bluuebunny/crossref_metadata_2025 huggingface dataset:
The hair follicle barrier to involvement by malignant melanoma
Authors: Pozdnyakova Olga , Grossman Joseph , Barbagallo Belinda , Lyle Stephen
Published: January 2009

BACKGROUND:

Melanoma characteristically grows within the epidermis along the dermal‐epidermal junction, sometimes extending outward up to several centimeters beyond the foci of invasive tumors. Although follicular involvement by malignant melanoma is widely recognized, to the authors' knowledge no previously published data address this phenomenon.

METHODS:

To examine the growth characteristics of in situ melanomas in relation to the hair follicle microanatomy, the authors analyzed 100 cases of primary cutaneous melanomas (61 in situ and 39 invasive melanomas with significant in situ components) obtained from pathology clinical archives.

RESULTS:

Eighty‐two (82%) cases of melanoma in situ demonstrated tumor cells within ≥1 hair follicles. Of those, 57 (69.5%) cases demonstrated the tumor cells only within the infundibulum. Extension of the tumor cells down to the isthmus was observed in 24 cases (29.3%). In only 1 exceptional case (1%) were tumor cells detected beneath the level of the hair follicle bulge.

CONCLUSIONS:

The authors postulate that a physiologic barrier restricts the intraepithelial spread of melanoma tumor cells at or beyond the level of the stem cell niche in the hair follicle bulge. Although the nature of this barrier remains to be elucidated, the distinct biologic characteristics of the hair follicle bulge may provide clues to understanding this phenomenon. Cancer 2009. © 2009 American Cancer Society.
High prevalence of BRAF mutation in a Brazilian cohort of patients with sporadic papillary thyroid carcinomas
Authors: Oler Gisele , Cerutti Janete M.
Published: January 2009

BACKGROUND:

Although several studies undoubtedly demonstrated that BRAF mutation is an important genetic event in the pathogenesis of papillary thyroid carcinoma (PTC), its prognostic significance and correlation with less differentiated states remains unclear. It has been suggested that the discrepancy may be at least partially due to the insufficient number of cases analyzed, epidemiologic factors, and a combination of different variants of PTC included in these studies.

METHODS:

In this context, the prevalence of the BRAF mutation in a Brazilian cohort of PTCs (n = 120) was first assessed and correlated with clinicopathologic features. The BRAF exon 15 mutation was evaluated by direct sequencing. Furthermore, using quantitative polymerase chain reaction, the issue of whether the expression level of the iodide‐metabolizing genes (NIS and TSHR) was correlated with BRAF mutational status was investigated.

RESULTS:

A high prevalence of the BRAF mutation was found in PTC cases (48%). The BRAF mutation was found to be significantly associated with the classic variant of PTC (66%; P < .0001), although it was found in the follicular variant as well (21%). Subtype stratification demonstrated that the BRAF V600E mutation was associated with tumor size, extrathyroid invasion, the presence of lymph node metastasis and risk of disease recurrence, and mortality in patients with the classic variant of PTC. Moreover, the expression levels of NIS and TSHR were remarkably lower in PTCs harboring the BRAF V600E mutation.

CONCLUSIONS:

These findings provide further evidence that BRAF might be associated with a more aggressive phenotype and less differentiated state due to decreased expression of iodide‐metabolizing genes. The search for a BRAF mutation in the current study population appears to be valuable for predicting prognosis and guiding management in patients with PTC. Cancer 2009. © 2009 American Cancer Society.
Factors associated with local and distant recurrence and survival in patients with resected nonsmall cell lung cancer
Authors: Varlotto John M. , Recht Abram , Flickinger John C. , Medford‐Davis Laura N. , Dyer Ann M. , DeCamp Malcolm M.
Published: January 2009

BACKGROUND:

This study assessed the impact of surgical, histopathologic and patient‐related factors on the risks of local and distant recurrence and overall survival for patients with stages I through IIIA nonsmall cell lung carcinoma (NSCLC) undergoing definitive resection with or without adjuvant chemotherapy.

METHODS:

This study included 373 consecutive patients treated between 2000 and 2005 who did not receive adjuvant or neoadjuvant radiotherapy, had at least 3 months of follow‐up, and did not have a history of other cancers within 5 years of the diagnosis of their NSCLC. Of these, 52% had pathologic stage IA disease, 30% had stage IB, 5% had stage IIA, 8% had stage IIB and 5% had stage III disease. Forty‐four patients received chemotherapy.

RESULTS:

The median follow‐up was 33 months. Local failure rates at 2 years, 3 years, and 5 years were 16%, 22%, and 32%, respectively; distant recurrence rates were 13%, 15%, and 21%, respectively. Multivariable analysis revealed that local recurrence was significantly associated with the presence of lymphatic or vascular invasion (LVI), the use of chemotherapy, and having diabetes; distant recurrence was significantly higher in patients with nonsquamous cell histology, those undergoing pneumonectomy, and those with more advanced TNM stage. Survival was significantly associated with age, history of myocardial infarction, performance of a pneumonectomy, histology, LVI, and the number of positive N1 lymph nodes.

CONCLUSIONS:

Local recurrence was the predominant type of failure in this series. Patient with diabetes or LVI may benefit from close surveillance and aggressive therapy of asymptomatic local recurrences, especially when chemotherapy is given in addition to surgery. Cancer 2009. © 2009 American Cancer Society.
Understanding bladder cancer death
Authors: Morris David S. , Weizer Alon Z. , Ye Zaojun , Dunn Rodney L. , Montie James E. , Hollenbeck Brent K.
Published: January 2009

BACKGROUND:

To the authors' knowledge, the extent to which death from bladder cancer is attributable to tumor biology or physician practice patterns is unknown. For this reason, the relative importance of broadening indications for aggressive therapy has unclear implications.

