Novel atherogenic pathways from the differential transcriptome analysis of diabetic epicardial adipose tissue.

BACKGROUND AND AIM: To evaluate the epicardial adipose tissue (EAT) transcriptome in comparison to subcutaneous fat (SAT) in coronary artery disease (CAD) and type 2 diabetes (T2DM). METHODS AND RESULTS: SAT and EAT samples were obtained from subjects with T2DM and CAD (n = 5) and those without CAD with or without T2DM (=3) undergoing elective cardiac surgery. RNA-sequencing analysis was performed in both EAT and SAT. Gene enrichment analysis was conducted to identify pathways affected by the differentially expressed genes. Changes of top genes were verified by quantitative RT-PCR (qRT-PCR), western blot, and immunofluorescence. A total of 592 genes were differentially expressed in diabetic EAT, whereas there was no obvious changes in SAT transcriptome between diabetics and non-diabetics. Diabetic EAT was mainly enriched in inflammatory genes, such as Colony Stimulating Factor 3 (CSF3), Interleukin-1b (IL-1b), IL-6. KEGG pathway analysis confirmed that upregulated genes were involved in inflammatory pathways, such as Tumor Necrosis Factor (TNF), Nuclear Factor-κB (NF-κB) and advanced glycation end-products-receptor advanced glycation end products (AGE-RAGE). The overexpression of inflammatory genes in diabetic EAT was largely correlated with upregulated transcription factors such as NF-κB and FOS. CONCLUSIONS: Diabetic EAT transcriptome is significantly different when compared to diabetic SAT and highly enriched with genes involved in innate immune response and endothelium, like Pentraxin3 (PTX3) and Endothelial lipase G (LIPG). EAT inflammatory genes expression could be induced by upregulated transcription factors, mainly NF-kB and FOSL, primarily activated by the overexpressed AGE-RAGE signaling. This suggests a unique and novel atherogenic pathway in diabetes.

Alcohol consumption in late-life--the first Brazilian National Alcohol Survey (BNAS).

To investigate the alcohol consumption in later life in Brazil and its association with socio-demographic characteristics. This study was a cross-sectional analysis of nationally representative survey data. A multistage cluster sampling procedure was used to select 3007 individuals of 14 years of age and older from the Brazilian household population. In this study we analyzed data from all 400 participants who were over 60 years old. Alcohol Abuse and Dependence Syndrome was established according to DSM-IV and Risky Drinking was defined in two ways: heavy drinkers (>7 drinks/week) and as binge drinkers (>3 drinks/one occasion). Twelve percent of participants reported heavy drinking behavior while 10.4% and 2.9% were binge drinkers and alcohol dependent respectively. In the adjusted logistic regression only gender was associated with heavy drinking behavior. Males, the youngest and the wealthiest were more likely to report binge drinking behaviors. In conclusion, alcohol related-problems are common but under recognized among older adults. Health professionals should be aware that common definitions of alcohol abuse and dependence may not apply as readily to older people, who have had biological changes for alcohol tolerance and its effects on the Central Nervous System.

Utility of 2-D and 3-D virtual microscopy in cervical cytology education and testing.

OBJECTIVE: To evaluate the effectiveness of 3-D vs. 2-D virtual microscopy as adjuncts to education and assessment in cervical cytology. STUDY DESIGN: Five cervical cytology slides were acquired in 2-D; then the identical area of the slide was acquired in 3-D, resulting in 2 sets of virtual slides for comparison with the original glass slide. Seventy-nine paid volunteer cytologists and cytotechnology students participated. Approximately half were sent the 2-D set of slides via the Web, and the others a 3-D set of slides on a DVD. Evaluators examined the virtual slides and committed to an interpretation. After receipt of the original glass slides, a second interpretation was made, if different from the virtual slide interpretation. RESULTS: Diagnostic accuracy using virtual cytology slides was similar to that for glass slides (94% vs. 96%). There was no difference in diagnostic accuracy between 2-D and 3-D slides (p = 0.28); however, the ability to focus 3-D slides in the z-axis was strongly endorsed by the participants because of the uncertainty and frustration of having some cells out of focus on 2-D virtual slides. CONCLUSION: There was consensus that virtual cervical cytology slides would be a useful augmentation to education and testing.

