Case report: Initial atypical skeletal symptoms and dental anomalies as first signs of Gardner syndrome: the importance of genetic analysis in the early diagnosis.

Background: Gardner syndrome is a rare genetic cancer predisposition disorder characterized by intestinal polyposis, multiple osteomas, and soft and hard tissue tumors. Dental anomalies are present in approximately 30%-70% of patients with Gardner syndrome and can be discovered during routine dental examinations. However, sometimes the diagnosis is challenging due to the high clinical variability and incomplete clinical picture. Herein, we report a family with various dental and bone anomalies, in which the definitive diagnosis was established with the help of a comprehensive genetic analysis based on state-of-the-art next-generation sequencing technology. Case presentation: A 17-year-old female index patient presented with dental (caries, impacted, retained and anteriorly located teeth) and atypical bone anomalies not resembling Gardner syndrome. She was first referred to our Genetic Counselling Unit at the age of 11 due to an atypical bone abnormality identified by a panoramic X-ray. Tooth 3.6 was surgically removed and the histopathology report revealed a Paget's disease-like bone metabolic disorder with mixed osteoblastic and osteoclastic activity of the mandible. A small lumbar subcutaneous tumor was discovered by physical examination. Ultrasound examination of the tumor raised the possibility of a soft tissue propagation of chondromatosis. Her sister, 2 years younger at the age of 14, had some benign tumors (multiple exostoses, odontomas, epidermoid cysts) and impacted teeth. Their mother had also skeletal symptoms. Her lower teeth did not develop, the 9th-10th ribs were fused, and she complained of intermittent jaw pain. A cranial CT scan showed fibrous dysplasia on the cranial bones. Whole exome sequencing identified a heterozygous pathogenic nonsense mutation (c.4700C>G; p.Ser1567*) in the APC gene in the index patient's DNA. Targeted sequencing revealed the same variant in the DNA of the other affected family members (the sister and the mother). Conclusion: Early diagnosis of this rare, genetically determined syndrome is very important, because of the potentially high malignant transformation of intestinal polyps. Dentists should be familiar with the typical maxillofacial features of this disorder, to be able to refer patients to genetic counseling. Dental anomalies often precede the intestinal polyposis and facilitate the early diagnosis, thereby increasing the patients' chances of survival. Genetic analysis may be necessary in patients with atypical phenotypic signs.

NADPH Oxidases Connecting Fatty Liver Disease, Insulin Resistance and Type 2 Diabetes: Current Knowledge and Therapeutic Outlook.

Nonalcoholic fatty liver disease (NAFLD), characterized by ectopic fat accumulation in hepatocytes, is closely linked to insulin resistance and is the most frequent complication of type 2 diabetes mellitus (T2DM). One of the features connecting NAFLD, insulin resistance and T2DM is cellular oxidative stress. Oxidative stress refers to a redox imbalance due to an inequity between the capacity of production and the elimination of reactive oxygen species (ROS). One of the major cellular ROS sources is NADPH oxidase enzymes (NOX-es). In physiological conditions, NOX-es produce ROS purposefully in a timely and spatially regulated manner and are crucial regulators of various cellular events linked to metabolism, receptor signal transmission, proliferation and apoptosis. In contrast, dysregulated NOX-derived ROS production is related to the onset of diverse pathologies. This review provides a synopsis of current knowledge concerning NOX enzymes as connective elements between NAFLD, insulin resistance and T2DM and weighs their potential relevance as pharmacological targets to alleviate fatty liver disease.

NADPH Oxidase 4 (NOX4) in Cancer: Linking Redox Signals to Oncogenic Metabolic Adaptation.

