Variations in [3H]ouabain binding of porcine skeletal muscle associated with feeding.

The maximal ouabain binding capacity (Bmax) of longissimus dorsi muscle has been determined over a 24 h period in young pigs living at 12 degrees C. Animals were fed once a day and Bmax was estimated at 4, 8, 12 or 24 h after a large meal. Mean values of Bmax +/- S.E.M. (pmol ouabain/g wet wt muscle) of 548 +/- 78 and 556 +/- 57 at 4 and 24 h after feeding were significantly greater than those of 383 +/- 24 and 393 +/- 30 at 8 and 12 h after feeding (P less than 0.02). The extent to which these differences represent apparent or real changes in numbers of ouabain binding sites and, hence, in Na+,K(+)-ATPase concentration is discussed.     

Dissociation between plasma concentrations of thyroxine and insulin-like growth factor-I

The relation between plasma concentrations of thyroxine (T4) and insulin-like growth factor-I (IGF-I) has been examined in young, growing pigs under controlled conditions of energy intake. Compared with euthyroid controls, plasma levels of IGF-I were significantly elevated (P less than 0.005) both in hypothyroid animals on the same food intake and in hyperthyroid animals on double the food intake. There was however no increase in IGF-I in a hyperthyroid group on the control level of intake. Contrary to previous reports in which energy intake was not controlled, it is concluded that there is no simple correlation between plasma concentrations of T4 and IGF-I.     

Molecular characterization of Staphylococcus aureus isolated from children with adenoid hypertrophy: emergence of new spa types t7685 and t7692

Objective

Adenoids have been associated with the pathogenesis of acute, recurrent and chronic infectious diseases of the upper respiratory system and their hypertrophy is one of the most common causes of upper airway obstruction affecting children. In this study, the characteristics of Staphylococcus aureus isolates from patients who had undergone adenoidectomy were investigated via spa typing method.

Methods

A total of 113 children with adenoid hypertrophy who underwent adenoidectomy during September 2009 to November 2010, were included in the study. The isolates were identified to the species level as S. aureus using standard biochemical methods, following which the amplification and sequencing of the spa gene X region were carried out.

Results

S. aureus was found in the adenoid tissue of 26 (23%) patients. Out of the 26 S. aureus isolates, 5 (19%), 3 (11.5%) and 3 (11.5%) were resistant to tetracycline, erythromycin and oxacillin respectively. All the isolates were susceptible to vancomycin, rifampin, ciprofloxacin, gentamicin, mupirocin and quinupristin-dalfopristin and were typed using spa typing method. All the isolates were found to include 21 spa types, including two previously unreported types (t7685 and t7692). The most prevalent spa types were t7685 (11.5%), t230 (8%), t325 (8%) and t1149 (8%).

Conclusion

This study demonstrates that the prevalence rate of S. aureus in the adenoid tissue of the children assessed was 23%. An interesting point to note was the dominance of the spa type t7685 that has not been previously reported by other studies.     

Cloning,Expression, and Purification of Hyperthermophile α-Amylase from Pyrococcus woesei

Objectives

In an attempt α-amylase gene from Pyrococcus woesei was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY- α-amylase.

Methods

Escherichia coli BL21 used as a host and protein expression was applied using IPTG. SDS-PAGE assay demonstrated the 100 kDa protein. Amylolytic activity of proteins produced by transformed E. coli cells was detected by zymography, and the rate of active α-amylase with and without the intein tag in both soluble conditions and as inclusion bodies solubilized by 4M urea were measured.

Results

Amylolytic activity of ∼185,000 U/L of bacterial culture was observed from the soluble form of the protein using this system.

Conclusion

These results indicate that this expression system was appropriate for the production of thermostable α-amylase.     

Frequency of Alloicoccus otitidis, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae in children with otitis media with effusion (OME) in Iranian patients

Objective

To determine the presence of common bacterial agents of otitis media with effusion (OME), together with investigation these agent in the adenoid tissue and antimicrobial susceptibility pattern of isolated bacteria in Iranian children with OME.

Methods

Polymerase chain reaction (PCR) and bacterial culture methods were used for detection and isolation of Alloicoccus otitidis, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae in 63 middle ear fluid samples and 48 adenoid tissues from 48 OME patients. Fifteen patients were bilaterally affected. Antimicrobial susceptibility of all bacterial isolates were determined by disk agar diffusion (DAD) method.

Results

Bacteria were isolated from 47% (n = 30) of the middle ear fluid samples and 79% (n = 38) of the adenoid tissue specimens in OME patients. A. otitidis was the most common bacterial isolated from the middle ear fluid 23.8% by culture and 36.5% by PCR method. S. pneumoniae was the most prevalent pathogen (35.5% and 31.2% by culture and PCR) in the adenoid tissues. In 10 patients the same organisms were isolated from the middle ear fluid and adenoid tissue. Antimicrobial susceptibility pattern showed taht most isolates of bacteria were sensitive to ampicillin, Amoxicillin/Clavulanate and fluoroquinolones.

Conclusion

The present study, being the first report on the isolation of A. otitidis by culture method in Iran and Asian countries, shows that A. otitidis is the most frequently isolated bacterium in Iranian children having otitis media with effusion. In this study A. otitidis, S. pneumoniae, H. influenzae and M. catarrhalis are the major bacterial pathogens in patients with OME and we found that ampicillin and Amoxicillin/Clavulanate have the excellent activity against bacterial agents in Iranian children with OME.     

Evaluation of biofilm production and characterization of genes encoding type III secretion system among Pseudomonas aeruginosa isolated from burn patients

Pseudomonas aeruginosa is one of the common pathogenic causes of serious infections in burn patients throughout the world. Type III secretion toxins are thought to promote the dissemination of P. aeruginosa from the site of infection, the bacterial evasion of the host immune response and inhibition of DNA synthesis leading to host cell death. A total of 96 isolates of P. aeruginosa were collected from wound infections of burn patients, from April to July 2010. Antimicrobial susceptibility of the isolates were determined by disk agar diffusion method. Polymerase chain reaction (PCR)-based method was used for targeting the genes encoding the type III secretion toxins.