Hepatic ChREBP orchestrates intrahepatic carbohydrate metabolism to limit hepatic glucose 6-phosphate and glycogen accumulation in a mouse model for acute Glycogen Storage Disease type Ib.

OBJECTIVE: Carbohydrate Response Element Binding Protein (ChREBP) is a glucose 6-phosphate (G6P)-sensitive transcription factor that acts as a metabolic switch to maintain intracellular glucose and phosphate homeostasis. Hepatic ChREBP is well-known for its regulatory role in glycolysis, the pentose phosphate pathway, and de novo lipogenesis. The physiological role of ChREBP in hepatic glycogen metabolism and blood glucose regulation has not been assessed in detail, and ChREBP's contribution to carbohydrate flux adaptations in hepatic Glycogen Storage Disease type 1 (GSD I) requires further investigation. METHODS: The current study aimed to investigate the role of ChREBP as a regulator of glycogen metabolism in response to hepatic G6P accumulation, using a model for acute hepatic GSD type Ib. The immediate biochemical and regulatory responses to hepatic G6P accumulation were evaluated upon G6P transporter inhibition by the chlorogenic acid S4048 in mice that were either treated with a short hairpin RNA (shRNA) directed against ChREBP (shChREBP) or a scrambled shRNA (shSCR). Complementary stable isotope experiments were performed to quantify hepatic carbohydrate fluxes in vivo. RESULTS: ShChREBP treatment normalized the S4048-mediated induction of hepatic ChREBP target genes to levels observed in vehicle- and shSCR-treated controls. In parallel, hepatic shChREBP treatment in S4048-infused mice resulted in a more pronounced accumulation of hepatic glycogen and further reduction of blood glucose levels compared to shSCR treatment. Hepatic ChREBP knockdown modestly increased glucokinase (GCK) flux in S4048-treated mice while it enhanced UDP-glucose turnover as well as glycogen synthase and phosphorylase fluxes. Hepatic GCK mRNA and protein levels were induced by shChREBP treatment in both vehicle- and S4048-treated mice, while glycogen synthase 2 (GYS2) and glycogen phosphorylase (PYGL) mRNA and protein levels were reduced. Finally, knockdown of hepatic ChREBP expression reduced starch domain binding protein 1 (STBD1) mRNA and protein levels while it inhibited acid alpha-glucosidase (GAA) activity, suggesting reduced capacity for lysosomal glycogen breakdown. CONCLUSIONS: Our data show that ChREBP activation controls hepatic glycogen and blood glucose levels in acute hepatic GSD Ib through concomitant regulation of glucose phosphorylation, glycogenesis, and glycogenolysis. ChREBP-mediated control of GCK enzyme levels aligns with corresponding adaptations in GCK flux. In contrast, ChREBP activation in response to acute hepatic GSD Ib exerts opposite effects on GYS2/PYGL enzyme levels and their corresponding fluxes, indicating that GYS2/PYGL expression levels are not limiting to their respective fluxes under these conditions.

Influence of a downstream narrowing on the flow profile in a tube.

The distance over which the upstream flow conditions in a tube are disturbed by a stenosis downstream, i.e. the outlet length, was investigated for Reynolds numbers in the range 210-2900. Two methods were used, the Navier-Stokes equations were solved with a computer and a physical model was constructed and maximal velocities were measured with an ultrasound Doppler system. The computer model showed that Re number does not influence the outlet length, varying the stenosis area from 25% to 90% has an effect. However, the outlet length remained small, below 70% of the diameter of the tube. The physical model confirmed for a 75% stenosis that the outlet length is small, this method set the limit at not more than 1.2 times the tube diameter.

Design and wear testing of a temporomandibular joint prosthesis articulation.

As part of the development of a total temporomandibular joint prosthesis, a prosthesis articulation was designed. The articulation consists of a spherical head (ball) of the mandibular part, rotating against an enveloping ultra-high-molecular-weight polyethylene (UHMWPE) disc with a flat cranial side, which slides along the opposing skull part. The aim of this study was to determine the in vitro wear rate of the articulation, and to predict the in vivo wear rate from the results. Based on a disc thickness of 5 mm and a ball diameter of 8 mm, the stresses within the disc were calculated by means of a finite element computer model. The wear rate of the ball-disc articulation was determined by in vitro wear tests, with a stainless-steel ball rotating against a UHMWPE disc in a serum-based lubricant. Eight discs were tested for seven million cycles each. The in vitro wear rate of the disc-skull part articulation was calculated from the test results of the ball-disc articulation. The maximum Von Mises' stress was less than the yield strength of UHMWPE and, therefore, was sufficiently low. The in vitro wear rate of the ball-disc articulation was 0.47 mm3 per million cycles. The in vivo expected total wear rate is 0.65 mm3 per year, corresponding with a yearly decrease of disc thickness of 0.0094 mm. Although it is difficult to judge whether this wear rate is sufficiently low, because the influence of UHMWPE wear particles in the TMJ region is unknown, both the expected wear rate and the decrease of thickness appear to be acceptable.