METHODS:

Patients whose deaths were caused directly by bladder cancer were identified using institutional (n = 126 patients) and administrative (n = 6326 patients) data sources. By using implicit review (clinical data, 2001‐2005) and explicit algorithms (Surveillance, Epidemiology, and End Results [SEER]‐Medicare, 1992‐2002), the authors estimated the proportion of potentially avoidable deaths from bladder cancer.

RESULTS:

After an implicit review of clinical data, 40 of 126 deaths (31.7%) were classified as potentially avoidable. Compared with those patients who were deemed unsalvageable, these patients generally presented with nonmuscle‐invasive disease (80% vs 25.6%; P < .001), received multiple courses of intravesical therapy (32.5% vs 1.2%; P < .001), and had a more protracted course from diagnosis to aggressive treatment (median, 23 months vs 2 months; P < .001). An explicit review of claims data indicated that between 31.6% and 46.8% of the 6326 bladder cancer deaths identified in the SEER‐Medicare data potentially were avoidable, depending on the survivorship threshold chosen. Patients whose deaths potentially were avoidable more commonly presented with nonmuscle‐invasive disease (66.7% vs 24.7%; P < .0001) and lower grade disease (35.1% vs 15.1%; P < .0001).

CONCLUSIONS:

The greatest inroads into reducing death from bladder cancer likely hinge on earlier detection or improvement of systemic therapies. However, changing physician practice may translate into nontrivial reductions in bladder cancer mortality. Cancer 2009. © 2009 American Cancer Society.
Reducing bladder cancer deaths
Authors: Konety Badrinath R.
Published: January 2009

In patients with bladder cancer, earlier cystectomy or other aggressive therapy can save the lives of some patients with nonmuscle‐invasive disease. It is imperative to develop criteria for performing preemptive cystectomy in patients with nonmuscle‐invasive disease who have failed intravesical therapies.
A novel algorithm to improve pathologic stage prediction of clinically organ‐confined muscle‐invasive bladder cancer
Authors: Margel David , Harel Amir , Yossepowitch Ofer , Baniel Jack
Published: January 2009

BACKGROUND:

An algorithm was created to predict pathologic stage in patients with clinically organ‐confined muscle‐invasive bladder cancer.

METHODS:

The sample consisted of 133 consecutive patients scheduled to undergo cystectomy. To develop a tool to predict nonorgan‐confined disease before surgery, principal component analysis (PCA) was applied. Patients were stratified into a training set (n = 89) and a validation set (n = 44), and 7 parameters were evaluated: levels of carcinoembryonic antigen, cancer antigen (CA) 125, and carbohydrate antigen (CA) 19‐9; clinical stage; presence of hydronephrosis; presence of carcinoma in situ; and initial tumor size >3 cm. PCA was applied to the training set to determine the weight of each parameter. A PCA score was generated for each patient in the set, and a cutoff defining nonorgan‐confined disease was established. The accuracy of the cutoff was quantified by the area under the receiver operator characteristics curve (AUC). The model was then applied to the validation set without recalculation; the AUC and the positive and negative predictive values of the validation set were calculated.

RESULTS:

On pathologic evaluation, 71 patients (53%) were found to have organ‐confined tumors and 62 patients (47%) had extravesical disease. The AUC was 0.85 in the training group (95% confidence interval [95% CI], 0.71‐0.97) and 0.84 in the validation group (95% CI, 0.75‐0.93). The positive and negative predictive values in the validation group were 88% (95% CI, 71%‐96%) and 94% (95% CI, 71%‐99%), respectively.

CONCLUSIONS:

The newly devised, internally validated, algorithm was 85% accurate in predicting nonorgan‐confined bladder disease before cystectomy. Further external validation in a large cohort was recommended as still necessary. Cancer 2009. © 2009 American Cancer Society.
Comparison of low‐temperature processes for oil and coenzyme Q10 extraction from mackerel and herring
Authors: Laplante Serge , Souchet Nathalie , Bryl Piotr
Published: January 2009

Among the fat fish species available from Eastern Quebec (Canada), whole Atlantic mackerel (Scomber scombrus) and herring (Clupea harengus) represent abundant fishery resources which are currently under‐utilized. They have relatively high contents of oil and coenzyme Q10 (CoQ10) in their tissues, which could be valuable for nutraceutical applications. Therefore, two low‐temperature extraction processes were compared for the recovery of oil and CoQ10 from these resources, such as enzymatic hydrolysis using Protamex™ and supercritical CO2 (SCO2) using fish lyophilizates. The results revealed that highest yields of oil and CoQ10 were obtained using the enzymatic hydrolysis process with mackerel. Whatever the process used, CoQ10 concentrations were higher in herring oil, due mainly to a more selective extraction of CoQ10 over that of the oil. The highest CoQ10 recovery rates (extraction efficiencies) were obtained using the enzymatic hydrolysis process with both types of fish, but also the SCO2 process with herring under some conditions. For mackerel, the lower CoQ10 recovery rates obtained from the SCO2 process were explained by its more important matrix effect. An economic assessment of both processes revealed that the enzymatic hydrolysis extraction process would be the most promising for up‐scaling the recovery of oil and CoQ10 from these resources.
CEC with new monolithic stationary phase based on a fluorinated monomer, trifluoroethyl methacrylate
Authors: Yurtsever Arda , Saraçogˇlu Berna , Tuncel Ali
Published: January 2009