Cancer cells can survive and maintain their high proliferation rate in spite of their hypoxic environment by deploying a variety of adaptative mechanisms, one of them being the reorientation of cellular metabolism. A key aspect of this metabolic rewiring is the promotion of the synthesis of antioxidant molecules in order to counter-balance the hypoxia-related elevation of reactive oxygen species (ROS) production and thus combat the onset of cellular oxidative stress. However, opposite to their negative role in the inception of oxidative stress, ROS are also key modulatory components of physiological cellular metabolism. One of the major physiological cellular ROS sources is the NADPH oxidase enzymes (NOX-es). Indeed, NOX-es produce ROS in a tightly regulated manner and control a variety of cellular processes. By contrast, pathologically elevated and unbridled NOX-derived ROS production is linked to diverse cancerogenic processes. In this respect, NOX4, one of the members of the NOX family enzymes, is of particular interest. In fact, NOX4 is closely linked to hypoxia-related signaling and is a regulator of diverse metabolic processes. Furthermore, NOX4 expression and function are altered in a variety of malignancies. The aim of this review is to provide a synopsis of our current knowledge concerning NOX4-related processes in the oncogenic metabolic adaptation of cancer cells.

Prevalence and correlation of severe early childhood caries among preschool children in Pécs / A súlyos korai gyermekkori fogszuvasodás elofordulása és összefüggései pécsi bölcsodések között.

Összefoglaló. Bevezetés és célkituzés: A 3 év alatti korcsoportban a súlyos kisgyermekkori fogszuvasodás (S-ECC) gyakoriságának vizsgálata és a kisgyermekes szülok szájegészséggel kapcsolatos ismereteinek felmérése. Módszer: Háromszázhatvankét, 36 hónapos vagy annál fiatalabb gyermek (átlagéletkor: 28,49 ± 5,25 hónap) fogászati szurovizsgálatát végeztük el, és szüleik számára önkéntesen kitöltheto kérdoívet állítottunk össze. A 306 kitöltött kérdoív eredményét a gyermekek fogászati statusával egyénenként összevetve statisztikai analízist végeztünk. Eredmények: A gyermekek fogászati szurésekor 15,46% volt a cariesprevalencia: df-t-index = 0,685 ± 2,20; az általunk javasolt, az elotört fogak számával módosított df-t-index = 0,758 ± 2,42, SiC-index = 2,06 ± 3,33. A vizsgált populációban nem találtunk tömött vagy fogszuvasodás miatt eltávolított fogat. A korábban szájhigiénés tájékoztatásban részesült/nem részesült szülok gyermekeinél nem volt szignifikáns különbség a szuvas fogak számának (p = 0,196), a fogyasztott folyadék cukortartalmának (81,5%/71,5%) és a bevitel módjának (p = 0,453) tekintetében. A gyermeküket 6 hónapos korukig kizárólagosan anyatejjel tápláló édesanyák nagyobb eséllyel választották a vízzel történo itatást (75%/52%) pohárból (68,1%/28,8%) a késobbiekben. Az édesanya legmagasabb iskolai végzettsége kulcsszerepet játszik mind az anyatejes táplálás melletti elhivatottságban (felsofok: 53,4%, középfok: 34,2%, alapfok: 37,5%), mind a cariesfrekvencia csökkentésében (p = 0,015). Következtetés: A szüloi szájhigiénés prevenciós tájékoztatás jelenleg nem hatékony. Eredményeink alapján a fogászati szurést 1 éves kor elott meg kell kezdeni, amely a hatékony, rendszeres fogászati prevenciós tanácsadás lehetoségét is megteremtené. Szükségesnek látjuk a védonok és a gyermekháziorvosok folyamatos továbbképzésében a kisgyermekkori fogszuvasodás alapismereteinek integrációját. Orv Hetil. 2021; 162(22): 861-869. INTRODUCTION AND OBJECTIVE: To investigate the frequency of severe early childhood caries (S-ECC) under 3 years of age and to assess the oral health related knowledge of parents/guardians with preschool children. METHOD: 362 children younger than 36 months (mean age: 28.49 ± 5.25 months) were screened and a voluntary questionnaire for their parents was compiled. Statistical analysis was carried out comparing the results of the 306 completed questionnaires with the dental status of the screened children. RESULTS: Caries prevalence in the examined population was 15.46%, df-t index = 0.685 ± 2.20, our suggested modified df-t index based on the number of erupted teeth = 0.758 ± 2.42, SiC-index = 2.06 ± 3.33. No filled or extracted tooth due to caries was found in the study group. There was no significant difference in the number of carious teeth (p = 0.196), consumption of sugar-containing drinks (81.5%/71.5%) and administration of drinks (p = 0.453) in the case of children whose parents had previously received/not received oral hygiene information. Mothers who exclusively breastfed until the age of 6 months were more likely to choose to offer water (75%/52%) from cup (68.1%/28.8%) later. The mother's highest level of education plays a key role in both the commitment to breastfeeding (tertiary: 53.4%, secondary: 34.2%, primary: 37.5%) and in reducing the incidence of caries (p = 0.015). CONCLUSION: Parental oral hygiene preventive instruction is currently ineffective. Based on our results, dental screening should be started before the age of 1 year, that would also provide an opportunity of effective, regular dental prevention counseling. There is a need for the integration of the basics of early childhood caries in the continuous professional training of district nurses and pediatricians. Orv Hetil. 2021; 162(22): 861-869.

