The influence of mouth breathing on skeletal and dental features of splanchnocranium.

OBJECTIVE: This study was performed to investigate changes in dentofacial characteristics associated with mouth breathing (MB) and adenoidectomy. BACKGROUND: MB is considered to be an etiological factor of malocclusion. Adenoidectomy is supposed to have the ability to prevent the development of dentofacial deformities. METHODS: This retrospective study included 123 patients, namely 57 nose breathers, 19 former mouth breathers, who have undergone adenoidectomy, and 47 mouth breathers. The groups were compared according to their skeletal and dental characteristics. The measurements of each individual were obtained from lateral cephalograms and dental casts. The comparison was done using one-way ANOVA, Bonferroni post-hoc, Mann-Whitney U and Kruskal-Wallis tests. The statistically significant difference was defined as p<0.05. RESULTS: The MB group showed an increase in ArGoMe (p=0.02) angle. No difference was found in the sagittal parameters among the groups. Upper dental arch compression was positively correlated with MB(p=0.00), even in adenoidectomy cases (p=0.01). CONCLUSION: MB alters the vertical and transverse growth of the craniofacial complex. It is associated with longer lower anterior facial height and decreased maxillary intermolar distance. However, it does not influence the sagittal parameters. Airway clearance via adenoidectomy promotes the normalization of vertical parameters (Tab. 1, Fig. 2, Ref. 20).

Do we have to reduce the recall period? Validity of a daily physical activity questionnaire (PAQ24) in young active adults.

BACKGROUND: Combining the strengths of physical activity (PA) diaries and questionnaires may be needed to improve the unsatisfying measurement quality of existing PA questionnaires. This study investigated the construct validity of a short PA questionnaire (Physical Activity Questionnaire for 24 h [PAQ24]) with a recall period of one day. METHODS: In this cross-sectional study, participants completed the PAQ24 on seven consecutive days while wearing an accelerometer (GENEActiv). Thereafter, the Global Physical Activity Questionnaire (GPAQ) was completed. Spearman correlation coefficients and Bland-Altman analysis were used to assess construct validity. RESULTS: Overall, 50 active adults (11 women, mean age = 25.1 ± 2.5) participated. Relative agreements between Total PA of PAQ24 and accelerometer were 0.37 ≤ ρ ≤ 0.72 for each day with satisfying agreement on five out of seven days. Weekly relative agreement for Total PA was moderate (ρ = 0.44). Relative agreements between PAQ24 and GPAQ were ρ = 0.43 for Total PA. Daily and weekly absolute agreements were poor indicated by wide limits of agreement. CONCLUSIONS: In contrast to weekly Total PA, the majority of daily results of the PAQ24 showed satisfying construct validity. A short recall period may improve the measurement quality of PA questionnaires, but measurement errors and the costs of multiple administrations must be considered in future studies.

Somatic mutations in kinetochore gene KNSTRN are associated with basal proliferating actinic keratoses and cutaneous squamous cell carcinoma.

BACKGROUND: Mutations in kinetochore gene KNSTRN accelerate the development of cutaneous squamous cell carcinoma (SCC) and may correlate with different histological classifications of actinic keratosis (AKs). OBJECTIVE: To determine KNSTRN gene mutation frequency in healthy skin (HS), actinically damaged skin (ADS), in AKs with different histomorphological gradings and invasive SCCs. METHODS: All samples were histologically evaluated. AK lesions were additionally classified according to their upwards (AK I-III) and downwards (PRO I-III) directed growth pattern. Mutation analyses of all samples were performed using the Sanger method. RESULTS: With one exception, all detected mutations in KNSTRN gene showed an alanine-to-glutamate substitution at codon 40 (p.Ala40Glu). p.Ala40Glu mutation was found in 6.9% (2/29) of HS, in 16.1% (5/31) of ADS, in 18.3% (20/109) of AKs and in 30.0% (9/30) of invasive SCCs. Further stratification of AKs using the common AK classification of Röwert-Huber revealed the p.Ala40Glu mutation in 14.7% (5/43), 13.3% (4/30) and 24.4% (11/45) (AK I, II and III). In contrast, the new PRO classification showed a distribution of 3.6% (1/28) in PRO I, 21.7% (13/60) in PRO II and 28.6% (6/21) in PRO III. Mutation frequency in HS showed significant differences compared to AKs classified as PRO III and invasive SCCs (P < 0.05). In contrast, there were no statistically significant differences between HS and AKs when classified according to Röwert-Huber. CONCLUSIONS: Recurrent somatic mutation p.Ala40Glu in KNSTRN gene is associated with basal proliferating AKs in accordance with invasive SCCs. This supports the impact of basal proliferative pattern in terms of progression.