A new, fluorinated monolithic stationary phase for CEC was first synthesized by a single‐stage, thermally initiated copolymerization of a fluorinated monomer, 2,2,2‐trifluoroethyl methacrylate (TFEM) and ethylene dimethacrylate (EDMA) in the presence of a porogen mixture. In this preparation, 2‐acrylamido‐2‐methyl‐1‐propanesulfonic acid was used as the charge‐bearing monomer. The porogen mixture was prepared by mixing isoamylalcohol and 1,4‐butanediol. A clear increase in the electroosmotic mobility was observed with increasing pH. The electroosmotic mobility decreased with increasing ACN concentration. Poly(TFEM‐co‐EDMA) monolith prepared under optimized polymerization conditions was successfully used in the separation of alkylbenzenes and phenols by CEC. The best chromatographic separation for alkylbenzenes was performed with lower ACN concentrations (i.e. 60% v/v) with respect to the common acrylic‐based CEC monoliths. The theoretical plate numbers up to 220 000 plates/m were achieved in the reversed phase separation of phenols. Poly(TFEM‐co‐EDMA) monolith also allowed the simultaneous separation of aniline and benzoic acid derivatives by a single run and by using a lower ACN concentration in the mobile phase with respect to the similar electrochromatographic separations. A stable retention behaviour in reversed phase separation of alkylbenzenes was obtained with the poly(TFEM‐co‐EDMA) monolith.
Influence of different proteomic protocols on degree of high‐coverage identification of nonspecific lipid transfer protein 1 modified during malting
Authors: Chmelik Josef , Zidkova Jitka , Rehulka Pavel , Petry‐Podgorska Inga , Bobalova Janette
Published: January 2009

Both top‐down (combining protein separation with MS analysis of intact proteins) and bottom‐up (MS analysis of digested proteins) proteomic approaches were used for detailed characterization of nonspecific lipid transfer protein from barley malt. The aim was obtaining high‐coverage of the primary structure of the proteins and the determination of PTMs such as lipid adduction and glycation. Here we present an influence of 15 proteomic protocols (differing in applied separation technique, enzyme and digestion procedure) on the extent of the coverage of the protein primary structure. The most successful protocols were in‐gel digestion with trypsin of alkylated protein and in‐solution digestions with trypsin or trypsin/chymotrypsin mixture of the nonalkylated protein. Totally, full sequence coverage based on the PMF and 85% sequence coverage based on the peptide fragmentation including PTMs was obtained.
Chlorophyllin significantly reduces benzo[a]pyrene‐DNA adduct formation and alters cytochrome P4501A1and1B1expression and EROD activity in normal human mammary epithelial cells
Authors: Keshava Channa , Divi Rao L. , Einem Tracey L. , Richardson Diana L. , Leonard Sarah L. , Keshava Nagalakshmi , Poirier Miriam C. , Weston Ainsley
Published: January 2009

We hypothesized that chlorophyllin (CHLN) would reduce benzo[a]pyrene‐DNA (BP‐DNA) adduct levels. Using normal human mammary epithelial cells (NHMECs) exposed to 4 μM BP for 24 hr in the presence or absence of 5 μM CHLN, we measured BP‐DNA adducts by chemiluminescence immunoassay (CIA). The protocol included the following experimental groups: BP alone, BP given simultaneously with CHLN (BP+CHLN) for 24 hr, CHLN given for 24 hr followed by BP for 24 hr (preCHLN, postBP), and CHLN given for 48 hr with BP added for the last 24 hr (preCHLN, postBP+CHLN). Incubation with CHLN decreased BPdG levels in all groups, with 87% inhibition in the preCHLN, postBP+CHLN group. To examine metabolic mechanisms, we monitored expression by Affymetrix microarray (U133A), and found BP‐induced up‐regulation ofCYP1A1andCYP1B1expression, as well as up‐regulation of groups of interferon‐inducible, inflammation and signal transduction genes. Incubation of cells with CHLN and BP in any combination decreased expression of many of these genes. Using reverse transcription real time PCR (RT‐PCR) the maximal inhibition of BP‐induced gene expression, >85% forCYP1A1and >70% forCYP1B1, was observed in the preCHLN, postBP+CHLN group. To explore the relationship between transcription and enzyme activity, the ethoxyresorufin‐O‐deethylase (EROD) assay was used to measure the combined CYP1A1 and CYP1B1 activities. BP exposure caused the EROD levels to double, when compared with the unexposed controls. The CHLN‐exposed groups all showed EROD levels similar to the unexposed controls. Therefore, the addition of CHLN to BP‐exposed cells reduced BPdG formation andCYP1A1andCYP1B1expression, but EROD activity was not significantly reduced. Environ. Mol. Mutagen. 2009. Published 2009 Wiley‐Liss, Inc.
Molecular cytogenetic evaluation of the mechanism of micronuclei formation induced by camptothecin, topotecan, and irinotecan
Authors: Attia Sabry M. , Aleisa Abdulaziz M. , Bakheet Saleh A. , Al‐Yahya Abdulaziz A. , Al‐Rejaie Salim S. , Ashour Abdelkader E. , Al‐Shabanah Othman A.
Published: January 2009