Course induced dexterity development and cerebellar grey matter growth of dentistry students: a randomised trial.

This study primarily focuses on the assessment of dentistry students' improvement of manual skills resulting from their participation in courses. We aimed to prove that systematic manual skills development significantly improves dexterity. We hypothesized that the dexterity training regimen improves manual dexterity demonstrated by the HAM-Man (Hamburg Assessment Test for Medicine-Manual Dexterity) test scores and CGM (cerebellar grey matter) growth. Thirty volunteers were randomly divided into two equal groups (study and control). Firstly, volunteers were examined by the HAM-Man test and baseline MRI scans. Afterwards, a manual skills development course was launched for the "study group". Secondly, all the manual skills of the students were evaluated longitudinally, by the HAM-Man test. Simultaneously, the follow-up MRI scans were taken to observe morphologic changes in the cerebellum. The Wilcoxon signed-rank test and Student Paired t-test were used for statistical analyses. Value p < 0.05 was considered significant. After the training, significant growth of CGM as well as improvement on manual skill assessment tests, were found in the study group. Training courses are suitable for preparing students with low levels of dexterity for performing demanding tasks. The improvement is demonstrable by a wire bending test and by bilateral CGM enlargement as well.

[Objective measurement of manual dexterity of Parkinson patients operated with DBS]. / DBS-mûtéttel kezelt Parkinson-kóros betegek kézügyesség-javulásának objektív mérése.

The evaluation of hand dexterity is an important marker for the success of DBS (deep brain stimulation) operation in patients with Parkinson's disease. In this study we applied a simple, semiquantitative optical dental plaque staining technique for the evaluation of the hand dexterity. Ten patient with Parkinson's disease were involved in the study. After dental students aided tooth brushing, bacterial dental deposits (plaque) were stained then photographed, and quantified under standard conditions before and after DBS surgery. Our results showed a significant decrease in dental plaque deposits after DBS operation. This simple technique seems to be a routinely applicable marker for the evaluation of the hand dexterity. Our future plans is repeating the previous experiement on a higher number of cases.

Dentofacial mini- and microesthetics as perceived by dental students: A cross-sectional multi-site study.

BACKGROUND: How dental education influences students' dental and dentofacial esthetic perception has been studied for some time, given the importance of esthetics in dentistry. However, no study before has studied this question in a large sample of students from all grades of dental school. This study sought to fill that gap. The aim was to assess if students' dentofacial esthetic autoperception and heteroperception are associated with their actual stage of studies (grade) and if autoperception has any effect on heteroperception. METHODS: Between October 2018 and August 2019, a questionnaire was distributed to 919 dental students of all 5 grades of dental school at all four dental schools in Hungary. The questionnaire consisted of the following parts (see also the supplementary material): 1. Demographic data (3 items), Self-Esthetics I (11 multiple- choice items regarding the respondents' perception of their own dentofacial esthetics), Self-Esthetics II (6 Likert-type items regarding the respondents' perception of their own dentofacial esthetics), and Image rating (10 items, 5 images each, of which the respondents have to choose the one they find the most attractive). Both the self-esthetics and the photo rating items were aimed at the assessment of mini- and microesthetic features. RESULTS: The response rate was 93.7% (861 students). The self-perception of the respondents was highly favorable, regardless of grade or gender. Grade and heteroperception were significantly associated regarding maxillary midline shift (p < 0.01) and the relative visibility of the arches behind the lips (p < 0.01). Detailed analysis showed a characteristic pattern of preference changes across grades for both esthetic aspects. The third year of studies appeared to be a dividing line in both cases, after which a real preference order was established. Association between autoperception and heteroperception could not be verified for statistical reasons. CONCLUSION: Our findings corroborate the results of most previous studies regarding the effect of dental education on the dentofacial esthetic perception of students. We have shown that the effect can be demonstrated on the grade level, which we attribute to the specific curricular contents. We found no gender effect, which, in the light of the literature, suggests that the gender effect in dentofacial esthetic perception is highly culture dependent. The results allow no conclusion regarding the relation between autoperception and heteroperception.