The influence of different illumination parameters on protoporphyrin IX induced cell death in squamous cell carcinoma cells.

BACKGROUND: Topical photodynamic therapy (PDT) is a highly effective therapy especially for extended cancerized fields of the skin. Whenever extended fields are treated pain management is advisable. Light source mediated pain management can be performed by reducing fluence rates, as long as this does not compromise efficacy. METHODS: Two squamous cell carcinoma cell lines (A431 and SCC-13) were subjected to in vitro PDT using two different ALA concentrations and synthesis intervals and protoporphyrin IX (PpIX) synthesis was assessed. Two total light doses (6 J/cm2 and 37 J/cm2) were applied at three different fluence rates and cell viability was measured using the MTS-test. RESULTS: Both cell lines synthetized PpIX at different kinetics. A431 cells produced a maximum 28.6 nmol/l PpIX, while SCC-13 reached only a production of 8.7 nmol/l. Illumination reduced cell viability depending on PpIX content and light dose. When a lower light dose (6 J/cm2) was applied, only the combination with the highest PpIX content was effective in A431 cells and no effect could be detected in SCC-13 cells. With a light dose of 37 J/cm2, lower PpIX amounts became effective in A431 and cell death could be induced in SCC-13 cells. Light fluence rate had no differential effect in this setup. CONCLUSIONS: In both, A431 and SCC-13 cells, total light dose is a key factor for photodynamic efficacy. Additionally, our results hint towards a threshold concentration of PpIX upon which a drastic loss of viability occurs. Light fluence rate in the analyzed range is not a limiting factor of photodynamic cytotoxicity. This may allow for the clinical implementation of low fluence rate protocols for pain management without compromising efficacy.

Patient-reported health outcomes in patients with non-melanoma skin cancer and actinic keratosis: results from a large-scale observational study analysing effects of diagnoses and disease progression.

BACKGROUND: Non-melanoma skin cancer (NMSC) and actinic keratosis (AK) are very common among fair-skinned individuals. A disease continuum from AK to squamous cell carcinoma (SCC) has been frequently postulated. AK and NMSC may influence quality of life (QL) of patients, and it can be suspected that disease progression entails a QL reduction. The purpose of this study was to document QL in patients with NMSC and AK using the health-outcome questionnaire EQ-5D-5L. METHODS: The study was designed as a non-interventional, prospective, cross-sectional study. Patients with AK, SCC, basal cell carcinoma (BCC) or multiple diagnoses were enrolled in this study in 29 dermatological centres across Germany. Patients were asked to complete the EQ-5D-5L (compromising EQ Index and EQ VAS), and the dermatologists provided diagnosis, disease history and treatment data. RESULTS: A total of 1184 patients were enrolled and diagnosed as follows: 73% AK, 49% BCC and 17% SCC. 66% had a single diagnosis, 28% two different diagnoses and 6% three different diagnoses. QL was strongly associated with patients' diagnosis. Patients with a single AK diagnosis had significantly higher mean EQ VAS (78) than patients with BCC (74), SCC (72), and BCC plus SCC (69), P < 0.050. When the effects of disease progression were calculated, patients with AK plus SCC reported significantly less mean EQ VAS (71) than patients with a single AK diagnosis (78), P < 0.011. CONCLUSIONS: While rarely being imminently life-threatening, NMSC and AK have an impact on QL as quantified by the EQ-5D-5L. This impact is associated with diagnosis (AK vs. NMSC) and clinical progression (AK vs. AK plus SCC). Both lead to a clear decline in QL. This shows that disease progression is perceived and judged as detrimental by patients and that AK and NMSC should be diligently treated to preserve and restore QL.