Analysis of the mechanical behavior of the Nijdam voice prosthesis.

The valveless Nijdam prosthesis is a new voice prosthesis for laryngectomized patients using tracheoesophageal speech. An "umbrella-like hat" covers the esophageal side of the tracheoesophageal fistula and is deformed during speech by air pressure. To decrease pressure loss during speech, a good understanding of the mechanical behavior is essential. In the present study, the Finite Element Method (FEM), used in engineering to analyze the mechanical behavior of complex structures, was applied to analyze eight possible improvements of the Nijdam prosthesis. This study found that, during speech, deformation of hat and soft tissue occur. Distinct differences in the hat's deformation of the eight models also were found. It is concluded that complex structures like the Nijdam prosthesis can be analyzed by FEM. An optimal model was found to decrease pressure loss while stresses in the device remain safe.

Alternative voice after laryngectomy using a sound-producing voice prosthesis.

OBJECTIVE: To improve the voice quality of female laryngectomees and/or laryngectomees with a hypotonic pharyngoesophageal (PE) segment by means of a pneumatic artificial source of voice incorporated in a regular tracheoesophageal (TE) shunt valve. STUDY DESIGN: Experimental, randomized, crossover trial. METHODS: The new sound source consists of a single silicone lip, which performs an oscillatory movement driven by expired pulmonary air flowing along the outward-striking lip through the TE shunt valve. A prototype of this pneumatic sound source is evaluated in vitro and in six laryngectomees. In vivo evaluation includes speech rate, maximal phonation time, perceptual voice evaluation of read-aloud prose by an expert listener, speech intelligibility measurements with 12 listeners, and self-assessment by the patients. Moreover, extensive acoustical and aerodynamic in vivo registrations are performed using a newly developed data acquisition system. RESULTS: The current prototype seems beneficial in female laryngectomees with a hypotonic PE segment only. For them the sound-producing voice prosthesis improves voice quality and increases the average pitch of voice, without decreasing intelligibility or necessitating other pressure and airflow rates than regular TE shunt speech. Pitch regulation of this prosthetic voice is possible, yet limited. CONCLUSIONS: The mechanism is feasible and does not result in unacceptable airflow resistance. For this new mechanism of alaryngeal voice to become an established technique for postlaryngectomy voice restoration, a voice suitably pitched for male laryngectomees has to be generated and a large part of the melodic and dynamic range of the sound source has to be attainable within physiological airflow rates.

Design and in vitro testing of a voice-producing element for laryngectomized patients.

A voice-producing element has been developed to improve speech quality after laryngectomy. The design process started with the formulation of a list of requirements. The lip principle has the best potential for fulfilling the requirements. A numerical model was made to find the optimal geometry of an element based on the lip principle. Extensive in vitro tests were performed to check all requirements. For this a test set-up with realistic acoustic and aerodynamic properties was developed. Results show that the protruding lip length dominates fundamental frequency, cross-sectional area dominates flow resistance and relation between flow and fundamental frequency. Most requirements have been fulfilled; both for males and females a potentially good functioning prototype could be selected. Clinical experiments will be performed to confirm the quality of the voice-producing prosthesis.

Resistance of quiescent and proliferating airway epithelial cells to H2O2 challenge.

Alveolar epithelial cell injury and recovery are important in the pathogenesis of oxidant-induced lung damage. The alveolar cell line A549 was used to study responses of proliferating and quiescent cells in culture to time- and dose-dependent hydrogen peroxide (H(2)O(2)) challenges. Recovery was monitored after 24 h of incubation in fresh medium with 10% serum. The adherent cells were counted and the resistance and recovery of the attached cells was assessed by appearance, by measuring the number of viable, apoptotic and necrotic cells using fluorescent-activated cell sorting, and by determining the intracellular free thiol content. A549 cells recovered from a 1-h challenge with up to 1 mM H(2)O(2) but could not sustain a more prolonged challenge (6 or 24 h) with 0.5 mM or 1.0 mM H(2)O(2). These more severe conditions resulted in: loss of cells by detachment from the plate surface; reduced numbers of viable cells primarily due to necrosis; and a strong reduction of the intracellular free thiol content. Quiescent cells proved to be more sensitive to oxidative stress than proliferating cells. Intracellular free thiol levels apparently play a decisive role in cell survival, preferentially protecting proliferating cells.

The prevalence of house dust mite (HDM) allergy and the use of HDM-impermeable bed covers in a primary care population of patients with persistent asthma in the Netherlands.