Variability in cytologic-histologic correlation practices and implications for patient safety.

CONTEXT: The Clinical Laboratory Improvement Amendments of 1988 require that laboratories perform cytologic-histologic correlation, although the optimal methods and the value of performing correlation have not been determined. OBJECTIVE: To determine the similarities and differences in how laboratories perform cytologic-histologic correlation. DESIGN: One hundred sixty-two American laboratories were sent a letter requesting copies of the materials they used in the cytologic-histologic correlation process. The returned materials were classified into the categories of forms, logs, and tally sheets. A checklist (derived from the College of American Pathologists Laboratory Accreditation Cytopathology Checklist) was developed to classify the "minimum expected" (15) and "additional" data points that laboratories collected when they performed a correlation. PARTICIPANTS: American pathology laboratories. MAIN OUTCOME MEASURES: Measures were percentage of laboratories that recorded minimum expected and additional data points and the frequency with which specific minimum expected data points were recorded. RESULTS: The response frequency was 32.1%, and a total of 84 cytologic-histologic correlation materials were obtained. The only minimum expected variables recorded on forms or logs by more than 50% of laboratories were cytology case number, sign-out cytology diagnosis, surgical pathology case number, and sign-out surgical pathology diagnosis. Nine (17.3%) laboratories did not record data on forms, logs, or tally sheets. The mean number of minimum expected and additional variables recorded on forms was 6.5 and 8.7, respectively. CONCLUSIONS: Laboratories record data from the cytologic-histologic correlation process in a number of ways, indicating the lack of standardization of the data collection process.

Improving Pap test turnaround time using external benchmark data and engineering process improvement tools.

Turnaround time for Papanicolaou (Pap) tests became an important service quality issue at our institution. We studied Pap test turnaround time using engineering process improvement tools and benchmarked turnaround time against data published as a College of American Pathologists Q-Probes study. An IDEF3 process map revealed the complexity of the Pap test process and the opportunities for process improvement. We used these data and the action-research method to initiate changes in cytopathology laboratory operations with the goal of reducing turnaround time. Before intervention, mean Pap test turnaround time was highly variable; during a 6-month period, monthly means ranged from 2.5 to 10.8 days. A cycle time study conducted over a 2-week period validated these data. After system improvements were implemented, the monthly mean turnaround time decreased and became more consistent, with 11 of 12 months having a mean turnaround time of 3 days or less (range, 1.5-3.9 days). Our study illustrates the value of publishing Q-Probes data for use as external benchmarks and the benefits of using tools from other disciplines to improve laboratory processes.

GABA(B) receptors play a role in the development of tolerance to ethanol in mice.

RATIONALE: There is evidence that drugs that improve or impair learning can facilitate or block ethanol tolerance, respectively. Since GABA(B) receptors have been shown to be involved in processes related to learning, it is possible that this system could play a role in the development of rapid tolerance to ethanol. OBJECTIVES: The aim of this study was to verify the influence of one GABA(B) agonist and two GABA(B) antagonists on tolerance to the effect of ethanol on motor coordination. METHODS: Male Swiss mice were trained on a continuously accelerating rota-rod device. Animals were pretreated with the GABA(B) agonist (-)-baclofen (3, 5, or 7 mg kg(-1)) or saline, 30 min before ethanol (1.75 g kg(-1)), and were tested 5, 10, and 15 min later on the rota-rod. In another set of experiments, mice were pretreated with the GABA(B) antagonists CGP36742 (1, 3, 10, or 30 mg kg(-1)) or CGP56433 (0.1, 0.3, 1.0, or 3.0 mg kg(-1)), or saline, 30 min before the test under ethanol. Rapid tolerance was evaluated 24 h after the first ethanol injection, by injecting all animals with ethanol and retesting them on the rota-rod. RESULTS: The results showed that (-)-baclofen (5 mg kg(-1)) significantly (ANOVA + Tukey's test) blocked rapid tolerance, whereas CGP36742 (3 and 10 mg kg(-1)) and CGP56433 (0.3, 1, and 3 mg kg(-1)) facilitated rapid tolerance in a dose-dependent way. The blockade of rapid tolerance by (-)-baclofen was antagonized by previous administration of CGP36742 or CGP56433. CONCLUSIONS: The current results suggest that rapid tolerance to ethanol is subjected to inhibition by a GABAergic GABA(B) receptor-mediated system in the mouse.