A phase I/II study of KW-2478, an Hsp90 inhibitor, in combination with bortezomib in patients with relapsed/refractory multiple myeloma.

BACKGROUND: KW-2478 is a novel non-ansamycin Hsp90 inhibitor with modest single-agent activity in relapsed/refractory myeloma but which shows synergistic antimyeloma activity with bortezomib (BTZ) in preclinical studies. This study determined the safety, preliminary clinical activity, and pharmacokinetics of KW-2478, an Hsp90 inhibitor, in combination with BTZ in patients with relapsed/refractory multiple myeloma (MM). METHODS: Phase I dose escalation determined the recommended phase II dose (RP2D) of KW-2478 plus BTZ, which was then used during phase II. RESULTS: The maximum tolerated dose was not reached during phase I and the RP2D was KW-2478 175 mg m-2 plus BTZ 1.3 mg m-2 on days 1, 4, 8, and 11 every 3 weeks. In the efficacy evaluable phase I/II population treated at the RP2D (n=79), the objective response rate was 39.2% (95% confidence interval: 28.4-50.9%), clinical benefit rate 51.9% (40.4-63.3%), median progression-free survival 6.7 (5.9-not reached (NR)) months, and median duration of response 5.5 (4.9-NR) months. In the phase I/II safety population (n=95), the most frequently observed treatment-related grade 3/4 adverse events were diarrhoea, fatigue, and neutropenia (each in 7.4% of patients), and nausea and thrombocytopenia (each in 5.3%). CONCLUSIONS: KW-2478 plus BTZ was well tolerated with no apparent overlapping toxicity in patients with relapsed/refractory MM. The antimyeloma activity of KW-2478 in combination with BTZ as scheduled in this trial appeared relatively modest; however, the good tolerability of the combination would support further exploration of alternate dosing schedules and combinations.

Designing and evaluating Brain Powered Games for cognitive training and rehabilitation in at-risk African children.

BACKGROUND: Valid, reliable, accessible, and cost-effective computer-training approaches can be important components in scaling up educational support across resource-poor settings, such as sub-Saharan Africa. The goal of the current study was to develop a computer-based training platform, the Michigan State University Games for Entertainment and Learning laboratory's Brain Powered Games (BPG) package that would be suitable for use with at-risk children within a rural Ugandan context and then complete an initial field trial of that package. METHODS: After game development was completed with the use of local stimuli and sounds to match the context of the games as closely as possible to the rural Ugandan setting, an initial field study was completed with 33 children (mean age = 8.55 ± 2.29 years, range 6-12 years of age) with HIV in rural Uganda. The Test of Variables of Attention (TOVA), CogState computer battery, and the Non-Verbal Index from the Kaufman Assessment Battery for Children, 2nd edition (KABC-II) were chosen as the outcome measures for pre- and post-intervention testing. The children received approximately 45 min of BPG training several days per week for 2 months (24 sessions). RESULTS: Although some improvements in test scores were evident prior to BPG training, following training, children demonstrated clinically significant changes (significant repeated-measures outcomes with moderate to large effect sizes) on specific TOVA and CogState measures reflecting processing speed, attention, visual-motor coordination, maze learning, and problem solving. CONCLUSIONS: Results provide preliminary support for the acceptability, feasibility, and neurocognitive benefit of BPG and its utility as a model platform for computerized cognitive training in cross-cultural low-resource settings.

Comparison of the uptake of 5-aminolevulinic acid and its methyl ester in keratinocytes and skin.

