized by female predominance. Although sex hormone influences have been suggested to explain this phenomenon, the mechanism remains unclear. In contrast to the role of hormones, it has been suggested, based on pilot data in primary biliary cirrhosis, that there is an elevation of monosomy X in autoimmune disease. Using peripheral white blood cells from women with systemic sclerosis (SSc), autoimmune thyroid disease (AITD), or healthy age-matched control women, we studied the presence of monosomy X rates using fluorescence in situ hybridization. We also performed dual-color fluorescence in situ hybridization analysis with a chromosome Y alpha-satellite probe to determine the presence of the Y chromosome in the monosomic cells. In subsets of patients and controls, we determined X monosomy rates in white blood cell subpopulations. The rates of monosomy X increased with age in all three populations. However, the rate of monosomy X was significantly higher in patients with SSc and AITD when compared with healthy women (6.2 +/- 0.3% and 4.3 +/- 0.3%, respectively, vs 2.9 +/- 0.2% in healthy women, p < 0.0001 in both comparisons). Importantly, X monosomy rate was more frequent in peripheral T and B lymphocytes than in the other blood cell populations, and there was no evidence for the presence of male fetal microchimerism. These data highlight the thesis that chromosome instability is common to women with SSc and AITD and that haploinsufficiency for X-linked genes may be a critical factor for the female predominance of autoimmune diseases.

Genetic polymorphisms influencing xenobiotic metabolism and transport in patients with primary biliary cirrhosis.

Epidemiological data suggest that environmental factors may trigger autoimmunity in genetically susceptible individuals. In primary biliary cirrhosis (PBC), it has been postulated that halogenated xenobiotics can modify self-molecules, facilitating the breakdown of tolerance to mitochondrial antigens. The transport and metabolism of xenobiotics is highly dependent on key genetic polymorphisms that alter enzymatic phenotype. We analyzed genomic DNA from 169 patients with PBC and 225 geographically and sex-matched healthy subjects for polymorphisms of genes coding for cytochromes P450 (CYPs) 2D6 (CYP2D6*4, CYP2D6*3, CYP2D6*5, and CYP2D6*6) and 2E1 (cl/c2), multidrug resistance 1 (MDR1 C3435T) P-glycoprotein, and pregnane X receptor (PXR C-25385T, C8055T, and A7635G). We compared the genotype frequencies in patients and controls and also correlated polymorphisms with PBC severity. The distributions of the studied genotypes did not significantly differ between patients and controls. However, when clinical characteristics of patients with PBC were compared according to genotype, the CYP2E1 c2 allele was associated with signs of more severe disease. In conclusion, genetic polymorphisms of CYP 2D6 and 2E1, PXR, and MDR1 do not appear to play a role in the onset of PBC.

Cardiac output monitoring during off-pump coronary artery bypass grafting.

OBJECTIVE: To evaluate and compare monitors of cardiac output during repositioning and stabilization of the heart for off-pump coronary artery bypass (OPCAB) surgery. DESIGN: Prospective, observational, clinical study. SETTING: University teaching hospital. PARTICIPANTS: Consecutive patients scheduled to undergo elective OPCAB (n = 19). INTERVENTIONS: Monitoring, induction, and anesthesia followed a routine protocol for coronary artery bypass patients. This included the use of transesophageal echocardiography (TEE) and pulmonary artery catheter placement. MEASUREMENTS AND MAIN RESULTS: After positioning and stabilization for OPCAB surgery, the changes in descending aortic flow velocity (VTI) times heart rate (HR) and the mixed venous oxygen saturation (SvO(2)) could be used to predict the changes in thermodilution cardiac output (TDCO) using the following model: deltaTDCO((calc))=-13.15+0.35(deltaVTI*HR)+0.61(deltaSvO(2)) where Delta indicates the percentage change from baseline values. The changes in mean arterial pressure, mean pulmonary artery pressure, and continuous cardiac output did not correlate with the changes in TDCO. CONCLUSION: The use of the VTI*HR, as determined by TEE, in addition to the SvO(2) can strengthen clinical decision making during repositioning and stabilization of the heart during OPCAB. Changes in the VTI*HR and SvO(2) can be used as surrogate markers for changes in CO during OPCAB surgery.

Alanine racemase free energy profiles from global analyses of progress curves.

Free energy profiles for alanine racemase from Bacillus stearothermophilus have been determined at pH 6.9 and 8.9 from global analysis of racemization progress curves. This required a careful statistical design due to the problems in finding the global minimum in mean square for a system with eight adjustable parameters (i.e., the eight rate constants that describe the stepwise chemical mechanism). The free energy profiles obtained through these procedures are supported by independent experimental evidence: (1). steady-state kinetic constants, (2). solvent viscosity dependence, (3). spectral analysis of reaction intermediates, (4). equilibrium overshoots for progress curves measured in D(2)O, and (5). the magnitudes of calculated intrinsic kinetic isotope effects. The free energy profiles for the enzyme are compared to those of the uncatalyzed and the PLP catalyzed reactions. At pH 6.9, PLP lowers the free energy of activation for deprotonation by 8.4 kcal/mol, while the inclusion of apoenzyme along with PLP additionally lowers it by 11 kcal/mol.