We used the conventional bone marrow micronucleus test complemented with the fluorescent in situ hybridization with the minor satellite DNA probe to investigate the mechanisms of induction of micronuclei in mice treated with camptothecin and its clinical antineoplastic analogues topotecan and irinotecan. All experiments were performed with male Swiss albino mice. Single doses of 1 mg/kg camptothecin or 0.6 mg/kg topotecan were injected intraperitoneally and bone marrow was sampled at 30 hr (camptothecin) or 24 hr (topotecan) after treatment. A dose of 60 mg/kg irinotecan was injected intravenously, once every fourth day for 13 days and bone marrow was sampled 24 hr after the last treatment. In animals treated with camptothecin, a total of 1.07% micronuclei were found and 70% of them were centromere‐negative, indicating their formation by DNA strand breaks and reflecting the predominant clastogenic activity of camptothecin. Exposure to topotecan and irinotecan yielded 1.71 and 0.83% micronuclei, respectively. About 52.7 and 48.8% of the induced micronuclei, respectively, were centromere‐positive, indicating their formation by whole chromosomes and reflecting the aneugenic activity of both compounds. Correspondingly, about 47.3 and 51.2% of the induced micronuclei, respectively were centromere‐negative, demonstrating that topotecan and irinotecan not only induce chromosome loss but also DNA strand breaks. Both the clastogenic and aneugenic potential of these drugs can lead to the development of secondary tumors and abnormal reproductive outcomes. Therefore, the clinical use of these agents must be weighed against the risks of secondary malignancies in cured patients and persistent genetic damage of their potential offspring. Environ. Mol. Mutagen. 2009. © 2009 Wiley‐Liss, Inc.
Structural motifs modulating the carcinogenic risk of aromatic amines
Authors: Benigni Romualdo , Worth Andrew , Netzeva Tatiana , Jeliazkova Nina , Bossa Cecilia , Gruska Andreas , Franke Rainer
Published: January 2009

The structure alerts (SA) for carcinogenicity/mutagenicity are a repository of the science on chemical biological interactions; in addition, they have a crucial role in practical applications for risk assessment. In predictive toxicology, it is crucial that knowledge of SAs is accompanied by knowledge of the structural motifs that modulate their effects. Recently, we have compiled an updated list of SAs implemented in the expert system Toxtree 1.50 (open source, freely available). These SAs are aimed at discriminating between active and inactive chemicals, and include only modulating factors with a high probability of eliminating completely the effect of the SA. Here we have examined the factors that modulate carcinogenic potency: this is an additional piece of information that can have a role in fine‐tuning a risk assessment. The case study selected is the carcinogenic potential of the aromatic amines in rats and mice. As the carcinogenic potency of these compounds is different in mice and rats (correlation coefficient = 0.546), there are both agreements and differences in the pattern of these motifs. Differences are observed mainly for the motifs that decrease the carcinogenic potency of aromatic amines. In mice, substitutions ortho and meta to the amino group tend to decrease the potency, as well as −NO2in any position. In rats, these motifs affect the potency to a more limited extent. On the other hand, increasing effects are quite similar in the two animals and are exerted mainly by additional rings, tricyclic systems, five‐numbered rings, and N‐heteroaromatic systems. Environ. Mol. Mutagen. 2009. © 2009 Wiley‐Liss, Inc.
LSM‐YSZ Cathode with Infiltrated Cobalt Oxide and Cerium Oxide Nanoparticles
Authors: Imanishi N. , Ohno R. , Murata K. , Hirano A. , Takeda Y. , Yamamoto O. , Yamahara K.
Published: January 2009

To improve the La0.8Sr0.2MnO3 (LSM) –8 mol‐%Y2O3‐ZrO2 (YSZ) cathode performance of intermediate temperature solid oxide fuel cells (SOFC), Co3O4 and CeO2 nanoparticles were infiltrated into the cathode. Co‐infiltration of these oxide particles drastically enhanced the cell performance between 800 and 600 °C. The infiltrated CeO2 suppressed the aggregation of nanoparticle Co3O4, resulting in the high catalytic activity of the nanoparticle Co3O4 for oxygen reduction at intermediate temperatures. The anode‐supported SOFC with Co3O4 and CeO2 co‐infiltrated LSM‐YSZ cathode at 700 °C showed the high specific power density of 0.58 W cm–2 at 0.7 V.
Loss of 16q in high grade breast cancer is associated with estrogen receptor status: Evidence for progression in tumors with a luminal phenotype?
Authors: Natrajan Rachael , Lambros Maryou B. K. , Geyer Felipe C. , Marchio Caterina , Tan David S. P. , Vatcheva Radost , Shiu Kai‐Keen , Hungermann Daniela , Rodriguez‐Pinilla Socorro Maria , Palacios Jose , Ashworth Alan , Buerger Horst , Reis‐Filho Jorge S.
Published: January 2009

Loss of the long arm of chromosome 16 (16q) is observed in the vast majority of low grade/grade I (GI) invasive ductal carcinomas of no special type (IDC‐NSTs), whereas this event is uncommonly seen in high grade/grade III (GIII) IDC‐NSTs. Together with data on the pathology and genetics of breast cancer recurrences, this has led to the proposal that GI and GIII breast cancers evolve through distinct genetic pathways and that progression from GI to GIII is an unlikely biological phenomenon. We compared the genomic profiles of GIII‐IDC‐NSTs with 16q whole arm loss (16qWL) according to estrogen receptor (ER) status. 16qWL was found in 36.5% of cases and was significantly associated with ER expression and luminal phenotype. ER+ GIII‐IDC‐NSTs with 16qWL displayed significantly higher levels of genomic instability than ER+ IDC‐NSTs without 16qWL. Furthermore, ER+ and ER− IDC‐NSTs stratified according to the presence of 16qWL harbored distinct patterns of genetic aberrations. Interestingly, ER+/16qWL tumors displayed genetic features usually found in tumors with homologous DNA repair defects and significantly more frequently harbored heterozygous loss of BRCA2 than the remaining ER+ cancers. Our results demonstrate that approximately one third of GIII tumors harbor 16qWL, confirming that progression from low to high grade breast cancer is not found in the majority of breast cancers. 16qWL was significantly more prevalent in ER+/luminal GIII‐IDC‐NSTs. Given that GI breast cancers harbor a luminal phenotype, our results suggest that if progression from GI to GIII breast cancer does happen, it may preferentially occur in breast cancers of luminal phenotype. © 2009 Wiley‐Liss, Inc.
Prognosis is important in decisionmaking in Dutch nursing home patients with dementia and pneumonia
Authors: van der Steen Jenny T. , Helton Margaret R. , Ribbe Miel W.
Published: January 2009

Objective

To explore how physicians treating nursing home residents with dementia and pneumonia in the Netherlands consider prognosis in their treatment decision.