Photodynamic therapy is widely used in the treatment of superficial skin cancers. 5-Aminolevulinic acid (ALA) and its methylated form, methyl-ALA (MAL), are frequently used as precursors to photosensitizing substances. Nevertheless, the mechanism of the uptake of ALA and MAL in keratinocytes and of their skin penetration is still controversial. Since both compounds are not sufficiently lipophilic to penetrate through lipid membranes, they must employ specific uptake systems which may vary between different cell types. Here, we studied ALA and MAL uptake in keratinocyte cell lines originating from healthy cells (CCD 1106 KERTr cells) or keratinocyte tumors (A431 cells). ALA uptake resulted in faster protoporphyrin IX (PpIX) production than MAL uptake. A pharmacological characterization of the uptake systems revealed that PpIX formation was most efficiently reduced with GABA transporter (GAT) substrates. GABA, ß-alanine, and (S)-SNAP-5114 reduced ALA uptake and, to a lesser extent, MAL uptake in the cell lines. The pharmacology of these compounds indicates that ALA and MAL are taken up by normal and pathological keratinocytes via GAT-3. Furthermore, the amino acids arginine, cysteine, and histidine also inhibited the uptake of ALA, and even more so MAL, suggestive of an additional involvement of amino acid transporters. To show that PpIX formation in vivo is restricted to the application site, which has been questioned for ALA in one other report, we applied clinically used ALA and MAL formulations to the skin of nude mice. Contrary to the results of these previous authors, the resulting PpIX fluorescence increased over time and was restricted to the application site for both preparations.

δ-Aminolevulinic acid and its methyl ester induce the formation of Protoporphyrin IX in cultured sensory neurones.

Application of δ-aminolevulinic acid (ALA) or its methyl ester (MAL) onto cutaneous tumours increases intracellular Protoporphyrin IX (PpIX), serving as photosensitizer in photodynamic therapy (PDT). While PDT is highly effective as treatment of neoplastic skin lesions, it may induce severe pain in some patients. Here, we investigated ALA and MAL uptake and PpIX formation in sensory neurones as potential contributor to the pain. PpIX formation was induced in cultured sensory neurones from rat dorsal root ganglion by incubation with ALA or MAL. Using inhibitors of GABA transporters (GAT), a pharmacological profile of ALA and MAL uptake was assessed. GAT mRNA expression in the cultures was determined by RT-PCR. Cultured sensory neurones synthesised Protoporphyrin IX (PpIX) from extracellularly administered ALA and MAL. PpIX formation was dose- and time-dependent with considerably different kinetics for both compounds. While partial inhibition occurred using L-arginine, PpIX formation from both ALA and MAL could be fully blocked by the GABA-Transporter (GAT)-2/3 inhibitor (S)-SNAP 5114 with similar K (i) (ALA: 195 ± 6 µM; MAL: 129 ± 13 µM). GAT-1 and GAT-3 could be detected in sensory neurons using RT-PCR on mRNA level and using [³H]-GABA uptake on protein level. Cultured sensory neurones take up ALA and MAL and synthesize PpIX from both, enabling a direct impact of photodynamic therapy on cutaneous free nerve endings. The pharmacological profile of ALA and MAL uptake in our test system was very similar and suggests uptake via GABA and amino acid transporters.

Diagnostic single nucleotide polymorphisms for identifying westslope cutthroat trout (Oncorhynchus clarki lewisi), Yellowstone cutthroat trout (Oncorhynchus clarkii bouvieri) and rainbow trout (Oncorhynchus mykiss).

We describe 12 diagnostic single nucleotide polymorphism (SNP) assays for use in species identification among rainbow and cutthroat trout: five of these loci have alleles unique to rainbow trout (Oncorhynchus mykiss), three unique to westslope cutthroat trout (O. clarkii lewisi) and four unique to Yellowstone cutthroat trout (O. clarkii bouvieri). These diagnostic assays were identified using a total of 489 individuals from 26 populations and five fish hatchery strains.

Exhaled methyl nitrate as a noninvasive marker of hyperglycemia in type 1 diabetes.

Recent technical advances allow detection of several hundred volatile organic compounds (VOCs) in human exhaled air, many of which reflect unidentified endogenous pathways. Our group has previously estimated plasma glucose levels in healthy adults during a standard oral glucose tolerance test via exhaled VOC analysis. As a result of the metabolic characteristics of hyperglycemia in the diabetic (low insulin and increased free fatty acids and ketones), we hypothesized that different exhaled VOC profiles may be present in children with type 1 diabetes mellitus (T1DM) during spontaneous hyperglycemia. Exhaled methyl nitrate strongly correlated specifically with the acute, spontaneous hyperglycemia of T1DM children. Eighteen experiments were conducted among 10 T1DM children. Plasma glucose and exhaled gases were monitored during either constant euglycemia (n = 5) or initial hyperglycemia with gradual correction (n = 13); all subjects received i.v. insulin and glucose as needed. Gas analysis was performed on 1.9-liter breath samples via gas chromatography using electron capture, flame ionization, and mass selective detection. Among the approximately 100 measured exhaled gases, the kinetic profile of exhaled methyl nitrate, commonly present in room air in the range of 5-10 parts per trillion, was most strongly statistically correlated with that of plasma glucose (P = 0.003-0.001). Indeed, the kinetic profiles of the two variables paralleled each other in 16 of 18 experiments, including repeat subjects who at different times displayed either euglycemia or hyperglycemia.