Beta-Cell-Specific Expression of Nicotinamide Adenine Dinucleotide Phosphate Oxidase 5 Aggravates High-Fat Diet-Induced Impairment of Islet Insulin Secretion in Mice.

Aims: Nicotinamide adenine dinucleotide phosphate oxidases (NOX-es) produce reactive oxygen species and modulate ß-cell insulin secretion. Islets of type 2 diabetic subjects present elevated expression of NOX5. Here, we sought to characterize regulation of NOX5 expression in human islets in vitro and to uncover the relevance of NOX5 in islet function in vivo using a novel mouse model expressing NOX5 in doxycycline-inducible, ß-cell-specific manner (RIP/rtTA/NOX5 mice). Results:In situ hybridization and immunohistochemistry employed on pancreatic sections demonstrated NOX5 messenger ribonucleic acid (mRNA) and protein expressions in human islets. In cultures of dispersed islets, NOX5 protein was observed in somatostatin-positive (δ) cells in basal (2.8 mM glucose) conditions. Small interfering ribonucleic acid (siRNA)-mediated knockdown of NOX5 in human islets cultured in basal glucose concentrations resulted in diminished glucose-induced insulin secretion (GIIS) in vitro. However, when islets were preincubated in high (16.7 mM) glucose media for 12 h, NOX5 appeared also in insulin-positive (ß) cells. In vivo, mice with ß-cell NOX5 expression developed aggravated impairment of GIIS compared with control mice when challenged with 14 weeks of high-fat diet. Similarly, in vitro palmitate preincubation resulted in more severe reduction of insulin release in islets of RIP/rtTA/NOX5 mice compared with their control littermates. Decreased insulin secretion was most distinct in response to theophylline stimulation, suggesting impaired cyclic adenosine monophosphate (cAMP)-mediated signaling due to increased phosphodiesterase activation. Innovation and Conclusions: Our data provide the first insight into the complex regulation and function of NOX5 in islets implying an important role for NOX5 in δ-cell-mediated intraislet crosstalk in physiological circumstances but also identifying it as an aggravating factor in ß-cell failure in diabetic conditions.

H2O2 Metabolism in Normal Thyroid Cells and in Thyroid Tumorigenesis: Focus on NADPH Oxidases.

Thyroid hormone synthesis requires adequate hydrogen peroxide (H2O2) production that is utilized as an oxidative agent during the synthesis of thyroxin (T4) and triiodothyronine (T3). Thyroid H2O2 is generated by a member of the family of NADPH oxidase enzymes (NOX-es), termed dual oxidase 2 (DUOX2). NOX/DUOX enzymes produce reactive oxygen species (ROS) as their unique enzymatic activity in a timely and spatially regulated manner and therefore, are important regulators of diverse physiological processes. By contrast, dysfunctional NOX/DUOX-derived ROS production is associated with pathological conditions. Inappropriate DUOX2-generated H2O2 production results in thyroid hypofunction in rodent models. Recent studies also indicate that ROS improperly released by NOX4, another member of the NOX family, are involved in thyroid carcinogenesis. This review focuses on the current knowledge concerning the redox regulation of thyroid hormonogenesis and cancer development with a specific emphasis on the NOX and DUOX enzymes in these processes.