Methods

Survey study with data collected between July 2006 and March 2008. Physicians (n = 69) from 54 nursing homes in the Netherlands completed a questionnaire on symptoms, treatment, and prognosis for their next dementia patient newly diagnosed with pneumonia. They were also asked a general question regarding withholding antibiotic treatment and prognosis. Outcome was assessed at least two months afterwards. Two‐week mortality risk if treated with antibiotics was calculated with a validated prognostic score.

Results

The patients not treated with antibiotics had high (92%) actual 2‐week mortality while only 12% of patients treated with antibiotics died. Physicians believed that mortality risk was high in the untreated group and would have been only slightly lower if treated with antibiotics (mean estimated risk 73%), which was higher than predicted from the risk score (42%). In general, three‐quarters of physicians considered withholding antibiotics appropriate for mortality risks between 75% and 90%.

Conclusions

Prognosis is an important consideration when Dutch nursing home physicians make antibiotic treatment decisions for patients with dementia and pneumonia. This suggests they prefer not to treat with antibiotics when to do so is probably futile. Physicians in other countries may hold different views on futility, which should be addressed in larger, cross‐national comparative studies. Copyright © 2009 John Wiley & Sons, Ltd.
Effect of autologous platelet adhesives on dermal fat graft resorption following reconstruction of a superficial parotidectomy defect: A double‐blinded prospective trial
Authors: Chandarana Shamir , Fung Kevin , Franklin Jason H. , Kotylak Trevor , Matic Damir B. , Yoo John
Published: January 2009

Background.

Dermal fat grafts are used to reconstruct facial contour defects but may undergo variable resorption. Application of autologous platelet adhesive may improve outcomes. The primary objective was to compare resorption of dermal fat grafts for parotidectomy defects, between patients receiving autologous platelet adhesive versus controls.

Methods.

This was a double‐blinded prospective cohort at a tertiary care center. Volumetric analyses of dermal fat graft measured by MRI scans. Resorption was determined by comparing 1‐ and 6‐month MRIs in each patient. Complications, Frey's Syndrome, and patient satisfaction were also assessed.

Results.

Twelve patients completed the study. A significant reduction in graft resorption was seen in the treatment group (57% vs. 31%, p = .01). Three patients in the control group developed fat liquefaction. Patient perceived significant differences in scar and contour.

Conclusion.

Application of autologous platelet adhesives improved graft viability and patient satisfaction at 6 months. © 2009 Wiley Periodicals, Inc. Head Neck, 2009
Two novel CYP21A2 missense mutations in Italian patients with 21‐hydroxylase deficiency: Identification and functional characterisation
Authors: Concolino Paola , Vendittelli Francesca , Mello Enrica , Alinovi Cristiana Carelli , Minucci Angelo , Carrozza Cinzia , Santini Stefano Angelo , Zuppi Cecilia , Capoluongo Ettore
Published: January 2009

Steroid 21‐hydroxylase deficiency is present in more than 90% of patients with congenital adrenal hyperplasia (CAH), an inherited metabolic disorder of adrenal steroidogenesis. Impaired enzymatic activity leads to the accumulation of metabolic intermediates (progesterone and 17‐hydroxyprogesterone), which results in excessive androgen production and varied signs of virilisation. CYP21A2 is an active gene and encodes for the steroid 21‐hydroxylase enzyme, whereas CYP21A1P is an inactive pseudogene that contains a series of deleterious mutations. The major part of disease‐ causing mutations in CYP21A2 alleles are CYP21A1P‐derived sequence transferred to the active gene by macro or microconversion events. Only around 5% of all disease‐causing CYP21A2 alleles harbour rare mutations that do not originate from the pseudogene. In this report, we report the characterisation of two novel CYP21A2 missense mutations (p.H119R and p.I194N) found in two unrelated Italian patients with nonclassic (NC) CAH clinical diagnosis. Functional in vitro assays for mutagenized CYP21 enzymes were performed in transiently transfected mammalian cells to test the residual enzyme activity and the apparent kinetic values. The residual activities obtained for p.H119R and p.I194N mutants allowed to classify them as NC‐CAH associated mutations. These results correlate with the rate of severity of the patients' disease. Finally, the new p.H119R and p.I194N mutations should be included in the panel of those already listed for association with the NC form of 21‐hydroxylase deficiency. © 2009 IUBMB IUBMB Life 61(3): 229–235, 2009
AMP‐activated protein kinase suppresses LXR‐dependent sterol regulatory element‐binding protein‐1c transcription in rat hepatoma McA‐RH7777 cells
Authors: Yang Jian , Craddock Lauren , Hong Sharon , Liu Zhi‐Mei
Published: January 2009