Computerized evaluation of mammographic lesions: what diagnostic role does the shape of the individual microcalcifications play compared with the geometry of the cluster?

OBJECTIVE: The objective of this study was to compare the diagnostic role of features reflecting the geometry of clusters with features reflecting the shape of the individual microcalcification in a mammographic computer-aided diagnosis system. MATERIALS AND METHODS: Three hundred twenty-four cases of clustered microcalcifications with biopsy-proven results were digitized at 42-microm resolution and analyzed on a computerized system. The shape factor and number of neighbors were computed for each microcalcification, and the eccentricity of the cluster was computed as well. The shape factor is related to the individual microcalcification; the average number of neighbors and the cluster eccentricity reflect the cluster geometry. Stepwise discriminant analysis was used to evaluate the contribution of the extracted features in predicting malignancy. The performance of a classifier based on the features selected by stepwise discriminant analysis was evaluated by receiver operating characteristic (ROC) analysis. RESULTS: To obtain the best discrimination model, we used stepwise discriminant analysis to select the average number of neighbors and the shape of the individual microcalcification, but excluded cluster eccentricity. A classification scheme assigned the average number of neighbors a weighting factor, which was 1.49 times greater than that assigned to the shape factor of the individual microcalcification. A scheme based only on these two features yielded an ROC curve with an area under the curve (A(z)) of 0.87, indicating a positive predictive value of 61% for 98% sensitivity. CONCLUSION: Computerized analysis permitted calculations reflecting the shape of individual microcalcification and the geometry of clusters of microcalcifications. For the computerized classification scheme studied, the cluster geometry was more effective in differentiating benign from malignant clusters than was the shape of individual microcalcification.

Modelling the controls of the eukaryotic cell cycle.

The eukaryotic cell-division cycle is regulated by three modules that control G(1)/S, G(2)/M and meta/anaphase transitions. By using mathematical modelling, we show the dynamic characteristics of these individual modules and we also assemble them together into a comprehensive model of the eukaryotic cell-division cycle. With this comprehensive model, we also discuss the mechanisms by which different checkpoint pathways stabilize different cell-cycle states and inhibit the transitions that drive cell-cycle progression.

Regularities and irregularities in the cell cycle of the fission yeast, Schizosaccharomyces pombe (a review).

In an exponentially growing wild-type fission yeast culture a size control mechanism ensures that mitosis is executed only if the cells have reached a critical size. However, there is some scattering both in cell length at birth (BL) and in cycle time (CT). By computational simulations we show here that this scattering cannot be explained solely by asymmetric cell division, therefore we assume that nuclear division is a stochastic, asymmetric process as well. We introduce an appropriate stochastic variable into a mathematical model and prove that this assumption is suitable to describe the CT vs. BL graph in a wild-type fission yeast population. In a double mutant of fission yeast (namely wee1-50 cdc25 delta) this CT vs. BL plot is even more curious: cycle time splits into three different values resulting in three clusters in this coordinate system. We show here that it is possible to describe these quantized cycles by choosing the appropriate values of the key parameters of mitotic entry and exit and even more the clustered behavior may be simulated by applying a further stochastic parameter.

Network dynamics and cell physiology.

Complex assemblies of interacting proteins carry out most of the interesting jobs in a cell, such as metabolism, DNA synthesis, movement and information processing. These physiological properties play out as a subtle molecular dance, choreographed by underlying regulatory networks. To understand this dance, a new breed of theoretical molecular biologists reproduces these networks in computers and in the mathematical language of dynamical systems.

A stochastic, molecular model of the fission yeast cell cycle: role of the nucleocytoplasmic ratio in cycle time regulation.

