Faulty neuronal determination and cell polarization are reverted by modulating HD early phenotypes.

Increasing evidence suggests that early neurodevelopmental defects in Huntington's disease (HD) patients could contribute to the later adult neurodegenerative phenotype. Here, by using HD-derived induced pluripotent stem cell lines, we report that early telencephalic induction and late neural identity are affected in cortical and striatal populations. We show that a large CAG expansion causes complete failure of the neuro-ectodermal acquisition, while cells carrying shorter CAGs repeats show gross abnormalities in neural rosette formation as well as disrupted cytoarchitecture in cortical organoids. Gene-expression analysis showed that control organoid overlapped with mature human fetal cortical areas, while HD organoids correlated with the immature ventricular zone/subventricular zone. We also report that defects in neuroectoderm and rosette formation could be rescued by molecular and pharmacological approaches leading to a recovery of striatal identity. These results show that mutant huntingtin precludes normal neuronal fate acquisition and highlights a possible connection between mutant huntingtin and abnormal neural development in HD.

In vivo delivery of DN:REST improves transcriptional changes of REST-regulated genes in HD mice.

Current therapeutic strategies for Huntington's disease (HD) are focused on symptom management of disease progression. Transcriptional dysregulation is one of the major characteristics in HD. REST is a transcriptional repressor that silences gene expression through binding to RE1/NRSE sites found in the regulatory regions of numerous neuronal genes. Dysregulation of REST and its targeted genes has been reported in different cell and mouse HD models, as well as in biopsies from human patients. In this work, we characterized transcriptional dysregulation associated with REST in two different HD mouse models and assessed the therapeutic effect of interfering with REST function by overexpressing a dominant-negative form (DN:REST). We show that delivery of DN:REST in the motor cortex restores brain-derived neurotrophic factor (BDNF) mRNA and protein levels by reducing endogenous REST occupancy at the Bdnf locus. Similarly, expression of other REST-regulated genes such as Synapsin I (Syn1), Proenkephalin (Penk1) and Cholinergic receptor muscarinic 4 (Chrm4) were restored to normal levels while non-REST-regulated genes were unaffected. This is the first study conducted to investigate REST's role in vivo in a neurodegenerative disease. Our data show that DN:REST in motor cortex reversed RESTs repressive effects on target genes. However, the lack of therapeutic effect on motor function suggests that a more widespread rescue of REST-regulated sites in the affected brain regions may be necessary.

Mitochondrial role in ischemia-reperfusion of rat hearts exposed to high-K+ cardioplegia and clonazepam: energetic and contractile consequences.

The role of the mitochondrial Na/Ca-exchanger (mNCX) in hearts exposed to ischemia-reperfusion (I/R) and pretreated with cardioplegia (CPG) was studied from a mechano-calorimetric approach. No-flow ischemia (ISCH) and reperfusion (REP) were developed in isolated rat hearts pretreated with 10 micromol/L clonazepam (CLZP), an inhibitor of the mNCX, and (or) a high K+ - low Ca2+ solution (CPG). Left ventricular end diastolic pressure (LVEDP), pressure development during beats (P), and the steady heat release (Ht) were continuously measured and muscle contents of ATP and PCr were analyzed at the end of REP. During REP, Ht increased more than P, reducing muscle economy (P/Ht) and the ATP content. CPG induced an increase in P recovery during REP (to 90% +/- 10% of preISCH) with respect to nonpretreated hearts (control, C, to 64% +/- 10%, p < 0.05). In contrast, CLZP reduced P recovery of CPG-hearts (50% +/- 6.4%, p < 0.05) and increased LVEDP in C hearts. To evaluate effects on sarcoplasmic reticulum (SR) function, ischemic hearts were reperfused with 10 mmol/L caffeine -36 mmol/L Na (C - caff - low Na). It increased LVEDP, which afterwards slowly relaxed, whereas Ht increased (by about 6.5 mW/g). CLZP sped up the relaxation with higher DeltaHt, C - caff - low Na produced higher contracture and lower Ht in perfused than in ischemic hearts. Values of DeltaHt were compared with reported fluxes of Ca2+-transporters, suggesting that mitochondria may be in part responsible for the DeltaHt during C - caff - low Na REP. Results suggest that ISCH-REP reduced the SR store for the recovery of contractility, but induced Ca2+ movement from the mitochondria to the SR stores. Also, mitochondria and SR are able to remove cytosolic Ca2+ during overloads (as under caffeine), through the mNCX and the uniporter. CPG increases Ca2+ cycling from mitochondria to the SR, which contributes to the higher recovery of P. In contrast, CLZP produces a deleterious effect on ISCH-REP associated with higher heat release and reduced resynthesis of high energy phosphates, which suggests the induction of mitochondrial Ca cycling and uncoupling.