AMP‐activated protein kinase (AMPK) is an intracellular fuel sensor that plays a key role in regulating fatty acid synthesis in liver. Sterol regulatory element‐binding protein (SREBP)‐1c is a master regulator of hepatic lipogenic gene expression. It has long been documented that AMPK activation suppresses hepatic SREBP‐1 mRNA and nuclear SREBP‐1 protein. But the mechanism remains undefined. In this study we investigated the molecular mechanisms by which AMPK downregulates hepatic SREBP‐1c mRNA using a novel model cell line McA‐RH7777. We found that AMPK is robustly activated in rat hepatoma McA‐RH7777 cells treated with two widely used AMPK activators, AICAR and metformin, and AMPK activation sharply suppresses SREBP‐1c mRNA and nuclear SREBP‐1c protein, but not SREBP‐1a mRNA derived from the same gene. These inhibitory effects are reversed by the AMPK inhibitor Compound C or 8‐BrAMP, demonstrating the requirement of AMPK in the suppression of SREBP‐1c mRNA and nuclear SREBP‐1c protein by AICAR and metformin. AMPK does not enhance SREBP‐1c mRNA degradation in the presence of the general transcription inhibitor actinomycin D; instead it inhibits SREBP‐1c promoter activity in a luciferase reporter assay. AMPK‐mediated inhibition of SREBP‐1c promoter activity can also be abrogated by the AMPK inhibitor Compound C. Furthermore AMPK activation significantly attenuates the synthetic liver X receptor (LXR) ligand T0901317‐induced SREBP‐1c promoter activity. AMPK also inhibits cleavage of LXR ligand‐induced SREBP‐1c precursor. We conclude that AMPK suppresses hepatic SREBP‐1c mRNA expression by inhibiting LXR‐dependent SREBP‐1c transcription via inhibition of endogenous LXR ligand production and by inhibiting SREBP‐1c processing in McA‐RH7777 cells. J. Cell. Biochem. 106: 414–426, 2009. © 2009 Wiley‐Liss, Inc.
Primary care mental health: a new frontier for psychology
Authors: Gunn, William B. , Blount Alexander
Published: January 2009

The medical system in this country is divided into primary care and specialty care. Mental health is for the most part a specialty service dependent on referrals, often from primary care providers. The authors propose a new model where psychologists work in collaboration with primary care medical teams. This integrated, coordinated model enables psychologists to help patients they would not otherwise see in a mental health system. Examples of patients in this category are seniors, those with somatizing disorders, and those experiencing the challenges of dealing with a chronic illness. This model also enables psychologists to provide consultation to the medical teams. In this article, the authors discuss the world of the primary care medical team and present the rationale for integration or collaboration. They describe the barriers to collaborative practices and ways to overcome these barriers. Finally, they present practical strategies that psychologists can use on a regular basis to increase their collaboration with primary care. These strategies can be used by those who work in colocated practices as well as those who work in separate locations. © 2009 Wiley Periodicals, Inc. J Clin Psychol 65:1–18, 2009.
Impact of behavioral health consultant interventions on patient symptoms and functioning in an integrated family medicine Clinic
Authors: Bryan Craig J. , Morrow Chad , Appolonio Kathryn Kanzler
Published: January 2009

Patterns of symptomatic and functional change associated with behavioral health consultant (BHC) intervention in an integrated family medicine clinic were investigated among 338 primary care patients under routine conditions without exclusion. Patients were referred to the BHC by primary care providers (PCPs) and participated in one to four brief, behaviorally oriented appointments in primary care. The Behavioral Health Measure‐20 (BHM) was completed at each appointment. Results indicated that higher levels of distress at baseline were associated with more follow‐up appointments, and that patients demonstrated simultaneous, clinically meaningful improvement in well‐being, symptoms, and functioning in as few as two to three BHC appointments. Patterns of clinical improvement support the effectiveness of BHC interventions, but contradict the phase model of psychotherapy (Howard, Lueger, Maling, & Martinovich, 1993). © 2009 Wiley Periodicals, Inc. J Clin Psychol: In Session 65:1–13, 2009.
Coping mediates outcome following a randomized group intervention for HIV‐positive bereaved individuals
Authors: Smith Nathan Grant , Tarakeshwar Nalini , Hansen Nathan B. , Kochman Arlene , Sikkema Kathleen J.
Published: January 2009

The purpose of this study was to examine the mechanisms responsible for the beneficial psychological effects of a coping‐focused group intervention for HIV‐positive individuals who had lost loved ones to AIDS. Data from 235 HIV‐positive men and women enrolled in a randomized controlled clinical trial testing a coping‐focused group intervention were analyzed using a multiple‐indicator‐multiple‐cause (MIMIC) structural equation model. Results revealed that the effects of the intervention on decreases in depression and grief were mediated by decreases in avoidant coping. Specifically, participants in the intervention condition decreased their use of avoidant coping. Decreases in avoidant coping, in turn, were related to decreased depression and grief. The results of this study help to validate the use of coping‐focused interventions for HIV‐positive bereaved individuals. © 2009 Wiley Periodicals, Inc. J Clin Psychol: In Session 65:1–17, 2009.
Using behavioral health consultants to treat insomnia in primary care: a clinical case series
Authors: Goodie Jeffrey L. , Isler William C. , Hunter Christopher , Peterson Alan L.
Published: January 2009

Cognitive–behavioral treatments for insomnia are as effective as medications and have longer lasting effects. The current study used a clinical case series design to evaluate the effectiveness of a brief behavioral intervention for insomnia delivered in a nonresearch, real‐world family medicine clinical setting. Participants included 29 sleep‐impaired patients who were seen regardless of their comorbid conditions. The treatment included three brief visits with a behavioral health consultant (BHC), plus the provision of a self‐help insomnia‐treatment book. At posttreatment 83% of participants achieved a mean sleep efficiency >85%, as compared to only 14% at baseline. Limited‐contact behavioral treatment of insomnia delivered by BHCs within a collaborative care family medicine clinic effectively reduced symptoms of insomnia, regardless of comorbid medical diagnoses. © 2009 Wiley Periodicals, Inc. J Clin Psychol: In Session, 65:1–11, 2009.
Long‐term follow‐up of patients with hepatitis C with a normal alanine aminotransferase
Authors: Kumada Takashi , Toyoda Hidenori , Kiriyama Seiki , Sone Yasuhiro , Tanikawa Makoto , Hisanaga Yasuhiro , Kanamori Akira , Atsumi Hiroyuki , Takagi Makiko , Nakano Satoshi , Arakawa Takahiro , Fujimori Masashi
Published: January 2009