BACKGROUND: : House dust mite (HDM) allergen is one of the most common allergens to which asthma patients are sensitised. Prevalence of HDM allergy varies in the literature. Use of HDM-impermeable bed covers reduces exposure to HDM allergen. The aim of this study was to assess the prevalence of HDM allergy in a primary care population of asthma patients, as well as the use of HDM-impermeable bed covers by these patients. METHODS: : A random sample of asthma patients between 16 and 60 years old was taken from general practices. Allergy was assessed with a radio allegro sorbent test (RAST). A questionnaire was used to identify demographic characteristics and the actual use of bed covers. RESULTS: : 534 patients were invited and 160 patients participated. 53 patients not willing to participate were randomly selected to test the external validity of our findings. The sample was representative for the primary care asthma population. 48.8% of the asthma population was sensitised to HDM allergen. 25.6% of the HDM-allergic asthma patients were using HDM-impermeable bed covers. CONCLUSION: : Almost half of the patients with asthma were sensitised to HDM allergen. Only a minority of the patients used HDM-impermeable bed covers.

Glottal flow through a two-mass model: comparison of Navier-Stokes solutions with simplified models.

A new numerical model of the vocal folds is presented based on the well-known two-mass models of the vocal folds. The two-mass model is coupled to a model of glottal airflow based on the incompressible Navier-Stokes equations. Glottal waves are produced using different initial glottal gaps and different subglottal pressures. Fundamental frequency, glottal peak flow, and closed phase of the glottal waves have been compared with values known from the literature. The phonation threshold pressure was determined for different initial glottal gaps. The phonation threshold pressure obtained using the flow model with Navier-Stokes equations corresponds better to values determined in normal phonation than the phonation threshold pressure obtained using the flow model based on the Bernoulli equation. Using the Navier-Stokes equations, an increase of the subglottal pressure causes the fundamental frequency and the glottal peak flow to increase, whereas the fundamental frequency in the Bernoulli-based model does not change with increasing pressure.

Does food intolerance play a role in juvenile chronic arthritis?

Sixty children with juvenile chronic arthritis (JCA) have been examined at the paediatric rheumatology out-patient clinic in Maastricht, of whom three ultimately appeared to have a food intolerance. In one of these three patients, there appeared to be a relationship with joint complaints. In the course of the elimination/challenge tests which were conducted, severe painful swelling of the knee occurred rapidly after each challenge. Three challenges were carried out with the same result each time. Since the symptoms did not disappear entirely following elimination of milk, it was concluded that milk intolerance in this case was an aggravating factor in a seronegative monoarticular JCA. In the second and third patients, a strict diet had no positive effect on the joint problems. In conclusion, the existence of such a connection between food and chronic joint complaints has been made clear, it only plays a role in incidental cases.

In vitro measurements of aerodynamic characteristics of an improved tracheostoma valve for laryngectomees.

Tracheostoma valves are often required in the rehabilitation process of speech after total laryngectomy. Patients are thus able to speak without using their hands to close the tracheostoma. The improved Groningen tracheostoma valve consists of a "cough" valve with an integrated ("speech") valve, which closes for phonation. The cough valve opens as the result of pressure produced by the lungs during a cough. The speech valve closes by the airflow produced by the lungs, thus directing air from the lungs into the esophagus at a deliberately chosen moment. An experimental setup with a computer-based acquisition program was developed to measure the pressure at which the cough valve opened and the flow at which the speech valve closed. In addition, the airflow resistance coefficient of the tracheostoma valve was defined and measured with an open speech valve. Both dry air from a cylinder and humid expired air were used. Results showed a pressure range of 1-7 kPa to open the cough valve and a flow range of 1.2-2.7 l/s to close the speech valve. These values were readily attained during speech, while the flow range occurred above values reached in quiet breathing. The device appeared to function well in physiological ranges and was optimally adjustable to an individual setting. No significant differences were measured between air from a cylinder and humid expired air. Findings showed that methods used to obtain results could be employed as a reference method for comparing aerodynamic characteristics of tracheostoma valves.

Female-pitched sound-producing voice prostheses--initial experimental and clinical results.

In order to improve voice quality in female laryngectomees and/or laryngectomees with a hypotonic pharyngo-oesophageal segment, a sound-producing voice prosthesis was designed. The new source of voice consists of either one or two bent silicone lips which perform an oscillatory movement driven by the expired pulmonary air that flows along the outward-striking lips through the tracheo-oesophageal shunt valve. Four different prototypes of this pneumatic sound source were evaluated in vitro and in two female laryngectomees, testing the feasibility and characteristics of this new mechanism for alternative alaryngeal voice production. In vivo evaluation included acoustic analyses of both sustained vowels and read-aloud prose, videofluoroscopy, speech rate, and registration of tracheal phonatory pressure and vocal intensity. The mechanism proved feasible and did not result in unacceptable airflow resistance. The average pitch of voice increased and clarity improved in female laryngectomees. Pitch regulation of this prosthetic voice is possible with sufficient modulation to avoid monotony. The quality of voice attained through the sound-producing voice prostheses depends on a patient's ability to let pulmonary air flow easily through the pharyngo-oesophageal segment without evoking the low-frequency mucosal vibrations that form the regular tracheo-oesophageal shunt voice. These initial experimental and clinical results provide directions for the future development of sound-producing voice prostheses. A single relatively long lip in a container with a rectangular lumen that hardly protrudes from the voice prosthesis may have the most promising characteristics.