Dioxygenation of naphthalene by Pseudomonas fluorescens N3 dioxygenase: optimization of the process parameters.

The bioconversion of naphthalene to the 1,2-dihydro-1,2-dihydroxy derivative was performed in good yield using an Escherichia coli recombinant strain carrying Pseudomonas fluorescens N3 dioxygenase. However, the efficiency of such transformation is affected by many process parameters, and their optimization is essential to the scaling up of the process. The following process parameters were considered for optimization: cell concentration together with the corresponding glucose concentration (DCW/L); pH of medium; temperature; stirring speed; air flow; substrate concentration; Fe(2+) concentration; microelements concentration; reaction volume. We used a two-step multivariate experimental design to select important variables and assign them optimal values. The most significant parameters were selected by adopting a Plackett-Burman design, and were then correlated, using a full factorial design, with the experimental results. The experimental results illustrate that the optimized process of recombinant whole cell biotransformation in two-liquid phase systems enhances the naphthalene dihydrodiol yield threefold. This biotransformation opens the way to future experiments involving different substrates.

Comparison of two cysteine endopeptidases from latices of Morrenia brachystephana Griseb. and Morrenia odorata (Hook et Arn.) Lindley (Asclepiadaceae).

The properties of morrenain b II, a proteinase isolated from the latex of Morrenia brachystephana, were compared with those of morrenain o II, a proteinase obtained from the latex of Morrenia odorata. Both peptidases were purified to homogeneity by acetone precipitation followed by cation exchange chromatography. The enzymes have pI values higher than 9.3 and similar molecular masses (close to 26 kDa) as determined by SDS-PAGE. They display maximum proteolytic activity within an alkaline pH range, and also exhibit esterolytic activity. The N-terminal sequences of morrenain o II and morrenain b II show a high degree of homology between each other and to other cysteine plant proteinases.

What is the cost of a healthy diet in terms of achieving RDAs?

OBJECTIVE: Assessing how the Italian average food pattern would be affected in terms of consumption structure and expenditure by the adoption of nutritional prescriptions. DESIGN: A linear programming model with nutritional and food habits constraints was employed to generate a pattern following recommended daily allowances (RDAs) and nutritional guidelines provided for the Italian population. SETTING: Food consumption data from ISTAT Household Budget Survey of Italian population. SUBJECTS: Italian families investigated by the Family Budget Survey of the National Institute of Statistics. RESULTS: Compared to actual behaviour, the pattern generated by the model implies an increased consumption of vegetables, pasta, rice and fresh fish, and a decreased consumption of meats, bread, sugars and cakes, and especially fats and oils. At given prices, total expenditure is lower than actual expenditure. CONCLUSIONS: Differences between actual behaviour and the generated pattern are consistent with long-term trends in food consumption. The adoption of RDAs is unlikely to result in an increased food expenditure.

Comparison of dirithromycin and penicillin for treatment of streptococcal pharyngitis.