An attempt was made to identify factors influencing the cumulative probability of an increased alanine aminotransferase (ALT) level and hepatocarcinogenesis in hepatitis C patients with a normal ALT level initially. A total of 398 consecutive patients with a normal ALT level initially for 6 months or more and follow‐up period longer than 3 years during the period January 1995 to December 2004 were included. Patients were classified by ALT level into three groups: Group A (3–20 IU/L), Group B (21–30 IU/L), and Group C (31–35 IU/L). Factors associated with the cumulative probability of increased ALT level and hepotocarcinogenesis were evaluated. Women in groups B and C and men in Group C showed high cumulative probabilities of increased ALT levels. Factors associated with increased ALT were a high ALT level (Group B, relative risk; 1.758 [95% confidence interval: 1.290–2.392], P < 0.001, Group C, 3.328 [2.256–4.909], P < 0.001), high lactate dehydrogenase level (2.352 [1.445–3.829], P = 0.001), or low total cholesterol level (1.957 [1.330–2.882], P = 0.001). Factors associated with incidence of hepatocellular carcinoma were increased age (3.088 [1.025–9.308], P = 0.045), high ALT level (Group C, 5.803 [1.530–22.066], P = 0.010), and high total bilirubin level (8.309 [2.235–30.888], P = 0.002). In patients with hepatitis C with a normal ALT level initially, an ALT level of 21–35 IU/L is a risk factor for an increased ALT level and hepatocarcinogenesis. J. Med. Virol. 81:446–451, 2009. © 2009 Wiley‐Liss, Inc.
Prevalence and type distribution of high‐risk human papillomavirus infection in women undergoing voluntary cervical cancer screening in Italy
Authors: Agarossi Alberto , Ferrazzi Enrico , Parazzini Fabio , Perno Carlo Federico , Ghisoni Luciano
Published: January 2009

The aim of this survey was to assess the prevalence and distribution of oncogenic human papillomavirus (HPV) genotypes in women who underwent screening for cervical cancer in Italy. The correlation of genotypes with the cytological results was also evaluated. Cervical samples were collected from 9,947 self‐referring women for cervical cancer screening. Participants were screened by liquid‐based cytology and high‐risk HPV testing using the Hybrid Capture 2 test. Positive samples were genotyped by PCR. Samples (1,474; 14.8%) were positive for high‐risk HPV. The prevalence was 29.4% in the 15–19 years‐group, decreasing progressively to 6.1% at 50–54 years of age and increasing to 12.2% in those aged over 65 years. HPV 16 was the genotype detected most frequently followed by HPV 31, HPV 18, HPV 56, and HPV 51. HPV 16 or 18 were present in 4% of women with normal cytology and both were detected contemporarily in only 14 women. Twenty‐two percent of atypical squamous cells, 26% of low‐grade and 56% of high‐grade squamous intraepithelial lesions at cytology were positive for HPV 16 and/or 18. The prevalence of HPV infection in Italy is in agreement with that reported worldwide. HPV 16 was the prevalent genotype. The concomitant infection with HPV 16 and HPV 18 (vaccine targets) was found rarely. Apart from HPV 16 and 18, there was a substantial presence of HPV genotypes against which the vaccines available currently have shown cross‐protection efficacy. The findings of this study may contribute to reliable predictions on the potential efficacy of an HPV vaccine in clinical practice. J. Med. Virol. 81:529–535, 2009. © 2009 Wiley‐Liss, Inc.
Successful valganciclovir treatment of post‐transplant cytomegalovirus infection in the presence of UL97 mutation N597D
Authors: Iwasenko Jenna M. , Scott Gillian M. , Rawlinson William D. , Keogh Anne , Mitchell Daniel , Chou Sunwen
Published: January 2009

Mutations in the human cytomegalovirus (CMV) UL97 protein kinase are the most common mechanism of ganciclovir (GCV) resistance in the clinical setting. A CMV strain with a previously unrecognized UL97 mutation N597D was identified in the blood of a heart transplant recipient who experienced a persistent CMV infection with high viral loads accompanying pain and fever while receiving valganciclovir (valGCV) therapy. The N597D mutation was transferred by mutagenesis to an antiviral sensitive CMV strain for analysis of antiviral susceptibility by standardized phenotypic assay. Recombinant phenotyping showed N597D conferred a less than twofold increase in GCV IC50 compared to the sensitive control strain. Despite the presence of this mutation, valGCV eventually resolved the infection after 6 weeks of therapy. A subsequent CMV reactivation was also responsive to valganciclovir. This case illustrates the diversity of UL97 mutations in the codon segment 590–607 usually associated with GCV resistance, with some mutations producing minimal levels of resistance that do not preclude a therapeutic response to the drug. Accurate interpretation of genotypic test results ultimately requires experimental determination of the level of resistance conferred by newly discovered UL97 mutations. J. Med. Virol. 81:507–510, 2009. © 2009 Wiley‐Liss, Inc.
Analysis of gene transcription in sera during chronic hepatitis C infection
Authors: Carpentier Arnaud , Conti Filoména , Carrière Matthieu , Aoudjehane Lynda , Miroux Céline , Moralès Olivier , Calmus Yvon , Groux Hervé , Auriault Claude , Pancré Véronique , Delhem Nadira , Podevin Philippe
Published: January 2009