[Palatine tonsil lymphoma in ENT department in Belchatów for the of 1987-1998]. / Chloniaki zlosliwe migdalka podniebiennego w materiale Oddzialu Laryngologicznego Szpitala Rejonowego w Belchatowie w latach 1987-1998 r.

The authors present 10 cases of malignant lymphomas of the head and neck hospitalized in ENT Department of Regional Hospital in Belchatów. The two cases of tonsil lymphomas were described in detail calling attention to diagnostic difficulties. The false result of aspiration biopsy caused prolongation of diagnostic procedure.

Effect of cervical disease history on outcomes of women who have a pap diagnosis of atypical glandular cells of undetermined significance.

OBJECTIVE: To determine the relationship between history of cervical dysplasia or carcinoma and the development of cervical dysplasia or adenocarcinoma in women who have a diagnosis of atypical glandular cells of undetermined significance (AGUS), favor endocervical origin, or AGUS, not otherwise specified. METHODS: A 6-year retrospective review of the pathology files was performed for 93 women who were diagnosed in 1992 with AGUS, favor endocervical origin, or AGUS, not otherwise specified. Data collected included previous history of cervical disease, follow-up diagnoses, time interval between follow-ups, and procedures performed. RESULTS: Of women with follow-up who had or did not have a previous history of cervical dysplasia, 32.0 and 12.0%, respectively, developed a squamous dysplasia or adenocarcinoma in situ. This difference was statistically significant (P<0.05). Of the women who had or did not have a previous history of cervical dysplasia and had Pap smear follow-up, only 4.2 and 4.3%, respectively, had a false-negative diagnosis on the most immediate subsequent smear. CONCLUSIONS: Women who have AGUS, favor endocervical origin, or AGUS, not otherwise specified, and no history of cervical dysplasia have a significantly lower risk of developing or having cervical dysplasia than women who have the same diagnosis and a history of cervical dysplasia. This may warrant different treatment protocols for these two groups. For the women with AGUS and no previous history of cervical dysplasia, a repeat Pap smear, rather than colposcopy with curettage, may be warranted.

Long-term outcome and relative risk in women with atypical squamous cells of undetermined significance.

Few studies have compared long-term follow-up and risk for invasive cancer in women with atypical squamous cells of undetermined significance (ASCUS). We conducted a 6-year review of pathology files for 651 women in whom ASCUS had been diagnosed in 1992. Data collected included patient demographics, follow-up diagnoses, time between follow-up examinations, and procedures performed. At follow-up, high-grade squamous intraepithelial lesions (HSIL) had developed in 9.0% of the women, and invasive cancer in none. Previous cervical history did not affect risk for an HSIL. Although the average time to first follow-up was 6.18 months, in 20.9% of the women the diagnosis of HSIL was not established until after 2.0 years. For individual pathologists, the percentage of HSILs ranged from 0% to 18.8%. Thus women with ASCUS who are followed up regularly are at low risk for development of invasive cancer.

Cost effectiveness of rescreening cervicovaginal smears.

Although cytology laboratories are mandated to rescreen at least 10% of cervicovaginal smears, there is no uniform national rescreening practice. Follow-up data for 16,188 rescreened cervicovaginal smears were studied and decision analysis was performed to determine an optimal rescreening strategy. High-grade dysplasia was detected in 0.40% of women with a history of cervical disease and in 0.04% without a history of cervical disease. Compared with 0% rescreening of smears, with 15% rescreening the cost to gain a year of discounted life expectancy was $386,890 for women without a history of cervical disease, and $2,980 for women with a history of cervical disease. We conclude that rescreening only smears from women with a history of cervical disease could save US laboratories more than $11.2 million annually without seriously compromising care.

The cost-effectiveness of the cytology laboratory and new cytology technologies in cervical cancer prevention.