In the treatment of group A beta-hemolytic streptococcal pharyngitis, penicillin is the drug of choice and erythromycin is the alternative. In a double-blind, randomized study, dirithromycin, a new macrolide, was compared with penicillin for the treatment of streptococcal pharyngitis. Of 121 patients who were treated with dirithromycin, 96.7% manifested a favorable clinical response, and of 136 patients treated with penicillin, 94.2% manifested a favorable clinical response. Streptococci were eradicated from the pharynges of 85.3% of 116 dirithromycin-treated patients and 82.5% of 126 penicillin-treated patients who returned for follow-up. There were no statistically significant differences in efficacy between the two groups. The incidence of abdominal symptoms was higher in dirithromycin-treated patients. Being as efficacious as penicillin and having the advantages over erythromycin of once-daily dosing and the lack of drug interactions, dirithromycin is an alternative to penicillin in the treatment of streptococcal pharyngitis for patients 12 years of age and older.

Clinical efficacy of dirithromycin in patients with bacteremic pneumonia.

Dirithromycin is a new macrolide antimicrobial drug with a long half-life (44 hours) that reaches high tissue concentrations, thus permitting once-daily oral dosing and shorter courses of therapy. Soon after absorption, dirithromycin enters the tissue so rapidly that serum concentrations are comparatively low. It could be hypothesized that these low serum levels could endanger the outcome in patients with bacteremic pneumonia. We reviewed the database on dirithromycin pneumonia (consisting of 1108 patients randomized to receive dirithromycin or erythromycin in two double-masked trials) to ascertain its efficacy in patients with community-acquired pneumonia and concomitant bacteremia. Fourteen (2.5%) of 555 dirithromycin-treated patients and 10 (1.8%) of 553 erythromycin-treated patients had bacteremia. A favorable clinical response posttherapy was observed in 92.3% and 88.9% of these patients with a response assigned, respectively. Overall, favorable response rates were comparable between the two groups in the bacteremic subsets: patients with pneumococcal bacteremia, patients with nonbacteremic pneumococcal pneumonia, and all patients enrolled with acute pneumonia who had a posttherapy clinical response. In the treatment of patients with mild or moderate community-acquired pneumonia, including those with unsuspected and incidental bacteremia, dirithromycin is an effective macrolide antimicrobial drug.

Tonsillar tissue penetration of dirithromycin after multiple doses.

Dirithromycin is a new macrolide antibiotic that is effective against group A beta-hemolytic streptococcal pharyngotonsillitis. This prospective, multicenter, randomized study compared the serum and tonsil tissue concentrations of erythromycylamine (to which dirithromycin is rapidly converted by nonenzymatic hydrolysis during absorption) and erythromycin after 5- and 10-day regimens of dirithromycin and erythromycin, respectively. Thirty-nine patients undergoing elective tonsillectomy but without active tonsillitis were assigned in randomized fashion to receive dirithromycin 500 mg orally once daily (n = 22) or erythromycin base 250 mg orally four times daily (n = 17). Data from 12 patients receiving dirithromycin and 10 receiving erythromycin were eligible for analysis. Mean serum concentrations (+/-standard deviation) of erythromycylamine and erythromycin were 0.20 +/- 0.07 microgram/mL and 0.12 +/- 0.25 microgram/mL, respectively, after the 5-day regimen and 0.17 +/- 0.10 microgram/mL and 1.57 +/- 3.16 micrograms/mL, respectively, after the 10-day regimen. The mean serum concentration of erythromycin after 10 days was skewed by the data for one of the six patients in the group (concentration of > 8 micrograms/mL). Mean concentrations of erythromycylamine in tonsil tissue were 4.62 +/- 0.97 micrograms/ g after 5 days and 3.47 +/- 2.84 micrograms/g after 10 days. Concentrations in tonsillar tissue were undetectable in all patients given erythromycin for 5 days and in 4 of the 6 patients given erythromycin for 10 days. The high concentrations of erythromycylamine in tonsillar tissue agree with the clinical efficacy seen in the treatment of group A beta-hemolytic streptococcal tonsillopharyngitis with dirithromycin.

Isolation of Borrelia burgdorferi in Tuscany (Italy).

To evaluate the diffusion of Borrelia burgdorferi in Tuscani (Italy) 720 ticks were collected and subsequently cultured for Borrelia burgdorferi. A strain of Borrelia burgdoferi was isolated from one tick; this being the first such reported identification in Central-Southern Italy.