Comparison of fractionation strategies for offline two-dimensional liquid chromatography tandem mass spectrometry analysis of proteins from mouse adipose tissue.

In the frame of protein identification from mouse adipose tissue, two strategies were compared for the offline elution of peptides from a strong cation exchange (SCX) column in two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) analyses. First, the salt gradient (using K(+) as displacing agent) was evaluated from 25 to 500mM KCl. Then, a less investigated elution mode using a pH gradient (using citric acid and ammonium hydroxide) was carried out from pH 2.5 to 9.0. Equal amounts of peptide digest derived from mouse adipose tissue were loaded onto the SCX column and fractionated according to the two approaches. A total of 15 fractions were collected in two independent experiments for each SCX elution strategy. Then, each fraction was analyzed on a nanoLC-MS/MS platform equipped with a column-switching unit for desalting and enrichment. No substantial differences in peptide quality characteristics (molecular weight, isoelectric point, or GRAVY [grand average of hydropathicity] index distributions) were observed between the two datasets. The pH gradient approach was found to be superior, with 27.5% more unique peptide identifications and 10% more distinct protein identifications compared with the salt-based elution method. In conclusion, our data imply that the pH gradient SCX fractionation is more desirable for proteomics analysis of entire adipose tissue.

Triple combination therapy in hypertension: the antihypertensive efficacy of treatment with perindopril, amlodipine, and indapamide SR.

BACKGROUND AND OBJECTIVE: The blood pressure (BP) of most patients on antihypertensive monotherapy or bitherapy remains uncontrolled. Our study evaluated the efficacy of triple therapy with perindopril, amlodipine, and indapamide sustained release (SR) in patients with uncontrolled hypertension on previous antihypertensive therapy. METHODS: This 4-month, multicenter, prospective, observational, open-label study included patients switched from previous antihypertensive therapy to triple therapy with perindopril, amlodipine, and indapamide SR. The main outcome was change in office BP from baseline to 4 months, as well as changes in 24-h ambulatory BP monitoring (ABPM) parameters in a subgroup of patients. RESULTS: Age was 62.8 ± 11.3 years in 6,088 patients (55 % were male). Office BP at baseline was 158.1 ± 13.0/92.6 ± 8.8 mmHg. By 4 months, office BP decreased by 26.7 ± 13.3/12.9 ± 9.4 mmHg (p < 0.001). ABPM was performed in 62 patients. In these patients, 24-h systolic BP decreased (from 138.7 ± 12.5 to 125.5 ± 12.8 mmHg), as did 24-h diastolic BP (from 77.5 ± 11.4 to 70.4 ± 8.7 mmHg) (both p < 0.0001). Heart rate remained unchanged. In patients previously on renin-angiotensin-aldosterone system (RAAS) inhibitor/amlodipine, 24-h ambulatory systolic and diastolic BP decreased from 136.9 ± 12.8 to 125.4 ± 13.3 mmHg (p = 0.0003) and from 76.3 ± 12.6 to 70.2 ± 9.5 mmHg (p = 0.0005). In those previously on RAAS inhibitor/hydrochlorothiazide, 24-h ambulatory systolic and diastolic BP decreased from 137.8 ± 12.7 to 122.7 ± 15.4 mmHg (p = 0.0039) and from 73.6 ± 9.4 to 65.7 ± 7.3 mmHg (p = 0.002). Most (74 and 80 %, respectively) patients reached target ABPM values (<130/80 mmHg). CONCLUSION: A triple combination of perindopril, amlodipine, and indapamide SR controlled BP effectively in hypertensive patients uncontrolled by previous antihypertensive monotherapy or bitherapy, including RAAS inhibitor/amlodipine or RAAS inhibitor/hydrochlorothiazide combinations.

NADPH oxidase 4 deficiency reduces aquaporin-2 mRNA expression in cultured renal collecting duct principal cells via increased PDE3 and PDE4 activity.

