The discovery of (1R, 3R)-1-(3-chloro-5-fluorophenyl)-3-(hydroxymethyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile, a potent and selective agonist of human transient receptor potential cation channel subfamily m member 5 (TRPM5) and evaluation of as a potential gastrointestinal prokinetic agent.

This publication details the discovery of a series of selective transient receptor potential cation channel subfamily M member 5 (TRPM5) agonists culminating with the identification of the lead compound (1R, 3R)-1-(3-chloro-5-fluorophenyl)-3-(hydroxymethyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile (39). We describe herein our biological rationale for agonism of the target, the examination of the then current literature tool molecules, and finally the process of our discovery starting with a high throughput screening hit through lead development. We also detail the selectivity of the lead compound 39 versus related family members TRPA1, TRPV1, TRPV4, TRPM4 and TRPM8, the drug metabolism and pharmacokinetics (DMPK) profile and in vivo efficacy in a mouse model of gastrointestinal motility.

Is there a learning curve in static computer-assisted implant surgery? A prospective clinical study.

Static computer-assisted surgery (s-CAS) has been introduced to improve the results of implantology. A prospective cohort study was conducted following the STROBE guidelines to determine the presence of a learning curve in s-CAS. Six partially and six totally edentulous patients were treated by two surgeons experienced in implantology but completely inexperienced in s-CAS. Preoperative and postoperative computed tomography scans were matched to assess coronal, apical, and angular deviation and the positioning error. The accuracy data were used to evaluate the learning curve. Fifty-six implants were inserted. In partially and totally edentulous patients, the mean (range; standard deviation) coronal deviation was 0.87 (0.34-1.27; 0.35) and 1.24 (0.72-2.67; 0.79); the mean apical deviation was 1.13 (0.48-1.63; 0.39) and 1.52 (0.88-3.84; 1.15); the mean angular deviation was 2.63 (1.89-4.50; 0.98) and 3.59 (1.69-6.30; 1.65); and the mean positioning error was 0.80 (0.32-1.25; 0.35) and 1.14 (0.35-2.56; 0.77), respectively. A typical 'learning curve' effect was not identified for s-CAS.

Implant-Supported Rehabilitation Using GBR Combined with Bone Graft on a Reconstructed Maxilla with the Fibula Free Flap.

Alveolar ridge augmentation procedures allow restoring jaw defects due to teeth extractions, periodontal diseases, trauma, or outcomes from a previous surgery. This case report describes a patient suffering from Fibrous Dysplasia of the right upper maxilla surgically reconstructed by fibula free flap. In 2003, four dental implants were placed in the 1.2, 1.3, 1.5, and 1.6 areas. Twelve years later, the onset of peri-implantitis led to the failure of osseointegration with consequent thinning of the fibula flap. To avoid the risk of fracture and to restore the bone volumes necessary for a new implant-prosthetic rehabilitation, we used heterologous biomaterials in combination with a non-reabsorbable membrane, according to the Guided Bone Regeneration (GBR) technique. GBR was performed using the Equimatrix® natural bone mineral matrix, Cytoplast™ Ti-150, a non-reabsorbable titanium-reinforced membrane, and four fastening screws to pin the membrane. After six months, the membrane was removed and two Zimmer® implants 3.7 × 13 mm were placed in the 1.1 and 1.2 areas. A fixed implant-supported prosthesis with a custom-milled titanium bar screwed to the implants was made. Computed tomography (CT) six months after GBR showed a good bone regeneration of 1.5 cm mesiodistal (MD), 1.8 cm buccopalatal (BP), and 2.8 cm in height. The main difficulty of this clinical case concerns the low predictability of success of GBR on a maxillary reconstructed area with a free fibula flap: there is no previous evidence in the literature. Clinical and radiographic exams nowadays show that there is no macroscopic bone reabsorption; however, further research is needed to obtain more information.

Timing for implant placement in patients treated with radiotherapy of head and neck.