Cranial fractures associated with facial fractures: a review of mechanism, type, and severity of injury.

A 100-month retrospective review was undertaken to identify the population characteristics of patients with both facial and cranial fractures and to establish the relationships between them. A 4.4% incidence of cranial fractures was found in the 882 patients with facial fractures. These patients tended to be males (85%) between the ages of 16 and 30 years (54%) who were involved in motor vehicle accidents (64%). Patients with midfacial fractures predominated (70%), and these injuries were most frequently associated with fractures of the frontal (38%), sphenoid (24%), or temporal (22%) bones. No relationship was noted between mandibular fractures and cranial fractures. Midfacial fractures were related to individual cranial bone fractures by sutural attachment. The presence of cranial fractures did not play a role in the development of complications associated with facial fractures.

Use of multivariate analysis to compare antimicrobial agents on the basis of in vitro activity data.

Multivariate techniques such as principal component analysis or similar factor analysis help in analyses of the simultaneous interrelationships among several variables. A comparative multivariate analysis on the in vitro activities of eight antimicrobial agents, including the novel molecule daptomycin, is presented. Multivariate analysis detects components or factors and establishes connections among antimicrobial agents on the basis of their different levels of participation in each factor. The first principal component was dominated by vancomycin, teicoplanin, and rifampin (0.94344, 0.92792, and 0.72127, respectively). The second principal component showed strong effects from imipenem, gentamicin, and cephalothin (0.87922, 0.86126, and 0.68870, respectively). Daptomycin stood out alone in the third principal component (0.83983). The first three components defined 81.5% of the total variance and could easily be represented graphically in a three-dimensional scatter plot. In this graphic representation, the eight antimicrobial agents clustered in three different spatial regions; daptomycin occupied a separate spatial position. The use of multivariate analysis offers a different approach to determination of the in vitro activities of new antimicrobial agents and adds some new data on the relationships among different classes. Notwithstanding its limitations, the application of these methods in microbiology and drug development could be an additional tool for use in processing information.

Management of closed head injury in the patient with maxillofacial trauma.

Closed head injury (CHI) affects approximately one in five patients who sustain facial fractures. The effects can range from simple loss of consciousness to coma and death. The high incidence of CHI in the facial trauma population and the potential for mortality and neurologic morbidity make it a distinct concern of the practicing oral and maxillofacial surgeon. This article discusses the evaluation, monitoring, and management of CHI and includes special consideration of the treatment of maxillofacial injuries in these patients.

Safety profile of dirithromycin.

Dirithromycin is a recently developed oral antibiotic, and has been shown to be effective in the treatment of respiratory tract, skin and soft tissue infections. Dirithromycin is administered once daily which may contribute to patient compliance. In this paper we review the data from studies conducted in Europe, USA, Israel and South Africa over a six-year period to assess the safety and efficacy of dirithromycin in the treatment of a variety of acute infectious illnesses, and to compare it with structurally related antibiotics (erythromycin base, roxithromycin, and miocamycin) given orally. A total of 7437 patients have been enrolled from a total of 66 studies and trials, 4263 (57.3%) treated with dirithromycin and 3174 (42.7%) treated with a comparator antibiotic. Patients received either 500 mg dirithromycin (two tablets once daily), 1000 mg erythromycin base (250 mg qid), 300 mg roxithromycin (150 mg bid), or 1200 miocamycin (600 mg bid); the length of therapy ranged from 7 to 14 days. These studies have shown that dirithromycin has a safety profile similar to the comparator agents. The most frequently reported adverse events for both dirithromycin and comparator treatment groups were gastrointestinal in nature. The majority (99%) of adverse events reported from patients treated with dirithromycin were considered mild or moderate in severity. Early discontinuation of antibiotic therapy was infrequent (3-4%) in both treatment groups, and considered to be possibly drug-related in 2-3% of the population. The safety profile of dirithromycin in elderly patients was comparable to that recorded in the overall patient population. The incidence and nature of abnormal clinical laboratory evaluation were similar in dirithromycin and comparator groups. Notable alterations in laboratory tests of haematological or hepatic function were infrequent and were not associated with clinical manifestations. Routine monitoring of standard clinical laboratory tests in patients prescribed dirithromycin does not appear to be necessary.