The final control of renal water reabsorption occurs in the collecting duct (CD) and relies on regulated expression of aquaporin-2 (AQP2) in principal CD cells. AQP2 transcription is primarily induced by type 2 vasopressin receptor (V2R)-cAMP-protein kinase A (PKA) signaling but also by other factors, including TonEBP and NF-κB. NAPDH oxidase 4 (NOX4) represents a major source of reactive oxygen species (ROS) in the kidney. Because NOX-derived ROS may alter PKA, TonEBP and NF-κB activity, we examined the effects of NOX4 depletion on AQP2 expression. Depleted NOX4 expression by siRNA (siNOX4) in mpkCCDcl4 cells attenuated increased AQP2 mRNA expression by arginine vasopressin (AVP) but not by hypertonicity, which induces both TonEBP and NF-κB activity. AVP-induced AQP2 expression was similarly decreased by the flavoprotein inhibitor diphenyleneiodonium. siNOX4 altered neither TonEBP nor NF-κB activity but attenuated AVP-inducible cellular cAMP concentration, PKA activity and CREB phosphorylation as well as AQP2 mRNA expression induced by forskolin, a potent activator of adenylate cyclase. The repressive effect of siNOX4 on AVP-induced AQP2 mRNA expression was abolished by the non-selective phosphodiesterase (PDE) inhibitor 3-isobutyl-1-methylxanthine (IBMX) and was significantly decreased by selective PDE antagonists cilostamide and rolipram, but not vinpocetine, which respectively target PDE3, PDE4 and PDE1. Thus, by inhibiting PDE3 and PDE4 activity NOX4-derived ROS may contribute to V2R-cAMP-PKA signaling and enhance AQP2 transcription.

Complex analysis of multiple single nucleotide polymorphisms as putative risk factors of tooth agenesis in the Hungarian population.

OBJECTIVES: The role was studied of multiple single nucleotide polymorphisms in tooth agenesis in the Hungarian population using a complex approach. METHODS: Eight SNPs, PAX9 -912 C/T, PAX9 -1031 A/G, MSX1 3755 A/G, FGFR1 T/C rs881301, IRF6 T/C rs764093, AXIN2-8150 A/G, AXIN2-8434 A/G and AXIN2-30224 C/T, were studied in 192 hypodontia and 17 oligodontia cases and in 260 healthy volunteers. Case-control analysis was performed to test both allelic and genotypic associations as well as associations at the level of haplotypes. Multivariate exploratory Bayesian network-based multi-level analysis of relevance (BN-BMLA) as well as logistic regression analysis were performed. RESULTS: Conventional statistics showed that PAX9 SNP -912 C/T and the MSX1 SNP changed the incidence of hypodontia, although after Bonferroni correction for multiple hypothesis testing, the effects were only borderline tendencies. Using a statistical analysis better suited for handling multiple hypotheses, the BN-BMLA, PAX9 SNPs clearly showed a synergistic effect. This was confirmed by other multivariate analyses and it remained significant after corrections for multiple hypothesis testing (p < 0.0025). The PAX9-1031-A-PAX9-912-T haplotype was the most relevant combination causing hypodontia. Interaction was weaker between PAX9 and MSX1, while other SNPs had no joint effect on hypodontia. CONCLUSION: This complex analysis shows the important role of PAX9 and MSX1 SNPs and of their interactions in tooth agenesis, while IRF6, FGFR1 and AXIN2 SNPs had no detectable role in the Hungarian population. These results also reveal that risk factors in hypodontia need to be identified in various populations, since there is considerable variability among them.

STAT6 promotes bi-directional modulation of PKM2 in liver and adipose inflammatory cells in rosiglitazone-treated mice.