BACKGROUND: Aim of this retrospective study was to evaluate the most appropriate timing for the implant placement in oral cancer patients treated with radiation therapy. MATERIALS AND METHODS: We collected data for 17 subjects (10 females, 7 males; mean age: 51 ± 19) with malignant oral tumors who had been treated with ablative surgery and radiation therapy of head and neck. The radiotherapy was set in accordance with the NCCN guidelines. Every patient received dental implant rehabilitation between 2014 and 2016. A total of 84 titanium implants were placed, at a minimum interval of 12 months, in irradiated residual bone. Every patient underwent a minimum follow-up of 12 months. RESULT: Implant loss was dependent on the position and location of the implants (P = 0.05-0.1). Moreover, implant survival was dependent on the total dose of radiation and, mostly, on the timing of implant insertion after the end of the radiation therapy. This result was highly statistically significant (P < 0.01). Whether the implant was loaded is another highly significant (P < 0.01) factor determining survival. We observed significantly better outcomes when the implant was placed at least after 14 months and not loaded until at least 6 months after placement. CONCLUSION: Although the retrospective design of this study could be affected by selection and information biases, we conclude that a delayed implant placement after the end of the radiation therapy and a delayed loading protocol will give the best chance of implant osseointegration, stability and, ultimately, effective dental rehabilitation.

Identification of TEM-135 ß-lactamase in Neisseria gonorrhoeae strains carrying African and Toronto plasmids in Argentina.

One hundred forty-three penicillinase-producing Neisseria gonorrhoeae (PPNG) isolates obtained in Argentina from 2008 and 2012 were examined to detect blaTEM-135 genes and to investigate plasmid profiles and multiantigen sequence types. Forty-two PPNG isolates were found to carry TEM-135, and two contained a new TEM derivative characterized as TEM-220. The blaTEM-135 allele was carried by the Toronto/Rio and African plasmids. Molecular epidemiology revealed that two blaTEM-135 isolates were related to previously described isolates from Thailand and China, indicating a common evolutionary origin.

Recombinant AgB8/1 ELISA test vs. commercially available IgG ELISA test in the diagnosis of cystic echinococcosis.

The diagnosis and clinical management of cystic echinococcosis (CE) rely on imaging and serology, the latter still having a complementary role as its accuracy in assessing cyst viability is unsatisfactory. We used an experimental IgG ELISA test based on the recombinant antigen rEgAgB8/1 cloned from Echinococcus granulosus to differentiate active from inactive/cured CE infection, comparing its performance to that of a commercially available ELISA test used routinely in our hospital laboratory. Both tests were performed on sera from 88 patients with hepatic echinococcal cysts, grouped according to cyst stage based on ultrasonographical morphology, and on 17 patients surgically treated for echinococcosis and 18 patients with nonparasitic hepatic cysts included as controls. Tests' performances did not differ significantly, but the overall concordance between tests drastically dropped when groups were analysed separately. Further longitudinal studies should evaluate whether these discrepancies reflect the different ability of either test to predict the evolution of cysts over time. Although the recombinant-AgB8/1-based ELISA test seems to have no clinical advantage over the commercially available ELISA test in the assessment of hepatic CE cyst viability, the easiness of production and reproducibility of high-quality recombinant antigens makes rEgAgB8/1 a valid candidate for use in CE ELISA diagnostic tests.

Molecular characterization of Echinococcus granulosus in south-eastern Romania: evidence of G1-G3 and G6-G10 complexes in humans.

Clin Microbiol Infect ABSTRACT: Echinococcus granulosus is the aetiological agent of cystic echinococcosis (CE), which is a public health problem in many eastern European countries, particularly in Romania, where the infection causes a high number of human and animal cases. To shed light on the transmission patterns of the parasite, we performed a genotyping analysis on 60 cyst samples obtained from patients who live in south-eastern Romania and who underwent surgery for liver or lung CE. DNA was extracted from the endocysts or the cyst fluids, and fragments of cytochrome c oxidase subunit 1 and NADH dehydrogenase subunit 1 mitochondrial genes (cox1 and nd1, respectively) were amplified by PCR and sequenced. We found that most of the samples analysed (59/60) belonged to the G1-G3 complex (E. granulosus sensu stricto), which contains the most widespread and infective strains of the parasite. We also identified the first human patient infected by a non-G1-G3 genotype of E. granulosus in this country. As the DNA sequence of this cyst sample showed maximum homology with the G6-G10 complex (Echinococcus canadensis), this is, in all likelihood, a G7 genotype, which is often found in pigs and dogs in most countries of eastern and south-eastern Europe.