Suppression of leukotriene B4 generation by ex-vivo neutrophils isolated from asthma patients on dietary supplementation with gammalinolenic acid-containing borage oil: possible implication in asthma.

Dietary gammalinolenic acid (GLA), a potent inhibitor of 5-lipoxygenase (5-LOX) and suppressor of leukotriene B4 (LTB4), can attenuate the clinical course of rheumatoid arthritics, with negligible side effects. Since Zileuton, also an inhibitor of 5-LOX, attenuates asthma but with an undesirable side effect, we investigated whether dietary GLA would suppress biosynthesis of PMN-LTB4 isolated from asthma patients and attenuate asthma. Twenty-four mild-moderate asthma patients (16-75 years) were randomized to receive either 2.0 g daily GLA (borage oil) or corn oil (placebo) for 12 months. Blood drawn at 3 months intervals was used to prepare sera for fatty acid analysis, PMNs for determining phospholipid fatty acids and for LTB4 generation. Patients were monitored by daily asthma scores, pulmonary function, and exhaled NO. Ingestion of daily GLA (i) increased DGLA (GLA metabolite) in PMN-phospholipids; (ii) increased generation of PMN-15-HETrE (5-LOX metabolite of DGLA). Increased PMN-DGLA/15-HETrE paralleled the decreased PMN generation of proinflammatory LTB4. However, the suppression of PMN-LTB4 did not reveal statistically significant suppression of the asthma scores evaluated. Nonetheless, the study demonstrated dietary fatty acid modulation of endogenous inflammatory mediators without side effects and thus warrant further explorations into the roles of GLA at higher doses, leukotrienes and asthma.

Serial analysis of antimitochondrial antibody in patients with primary biliary cirrhosis.

Antimitochondrial antibodies (AMAs) are the classic serologic marker in primary biliary cirrhosis (PBC). However, there have been only limited attempts to study changes in titer or isotype analysis of such AMAs in patients followed for long periods of time. We took advantage of stored sera from well-characterized patients with PBC followed for a period of 7-28 years (mean duration of 13.5 years). Immunoblot and enzyme-linked immunosorbant assays were performed against PDC-E2, BCOADC-E2 and OGDC-E2 as well as isotype analysis of antigen-specific IgG, IgA and IgM antibodies against each of these mitochondrial autoantigens. Sera were analyzed for total IgG, IgA and IgM by radial immunodiffusion. The sera titer of AMAs was significantly higher in younger patients with PBC. Indeed, age of onset of clinical PBC was a significant predictor for the highest values of sera AMAs. In contrast, the AMA titer did not significantly change over time in this prolonged longitudinal study. The total sera levels of the individual immunoglobulins did not show a time-dependent change, when based on age of onset of the disease. Higher titers of AMAs were noted in the younger patients. Furthermore, despite this long follow-up, there was no evidence for a significant change in AMA levels; also, levels were not influenced by drug therapy used during the period of observation.

Highly tissue substructure-specific effects of human papilloma virus in mucosa of HIV-infected patients revealed by laser-dissection microscopy-assisted gene expression profiling.

Human papilloma virus (HPV) causes focal infections of epithelial layers in skin and mucosa. HIV-infected patients on highly active antiretroviral therapy (HAART) appear to be at increased risk of developing HPV-induced oral warts. To identify the mechanisms that allow long-term infection of oral epithelial cells in these patients, we used a combination of laser-dissection microscopy (LDM) and highly sensitive and quantitative, non-biased, two-step multiplex real-time RT-PCR to study pathogen-induced alterations of specific tissue subcompartments. Expression of 166 genes was compared in three distinct epithelial and subepithelial compartments isolated from biopsies of normal mucosa from HIV-infected and non-infected patients and of HPV32-induced oral warts from HIV-infected patients. In contrast to the underlying HIV infection and/or HAART, which did not significantly elaborate tissue substructure-specific effects, changes in oral warts were strongly tissue substructure-specific. HPV 32 seems to establish infection by selectively enhancing epithelial cell growth and differentiation in the stratum spinosum and to evade the immune system by actively suppressing inflammatory responses in adjacent underlying tissues. With this highly sensitive and quantitative method tissue-specific expression of hundreds of genes can be studied simultaneously in a few cells. Because of its large dynamic measurement range it could also become a method of choice to confirm and better quantify results obtained by microarray analysis.