STAT6 interacts with PPARγ to elicit macrophage polarization towards an anti-inflammatory, insulin-sensitizing phenotype. Mice deficient in STAT6 display liver lipid accumulation (hepatosteatosis). Rosiglitazone (RSG), a PPARγ agonist, ameliorates hepatosteatosis and enhances insulin sensitivity. To elucidate the role of STAT6 in PPARγ action on hepatosteatosis we compared liver proteomes of RSG-treated wild type and STAT6-deficient mice and we identified pyruvate kinase M2 (PKM2), a glycolysis and proliferation-regulating enzyme that displayed STAT6-dependent expression. RSG induced PKM2 within inflammatory cells in liver but suppressed its expression in adipose tissue. RSG diminished hepatosteatosis and oxidative stress, enhanced fat accumulation and improved insulin sensitivity in STAT6-deficient mice. Our data reveal a complex interaction between STAT6 and PPARγ in the regulation of liver and adipose tissue lipid depot distribution and design STAT6 as a novel link between inflammatory cell metabolism and adipocyte and hepatocyte function.

NADPH-oxidase 4 protects against kidney fibrosis during chronic renal injury.

NADPH oxidases synthesize reactive oxygen species that may participate in fibrosis progression. NOX4 and NOX2 are NADPH oxidases expressed in the kidneys, with the former being the major renal isoform, but their contribution to renal disease is not well understood. Here, we used the unilateral urinary obstruction model of chronic renal injury to decipher the role of these enzymes using wild-type, NOX4-, NOX2-, and NOX4/NOX2-deficient mice. Compared with wild-type mice, NOX4-deficient mice exhibited more interstitial fibrosis and tubular apoptosis after obstruction, with lower interstitial capillary density and reduced expression of hypoxia-inducible factor-1α and vascular endothelial growth factor in obstructed kidneys. Furthermore, NOX4-deficient kidneys exhibited increased oxidative stress. With NOX4 deficiency, renal expression of other NOX isoforms was not altered but NRF2 protein expression was reduced under both basal and obstructed conditions. Concomitant deficiency of NOX2 did not modify the phenotype exhibited by NOX4-deficient mice after obstruction. NOX4 silencing in a mouse collecting duct (mCCD(cl1)) cell line increased TGF-ß1-induced apoptosis and decreased NRF2 protein along with expression of its target genes. In addition, NOX4 silencing decreased hypoxia-inducible factor-1α and expression of its target genes in response to hypoxia. In summary, these results demonstrate that the absence of NOX4 promotes kidney fibrosis, independent of NOX2, through enhanced tubular cell apoptosis, decreased microvascularization, and enhanced oxidative stress. Thus, NOX4 is crucial for the survival of kidney tubular cells under injurious conditions.

Comparison of three detergent-free protein extraction protocols for white adipose tissue.

A comparative study of three detergent-free protein extraction protocols--a differential centrifugal fractionation, a delipidation protocol based on the Bligh and Dyer method, and the trifluoroethanol addition as cosolvent to an aqueous buffer--was performed on white adipose tissue. The performance of the protocols directly compatible with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was evaluated based on the total protein extraction yield and the protein recovery from different functional and cellular compartments. The most suitable method for the extraction of white adipose tissue proteins from a wide range of cellular and structural compartments was the delipidation protocol based on the Bligh and Dyer method.

Impaired oral health-related quality of life in Hungary.

OBJECTIVES: To describe the prevalence of impaired oral health-related quality of life (OHRQoL) in the adult Hungarian population and to determine population-based norms for three Hungarian versions of the Oral Health Impact Profile (OHIP-H). MATERIAL AND METHODS: In a survey of 1059 randomly selected subjects, OHIP responses, age, gender, and denture status [natural dentition or fixed partial dentures (FPDs), removable partial dentures (RPDs), or complete dentures (CDs)] were collected. Study outcomes were item prevalence and OHIP summary score frequency ('norms'). RESULTS: The prevalence of OHIP items ranged from 2% to 43%, with a mean item prevalence of 14%. The distribution of summary scores was characterized by decile norms. Median scores for OHIP-H49, the 49-item questionnaire, were 6 OHIP units for subjects with FPDs, 10 OHIP units for subjects with RPDs, and 6 OHIP units for subjects with CDs. The median scores were 0, 0, and 1 OHIP units for the 14-item OHIP-H, and 0, 0, and 0 OHIP units for the five-item OHIP-H. CONCLUSIONS: Our sample demonstrates substantial OHRQoL impact in the Hungarian general population. The derived norms provide a framework for interpretation of data in future studies using the Hungarian OHIP versions, as well as data on how oral conditions and cultural factors affect perceived oral health when compared with international findings.