The effect of changes in cytology laboratory costs, including the costs of new technologies, on the cost-effectiveness of cervical cancer prevention has not been studied. Using University of Iowa laboratory detection rates and costs, a decision model determined the cost-effectiveness of the laboratory with and without new technologies. Compared with not performing a cervicovaginal smear, the cost to increase the discounted life expectancy per patient by 1 year was $2,805 for the laboratory component alone and $19,655 for the entire cervical cancer prevention strategy. In moderate- to high-risk women, cervical cancer screening was cost-effective even at high cytology laboratory costs (eg, $75 per smear). New technologies were cost-effective only if they resulted in a substantial increase in the detection of high-grade squamous intraepithelial lesions (eg, an additional 236 high-grade squamous intraepithelial lesions per 10,000 women). New technologies have not demonstrated these increased detection rates.

Incidence of hypercalcaemia in haemodialysis patients.

Secondary hyperparathyroidism is still one of the main problems affecting the dialysis population. The use of aluminium hydroxide as a phosphate binder is limited by significant long-term toxicity. Therefore, actually, most patients on haemodialysis receive treatment with calcium containing phosphate binders to avoid hyperphosphataemia.

Telecytology: diagnostic accuracy in cervical-vaginal smears.

Although cervical-vaginal telecytology is a promising tool, diagnostic accuracy has not been extensively evaluated. The authors examined the accuracy of five cytotechnologists who retrospectively reviewed 50 cervical-vaginal smears using the video monitor, and 2 months later, using the light microscope. Accuracy was expressed in terms of crude agreement with the original diagnosis and number of false positives (FPs) and false negatives (FNs). With a greater than one step difference as discrepant, the group crude agreement using the video monitor and the light microscope was 85.6% and 95.6%, respectively. The group number of FNs and FPs for the light microscope was 8 and 7, respectively, and for the video monitor was 34 and 7, respectively. There was a wide range of individual performance. We conclude that accuracy of telecytology is high, but less than that of light microscopy. The major reason for lower telecytologic accuracy was undercalling dysplasia.

Cytomorphologic characteristics of thermal injury related to endocervical brushing following loop electrosurgical excision procedure (LEEP).

Loop electrosurgical excision procedure (LEEP) is gaining popularity in the United States as an alternative to other ablative and cone methods of treatment for preneoplastic conditions of the uterine cervix. The major advantage is that it is an outpatient procedure using local anesthesia. Post-LEEP evaluation of the endocervical canal can be accomplished by either endocervical curettage or cytobrush. We reviewed the immediate post-LEEP cytobrushes from 33 patients. Artifact related to the procedure was seen in all cases and included: 1) elongated endocervical cells with cytoplasmic "tails" and streaming nuclei ("taffy pulled"); 2) loose cellular aggregates with coalesced cytoplasm and "hockey stick" nuclei; 3) sheets of cells with frothy cytoplasm and shrunken thumbprinted nuclei; 4) nuclear enlargement; and 5) smudgy nuclear chromatin. The background ranged from bloody coagulum to watery proteinaceous material. Original diagnoses were negative in 25 cases. Eleven (44%) were considered abnormal on review. Of these, four were interpreted as having squamous intraepithelial lesions (SIL), one was atypical squamous cells of undetermined significance (ASCUS), and six had atypical glandular cells of undetermined significance (AGUS). It was not always possible to grade the degree of dysplasia. There was review agreement with 14 negatives, one SIL, one ASCUS, and one unsatisfactory specimen. Three AGUS were upgraded to SIL, and one unsatisfactory specimen was considered atypical on review. In conclusion it is possible to separate artifact from actual pathology on post-LEEP cytobrush specimens. Recognition of the characteristic changes seen on cytologic specimens obtained immediately following diathermy loop treatment will allow accurate identification of true abnormalities.

Retrovirus-induced lymphoproliferative disease in mice undergoing graft-versus-host reaction.