Dirithromycin in the treatment of streptococcal pharyngitis.

A double-blind, double-dummy, randomized, multicentre study compared the safety and the efficacy of dirithromycin (two 250 mg tablets, once daily) to erythromycin base (four 250 mg tablets, four times daily) for ten days in the treatment of proven group A beta-haemolytic (GABHs) streptococcal pharyngitis. Five-hundred and fifty-three patients (265 dirithromycin, 288 erythromycin) were enrolled in the trial and analysed for efficacy and safety. Clinical and bacteriological evaluations were performed pre-therapy, during therapy (days 3-5), post-therapy (three to five days after completion of treatment), and late post-therapy (three to five weeks after treatment). All patients qualified for safety analysis. A negative pre-therapy culture was the predominant reason a patient did not qualify for analysis of efficacy. At post-therapy, favourable clinical responses (cure or improvement) were reported for 94.1% (159/169) of dirithromycin and 94.6% (158/167) of erythromycin patients who qualified for efficacy analysis. Post-therapy throat cultures were negative for GABH streptococci in 79.3% (134/169) of dirithromycin patients and 86.2% (144/167) of patients treated with erythromycin (P = 0.314). At late post-therapy 87.6% (134/153) of dirithromycin and 88.7% (134/151) of erythromycin patients reported a favourable clinical response; pathogens were absent in 69.9% (107/153) of dirithromycin and 86.1% (130/151) of erythromycin patients (P = 0.001). The adverse event profile of the two drugs was comparable although digestive and cutaneous adverse events were reported more frequently in the erythromycin treatment group. In this study, more dirithromycin patients had throat cultures positive for GABH streptococci at late post-therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical economics: a method for prospective health resource data collection.

Information about the economic benefit of new drugs is becoming increasingly important for formulary considerations, reimbursement policies and related considerations. Although economic benefits of drugs have been analysed and reported, the economic benefits of drugs have rarely been examined in the course of randomised therapeutic trials. We designed a modular survey instrument, the Resource Utilisation Survey (RUS), to collect economic outcomes in prospective trials. A pilot test of the RUS was conducted using clinical trial methods in a study of nizatidine versus placebo in preventing ulcers induced by nonsteroidal anti-inflammatory drugs. The purpose of the pilot study was to evaluate the RUS instrument and corresponding study design issues in clinical trials for either acute or chronic diseases. With the lessons learned from the pilot study, the RUS has been used successfully in other ongoing clinical trials.

A review of 100 closed head injuries associated with facial fractures.

One hundred closed head injuries associated with facial fractures treated over a 78-month period at a level I trauma center in Northeast Ohio were reviewed. The incidence of closed head injury in patients with facial fractures was 17.5%. Males suffered closed head injuries four times more often than females, and sustained severe intracranial injuries eight times as often. The 16- to 30-year age group predominated (59%). Although motor vehicle accidents were the most frequent cause of injury (61%), motorcycle accidents were associated with the most severe head injury. The mandible/midface fracture ratio (1.3:1) was almost half that of the non-head-injured population (2.1:1). Facial fracture complications were found to have a similar incidence (14%) as in the non-head-injured population, but were associated with more severe intracranial injuries.

Cervical spine fractures and maxillofacial trauma.

Cervical spine fractures are a serious complication of maxillofacial trauma because of the high potential for mortality and neurologic morbidity. This study reviewed 563 patients with facial fractures treated by the Division of Oral and Maxillofacial Surgery at a level I trauma center and identified 11 concomitant cervical spine fractures (2.0%). These patients were almost exclusively male (91%), white (100%), between 20 and 35 years of age (64%), involved in a motor vehicle accident (91%), with a concomitant mandibular fracture (91%). The examination, diagnosis, and management of patients with cervical spine injuries are discussed.