The influence of smoking and surgical technique on the accuracy of mucosa-supported stereolithographic surgical guide in complete edentulous upper jaws.

BACKGROUND: The accuracy of a stereolithographic surgical guide depends on several variables that can affect at any time from the software-planning stage to the surgical field. AIM: The purpose of this study was to evaluate the accuracy of implants inserted using a single mucosa-supported stereolithographic surgical guide determining also the influence of surgical technique (fixed and not-fixed) and smoking on the deviation parameters. MATERIALS AND METHODS: 11 patients, totally edentate in the upper arch and needed an implant-prosthetic rehabilitation, were selected. 95 implants were planned and inserted. The pre- and post-operative CT images were compared using software. RESULTS: Global, coronal (Mean: 1.65; Range: 3.00-0.13; SD:0.56) and apical (Mean: 2.15; Range: 4.23-0.34; SD:0.81), and angular (Mean: 4.62; Range: 15.25-0.28; SD: 2.74) deviation values were determined. The mean values of mucosa thickness in smokers and nonsmokers patients were 4.53 mm and 3.42 mm respectively (p < .05). The accuracy data showed a better result for the angular deviation when the surgical template was fixed (p = .002) and a better global apical deviation in the nonsmokers (p <. 05). CONCLUSIONS: It is essential, especially in smoker patients, to respect a minimum safety distance of 3 mm from limiting anatomic structures.

rKv1.2 overexpression in the central medial thalamic area decreases caffeine-induced arousal.

The voltage-gated potassium channel Kv1.2 belongs to the shaker-related family and has recently been implicated in the control of sleep profile on the basis of clinical and experimental evidence in rodents. To further investigate whether increasing Kv1.2 activity would promote sleep occurrence in rats, we developed an adeno-associated viral vector that induces overexpression of rat Kv1.2 protein. The viral vector was first evaluated in vitro for its ability to overexpress rat Kv1.2 protein and to produce functional currents in infected U2OS cells. Next, the adeno-associated Kv1.2 vector was injected stereotaxically into the central medial thalamic area of rats and overexpression of Kv1.2 was showed by in situ hybridization, ex vivo electrophysiology and immunohistochemistry. Finally, the functional effect of Kv1.2 overexpression on sleep facilitation was investigated using telemetry system under normal conditions and following administration of the arousing agent caffeine, during the light phase. While no differences in sleep profile were observed between the control and the treated animals under normal conditions, a decrease in the pro-arousal effect of caffeine was seen only in the animals injected with the adeno-associated virus-Kv1.2 vector. Overall, our data further support a role of the Kv1.2 channel in the control of sleep profile, particularly under conditions of sleep disturbance.

Ex vivo assessment of serum cytokines in patients with cystic echinococcosis of the liver.

To investigate the usefulness of serum cytokine levels in the diagnosis of active cystic echinococcosis, we evaluated the cytokine profile of patients with hepatic cystic echinococcosis in different cyst stages, CE 1-2 (active), CE3a-3b (transitional) and CE4-5 (inactive). Ex vivo assessment of Th1 (IL12, TNFalpha) and Th2 (IL4, IL10) cytokines in sera was carried out using ELISA. Percentages of positive samples and median levels of IL12, TNFalpha and IL10 did not differ significantly between groups. However, patients with CE3b cysts, a stage clinically unresponsive to treatments, had statistically significantly higher median levels of IL4 and percentage of positive samples for IL4. We conclude that the analysis of serum cytokine dosage, at least in its present form, is not useful as a marker of cyst activity. However, our results support recent findings suggesting the chronic activity of CE3b cysts and suggest that this might be partly because of a skewed Th2 response.

In vitro and in vivo synergy of levofloxacin or amikacin both in combination with ceftazidime against clinical isolates of Pseudomonas aeruginosa.