Alternative, non‐invasive techniques are necessary to monitor the progression of liver disease during chronic hepatitis C. Firstly, because serum is the most accessible material for studies using qPCR in microplates, gene transcription was compared in 219 selected genes involved in the pathogenesis of hepatitis C virus (HCV) infection between sera, PBMCs and liver samples collected simultaneously from five patients infected chronically. Secondly, using sera, gene profiles were compared between HCV‐infected patients (n = 10) and healthy controls (n = 10). In addition, the influence of alcohol intake was examined in patients infected with HCV genotype‐1. Firstly, amplifiable mRNAs were obtained in all samples. After amplification, significant correlations were observed between: liver versus serum; liver versus PBMCs; and serum versus PBMCs (r2 = 0.37, r2 = 0.54, r2 = 0.49, respectively). A comparison of gene transcription by gene involved in T‐ and B‐cell markers, adhesion molecules, apoptosis, liver matrix turnover and inflammation, revealed comparable, significant correlations between serum and liver, (r2 = 0.30, r2 = 0.60, r2 = 0.51, r2 = 0.51, r2 = 0.26, and r2 = 0.61 respectively). Secondly, a quantitative analysis of gene expression in sera between genotype‐1b‐infected patients and healthy controls revealed that 41 genes involved closely in T‐cell activation and apoptosis were over‐expressed significantly in patients infected with HCV. In these patients, alcohol consumption was associated with an increased expression of six genes involved in the inflammatory response, together with a decrease of genes associated with dendritic cell function. It is concluded that in patients infected with HCV, serum can be used to evaluate expression of liver genes. Further prospective studies are clearly needed to validate the initial results and to define the relevant genes. J. Med. Virol. 81:473–480, 2009. © 2009 Wiley‐Liss, Inc.
A matched case‐controlled study of 48 and 72 weeks of peginterferon plus ribavirin combination therapy in patients infected with HCV genotype 1b in Japan: amino acid substitutions in HCV core region as predictor of sustained virological response
Authors: Akuta Norio , Suzuki Fumitaka , Hirakawa Miharu , Kawamura Yusuke , Yatsuji Hiromi , Sezaki Hitomi , Suzuki Yoshiyuki , Hosaka Tetsuya , Kobayashi Masahiro , Kobayashi Mariko , Saitoh Satoshi , Arase Yasuji , Ikeda Kenji , Kumada Hiromitsu
Published: January 2009

Substitution of amino acid (aa) 70 and 91 in the core region of HCV genotype 1b is a useful pretreatment predictor of efficacy of 48‐week peginterferon (PEG‐IFN) plus ribavirin (RBV) therapy. Here, we determined the efficacy of 72‐week PEG‐IFN/RBV and the predictive factors to such therapy in a case–control study matched for sex, age, and periods from the start of treatment to initial point of HCV RNA‐negative. We compared the treatment efficacy of 72‐week regimen in 65 patients with that of 48‐week in 130 patients, who were infected with HCV genotype 1b and treated with PEG‐IFN/RBV. They consisted mainly of late virological responders (LVR) (HCV RNA‐positive at 12 weeks and negative at 24 weeks after start of treatment). Sustained virological response (SVR) was achieved by 61.5% and 32.3% of patients of the 72‐ and 48‐week groups, respectively, while non‐virological response was noted in 9.2% and 29.2% of the respective groups. Multivariate analysis identified substitution of aa 70 and 91 (Arg70 and/or Leu91) and duration of treatment (72‐week) as independent parameters that significantly influenced SVR. For Arg70 and/or Leu91 of core region, SVR rate was significantly higher in 72‐ (68.0%) than 48‐week group (37.8%). For wild‐type of ISDR, SVR rate was significantly higher in 72‐ (61.2%) than in 48‐week group (29.3%). We conclude that 72‐week PEG‐IFN/RBV improves SVR rate for LVR, especially those with Arg70 and/or Leu91 of core region or wild‐type of ISDR. Substitution of aa 70 and 91 is also a useful pretreatment predictor of response to 72‐week PEG‐IFN/RBV. J. Med. Virol. 81:452–458, 2009. © 2009 Wiley‐Liss, Inc.
Susceptibility of hepatitis B virus to lamivudine restored by resistance to adefovir
Authors: Zaaijer H.L. , Takkenberg R.B. , Weegink C.J. , Rebers S.P.H. , Menting S. , Reesink H.W. , Schinkel J. , Molenkamp R.
Published: January 2009

Serial monotherapy and add‐on regimes for treatment of chronic hepatitis B virus (HBV) infection may induce the accumulation of viral resistance mutations in patients, reducing the options for ongoing viral suppression. The induction of antiviral resistance by serial application of polymerase inhibitors does not necessarily imply that the subsequent combined use of the drugs will fail. Some HIV strains resistant to one polymerase inhibitor show increased susceptibility to another polymerase inhibitor. After failure of sequential lamivudine and adefovir monotherapy, two patients with hepatitis B changed to treatment with lamivudine plus adefovir and had renewed suppression of HBV. To study the mutational history of resistant HBV subpopulations in the two patients, a part of the HBV polymerase gene was amplified, cloned, sequenced, and analyzed for the presence of mutations, in sequential plasma samples. In both patients serial monotherapy caused the replacement in all HBV clones of wild‐type virus by classical lamivudine resistant mutants (L180M and M204V/I), which were replaced subsequently by adefovir resistant mutants (A181V and N236T). When finally lamivudine was added to adefovir, the A181V adefovir mutation persisted in all clones and lamivudine‐related mutations did not reappear. During 18 months of combination therapy, HBV‐DNA levels decreased 10,000, respectively, 1,000‐fold, despite the earlier resistance to lamivudine and adefovir. Although clinically insufficient, this effect indicates that HBV polymerase resistance mutations may be antagonistic, which is relevant if chronic HBV infection is to be treated by a combination of polymerase inhibitors. J. Med. Virol. 81:413–416, 2009. © 2009 Wiley‐Liss, Inc.