The effect of graft-versus-host reaction on the course of concommitant retrovirus-induced lymphoproliferative disease was investigated. The graft-versus-host reaction was elicited by a single i.v. injection of 1.2 x 10(8) parental spleen cells into adult F1 mice. Lymphoproliferative disease was induced by a single transfusion of 0.2 ml of whole blood from donors with fully developed disease, induced by infection with retrovirus LP-BM5 MuLV. Graft-versus-host reaction and the lymphoproliferative disease each separately produced similar syndrome consisting of splenomegaly, lymphadenopathy, leukopenia, neutrophilia, reduced in vitro proliferation of spleen cells and suppression of in vivo immune responsiveness. The above symptoms were usually less pronounced during graft-versus-host reaction. Ongoing graft-versus-host reaction neither aggravated nor accelerated the course of the virus-induced lymphoproliferative disease in genetically susceptible F1 hybrids. Likewise, an ongoing graft-versus-host reaction in genetically resistant F1 hybrids did not alter their susceptibility to the retrovirus infection. The apparent lack of the effect of graft-versus-host reaction -dependent immunosuppression on the severity and the course of the concommitant retrovirus-induced lymphoproliferative disease suggests pathogenic differences between the murine syndrome and human AIDS for which the murine disease is considered by some to be an animal model.

Retrovirus-induced lymphoproliferative disease in mice: role of humoral immunity in perinatally exposed mice.

The mice born to female mice infected with LP-BM5 MuLV, the etiologic agent for lymphoproliferative disease and nursed for 4-6 weeks by them were less susceptible upon reinfection by i.v. transfusion of blood or plasma from infected donors with fully developed disease. Sera of 7 week or older perinatally exposed mice were capable of a complete in vitro neutralization of virus in plasma or blood from mice with fully developed disease. In contrast, sera from 3-week old perinatally exposed mice were ineffective. The neutralizing ability of the sera was drastically reduced or abrogated after their absorption with anti-mouse IgM. These observations are consistent with the notion that perinatally exposure results ina moderate form of the disease of the offspring. This perinatal infection is followed by a production of neutralizing antibodies of predominantly the IgM class that significantly alters the course of the lymphoproliferative disease and, in some instances, even prevents its development.

Pathology and probability. Likelihood ratios and receiver operating characteristic curves in the interpretation of bronchial brush specimens.

Diagnoses in pathology often are qualitative, such as atypical or suspicious, and consequently are thought to have limited clinical value. To investigate the utility of a qualitative diagnostic system, seven pathologists retrospectively evaluated 100 bronchial brush specimens using the following categories: definitely benign, probably benign, possibly malignant, probably malignant, and definitely malignant. The likelihood ratio (LR) and receiver operating characteristic (ROC) curve, two statistical probabilistic measurements, were used to calculate diagnostic accuracy among individuals and groups. The results show: (1) the LR for individual diagnostic categories varied among observers, resulting in different clinically malignant probabilities; (2) observer experience did not appear to play a role in overall diagnostic accuracy, except in the diagnosis of small cell carcinoma; (3) observers operate at higher levels of diagnostic accuracy with, rather than without, clinical history. The authors conclude that qualitative diagnoses contain important information and can be interpreted effectively with LR and ROC.

Laboratory use of the diagnosis "reactive/reparative" in gynecologic smears: impact of CLIA '88.

The Clinical Laboratory Improvement Amendments of 1988 (CLIA '88) contain regulation 493.1257(c)(1), which states: "[A]ll gynecologic smears interpreted to be showing reactive or reparative changes are confirmed by a technical supervisor in cytology." We surveyed 500 cytology laboratories to assess the utilization of the diagnosis "reactive/reparative" (R/R) for gynecologic cytology examinations. One hundred and seventy participants (34%) returned an acceptable questionnaire that described their use of this terminology, the technical supervisor's (pathologist's) role in the evaluation of these cases, and the volume of cases with this diagnosis. Prior to CLIA '88, 86 (51%) laboratories did not forward to a pathologist gynecologic smears reported as showing R/R changes, and this decreased to 20 (12%) after implementation of CLIA '88. Thirty-six (21%) laboratories changed their criteria for reporting R/R changes after CLIA '88. No correlation was noted between the demographics of the laboratory (e.g., size, type of practice affiliation, accreditation) and familiarity with this regulation. Based on the case volumes provided in the survey results, the estimated cost to the nation of complying with this single regulation is between $4.5 million and $40 million. The interpretation of and financial impact of compliance with this one selected regulation of CLIA '88 is reported.