Differential proteomic analysis of STAT6 knockout mice reveals new regulatory function in liver lipid homeostasis.

Increased inflammatory signaling is a key feature of metabolic disorders. In this context, the role of increased pro-inflammatory signals has been extensively studied. By contrast, no efforts have been dedicated to study the contrasting scenario: the attenuation of anti-inflammatory signals and their role in metabolic homeostasis. IL-4 and IL-13 are anti-inflammatory cytokines signaling through the Signal Transducer and Activator of Transcription 6 (STAT6). Our study was aimed at evaluating the lack of STAT6 signaling on liver homeostasis. To this end we analyzed the liver proteome of wild type and STAT6 knock-out mice using 2D nanoscale LC-MS/MS with iTRAQ labeling technique. The coordinated changes in proteins identified by this quantitative proteome analysis indicated disturbed lipid homeostasis and a state of hepatocellular stress. Most significantly, the expression of the liver fatty acid binding protein (FABP1) was increased in the knock-out mice. In line with the elevated FABP1 expression we found latent liver lipid accumulation in the STAT6-deficient mice which was further aggravated when mice were challenged by a high fat diet. In conclusion, our study revealed a so far uncharacterized role for STAT6 in regulating liver lipid homeostasis and demonstrates the importance of anti-inflammatory signaling in the defense against the development of liver steatosis.

The central melanocortin system directly controls peripheral lipid metabolism.

Disruptions of the melanocortin signaling system have been linked to obesity. We investigated a possible role of the central nervous melanocortin system (CNS-Mcr) in the control of adiposity through effects on nutrient partitioning and cellular lipid metabolism independent of nutrient intake. We report that pharmacological inhibition of melanocortin receptors (Mcr) in rats and genetic disruption of Mc4r in mice directly and potently promoted lipid uptake, triglyceride synthesis, and fat accumulation in white adipose tissue (WAT), while increased CNS-Mcr signaling triggered lipid mobilization. These effects were independent of food intake and preceded changes in adiposity. In addition, decreased CNS-Mcr signaling promoted increased insulin sensitivity and glucose uptake in WAT while decreasing glucose utilization in muscle and brown adipose tissue. Such CNS control of peripheral nutrient partitioning depended on sympathetic nervous system function and was enhanced by synergistic effects on liver triglyceride synthesis. Our findings offer an explanation for enhanced adiposity resulting from decreased melanocortin signaling, even in the absence of hyperphagia, and are consistent with feeding-independent changes in substrate utilization as reflected by respiratory quotient, which is increased with chronic Mcr blockade in rodents and in humans with loss-of-function mutations in MC4R. We also reveal molecular underpinnings for direct control of the CNS-Mcr over lipid metabolism. These results suggest ways to design more efficient pharmacological methods for controlling adiposity.

Reduced expression of the NADPH oxidase NOX4 is a hallmark of adipocyte differentiation.

Adipocyte differentiation is a complex process regulated among other factors by insulin and the production of reactive oxygen species (ROS). NOX4 is a ROS generating NADPH oxidase enzyme mediating insulin's action in 3T3L1 adipocytes. In the present paper we show that NOX4 is expressed at high levels both in white and brown preadipocytes and that differentiation into adipocytes results in a decrease in their NOX4 mRNA content. These in vitro results were confirmed in vivo by demonstrating that in intact adipose tissue the majority of NOX4 expressing cells are localized within the preadipocyte containing stromal/vascular fraction, rather than in the portion consisting of mature adipocytes. In line with these observations, quantification of NOX4 mRNA in fat derived from different rodent models of insulin resistance indicated that alteration in NOX4 expression reflects changes in the ratio of adipocyte/interstitial fractions. In conclusion, we reveal that decreased NOX4 mRNA content is a hallmark of adipocyte differentiation and that NOX4 expression measured in whole adipose tissue is not an unequivocal indicator of intact or impaired insulin action.