The aim of the present study was to explore the antibacterial activity of the levofloxacin (LVX) and ceftazidime (CAZ) combination compared with the amikacin (AMK)/CAZ combination against Pseudomonas aeruginosa. Minimum inhibitory concentrations (MICs) were determined according to NCCLS. FIC indices (Fl) were calculated by the checkerboard technique. CAZ combined with LVX or AMK yielded Fls indicating synergism (Fl < or = 0.5) for 71/102 (69.6%) and 81/102 (79.4%) (p = 0.108), indifference (FI > 0.5-4) for 24/102 (23.5%) and 12/102 (11.7%) (p = 0.027), and antagonism (Fl > 4) for 7/102 (6.8%) and 9/102 (8.8%) (p = 0.602) strains, respectively. In vivo, CAZ/LVX was as bactericidal as CAZ/AMK combination. Our results support the potential role of LVX as an alternative to AMK in the combination therapy with CAZ in the treatment of P. aeruginosa severe infections. Anyway, further investigations and clinical trials are awaited until any definitive conclusions can be drawn.

Development of a lower body negative pressure box with an environmental control system for physiological studies.

A headward fluid shift occurs during microgravity exposure, which causes the cardiovascular adaptive syndrome. Different countermeasures have been proposed to decrease its symptomatology, like the application of lower body negative pressure (LBNP). A LBNP box with an environment control system was developed, aiming to improve features of LBNP boxes used worldwide. It consists of five carbon steel ribs in the shape of a cylinder, which is wrapped with high pressure resistant and transparent vinyl. Inner and outer-wheeled trolleys can comfortably and easily move the subject in and out of the box. A custom-made skirt is secured around the subject's waist by an adjustable belt. The other end is secured between two window-type wooden structures, which seal the LBNP box. Inlet and an outlet valves connect the external to the internal environment of the chamber and tube system allows air to circulate gently. Electronic sensors are used to adjust the airflow keeping a pre-set negative pressure without changing humidity and temperature inside the box. Structural, pressure profile and leaking tests were performed with successful results. The improvements of the present LBNP box have substantially decreased the undesirable side effects of uncontrolled environment conditions during rapid pressure changes, and increased test subjects' comfort.

[Non fermentative gram negative bacilli isolated in a hospital laboratory]. / Bacilos gramnegativos no fermentadores (BGNNF) aislados en un laboratorio hospitalario.

OBJECTIVE: To report a prospective study on non fermentative gramnegative bacilli, excluded Pseudomonas aeruginosa, isolated at Dr. Julio C. Perrando Hospital in Resistencia (Argentina). The goal of this study was to know their frequency and antimicrobial susceptibility. MATERIAL AND METHODS: For bacterial identifications we used biochemical tests. RESULTS: The greatest percentages of non fermentative gramnegative bacilli isolates were found in blood samples (25%), respiratory secretions and urine (23.9%). Acinetobacter baumannii (34.7%), Pseudomonas fluorescens/Pseudomonas putida (15.2%), Stenotrophomonas maltophilia (9.7%) and Burkholderia cepacia (8.7%) were the non fermentative gramnegative bacilli species most commonly isolated. Distribution of microorganism strains according to samples and area is also assessed. Antimicrobial sensitivity of most commonly isolated non fermentative gramnegative bacilli strain analyzed. CONCLUSIONS: We concluded that non fermentative gramnegative bacilli are most frequently present in hospitalized that in outpatients and antibacterial therapy must be provided according to bacteriological information.

Competence for genetic transformation in encapsulated strains of Streptococcus pneumoniae: two allelic variants of the peptide pheromone.

The nucleotide sequence of comC, the gene encoding the 17-residue competence-stimulating peptide (CSP) of Streptococcus pneumoniae (L. S. Havarstein, G. Coomaraswamy, and D. A. Morrison, Proc. Natl. Acad. Sci. USA 92:11140-11144, 1995) was determined with 42 encapsulated strains of different serotypes. A new allele, comC2, was found in 13 strains, including the type 3 Avery strain, A66, while all others carried a gene (now termed comC1) identical to that originally described for strain Rx1. The predicted mature product of comC2 is also a heptadecapeptide but differs from that of comC1 at eight residues. Both CSP-1 and CSP-2 synthetic peptides were used to induce competence in the 42 strains; 48% of the strains became competent after the addition of the synthetic peptide, whereas none were transformable without the added peptides.