We propose a stochastic version of a recently published, deterministic model of the molecular mechanism regulating the mitotic cell cycle of fission yeast, Schizosaccharomyces pombe. Stochasticity is introduced in two ways: (i) by considering the known asymmetry of cell division, which produces daughter cells of slightly different sizes; and (ii) by assuming that the nuclear volumes of the two newborn cells may also differ. In this model, the accumulation of cyclins in the nucleus is proportional to the ratio of cytoplasmic to nuclear volumes. We have simulated the cell-cycle statistics of populations of wild-type cells and of wee1(-) mutant cells. Our results are consistent with well known experimental observations.

Angiotensin-converting enzyme gene polymorphism in patients with systemic lupus.

The renin-angiotensin-aldosterone system (RAAS) has been considered one of the probable pathophysiologic mechanisms involved in disease progression. Genetic polymorphism of the RAAS has been associated with the clinical course of renal disease. One of the genetic polymorphisms is a deletion or insertion of a 287 base pair fragment in intron 16 of the angiotensin-converting enzyme (ACE) gene. It is known that ACE gene polymorphism is present in humans and that it is associated with an increased risk of cardiovascular diseases, renal disease progression and sarcoidosis. In this study, the potential significance of ACE gene polymorphism in patients with systemic lupus erythematosus (SLE) was investigated. ACE gene polymorphism was determined in 18 patients with SLE and in 21 healthy volunteers as a control group. The mean age of patients was 38.5 years. All patients had a mean follow-up of 30.7 +/- 20.2 months (range 5-95 months). ACE genotypes were determined by the method of polymerase chain reaction. Proteinuria and creatinine were also followed. The frequency of DD, ID and II genotypes was 50%, 28% and 22% in SLE patients and 25%, 50% and 25% in healthy controls, respectively. DD genotype was more common in SLE patients than in the control group. The patients with II genotype had lower proteinuria and creatinine level than those with DD genotype (p < 0.05). The time to disease remission was shorter in patients with II genotype (p < 0.05). Study results indicated an increased frequency of D allele in SLE patients. The increased ACE activity in these patients pointed to the need of further studies of ACE gene polymorphism in SLE.

Measuring comfort in caregivers and patients during late end-of-life care.

The purpose of this study was to test several formats of end-of-life comfort instruments for patients and closely involved caregivers. Kolcaba's Comfort Theory was the theoretical framework utilized. Different response formats for two end-of-life (EOL) comfort questionnaires (for patients and caregivers, respectively), and horizontal and vertical visual analog scales for total comfort (TC) lines were compared in two phases. Evaluable data were collected from both members of 38 patient-caregiver dyads in each phase. Suitable dyads were recruited from two hospice agencies in northeastern Ohio. Cronbach's alpha for the EOL comfort questionnaire (six response Likert-type format) tested during phase I for patients was .98 and for caregivers was .97. Test-retest reliability for the vertical TC line tested during phase I for patients was .64 and for caregivers was .79. The implications of this study for nursing practice and research are derived from the American Nursing Association (ANA) position statement about EOL care, which states that comfort is the goal of nursing for this population. These instruments will be useful for assessing comfort in actively dying patients and comfort of their caregivers as well as for developing evidence-based practice for this population.

Regulation of the eukaryotic cell cycle: molecular antagonism, hysteresis, and irreversible transitions.

In recent years, molecular biologists have uncovered a wealth of information about the proteins controlling cell growth and division in eukaryotes. The regulatory system is so complex that it defies understanding by verbal arguments alone. Quantitative tools are necessary to probe reliably into the details of cell cycle control. To this end, we convert hypothetical molecular mechanisms into sets of nonlinear ordinary differential equations and use standard analytical and numerical methods to study their solutions. First, we present a simple model of the antagonistic interactions between cyclin-dependent kinases and the anaphase promoting complex, which shows how progress through the cell cycle can be thought of as irreversible transitions (Start and Finish) between two stable states (G1 and S-G2-M) of the regulatory system. Then we add new pieces to the "puzzle" until we obtain reasonable models of the control systems in yeast cells, frog eggs, and cultured mammalian cells.