Structural and functional characterization of recombinant human growth hormone isolated from transgenic pig milk.

This study aimed to establish and reproduce transgenic pigs expressing human growth hormone (hGH) in their milk. We also aimed to purify hGH from the milk, to characterize the purified protein, and to assess the potential of our model for mass production of therapeutic proteins using transgenic techniques. Using ~15.5 L transgenic pig milk, we obtained proteins with ≥ 99% purity after three pre-treatments and five column chromatography steps. To confirm the biosimilarity of our milk-derived purified recombinant hGH (CGH942) with commercially available somatropin (Genotropin), we performed spectroscopy, structural, and biological analyses. We observed no difference between the purified protein and Genotropin samples. Furthermore, rat models were used to assess growth promotion potential. Our results indicate that CGH942 promotes growth, by increasing bone development and body weight. Toxicity assessments revealed no abnormal findings after 4 weeks of continuous administration and 2 weeks of recovery. The no-observed-adverse-effect level for both males and females was determined to be 0.6 mg/kg/day. Thus, no toxicological differences were observed between commercially available somatropin and CGH942 obtained from transgenic pig milk. In conclusion, we describe a transgenic technique using pigs, providing a new platform to produce human therapeutic proteins.

Anesthetic requirements and stress hormone responses in spinal cord-injured patients undergoing surgery below the level of injury.

Neuraxial anesthesia decreases the minimum alveolar concentration. We determined the effects of spinal cord injury (SCI) on sevoflurane requirements and stress hormone response. Twenty-two chronic SCI patients undergoing surgery below the level of the injury were enrolled in the study, and 15 patients without cord injury served as control patients. Bispectral index score was maintained at 40-50. Measurements included end-tidal sevoflurane concentrations, systolic arterial blood pressure, heart rate, and plasma catecholamine and cortisol concentrations. During surgery, systolic arterial blood pressure, heart rate, and Bispectral index were comparable between SCI and control groups. However, end-tidal sevoflurane concentration was significantly smaller in the SCI (0.81%-1.06%) versus control (1.28%-1.31%) patients. In the control group, plasma norepinephrine and cortisol concentrations were significantly increased during and 1 h after surgery compared with awake baseline values. In the SCI group, the sympathoadrenal and cortisol responses were virtually abolished. We conclude that SCI reduces the anesthetic requirement by 20%-39% during surgery below the level of injury, in association with blunted sympathoadrenal and cortisol responses.

Marine bacteria as a leading cause of necrotizing fasciitis in coastal areas of South Korea.

Necrotizing fasciitis is a life-threatening soft-tissue infection. It is useful to know the etiology of this disease in each community and region because this information can facilitate early diagnosis and treatment. We reviewed medical records of 217 consecutive patients with necrotizing fasciitis in South Korea. Etiologic organisms were documented in 171 (78.8%) patients. Gram-negative marine bacteria were the most common organisms (68.4%). Seasonal variation in incidence occurred during warmer periods. Liver cirrhosis and alcoholism were common underlying illnesses. Gastrointestinal symptoms were more common in persons infected with gram-negative marine bacteria (56.4% versus 29.6%). Gram-negative bacteria were the predominant pathogens in multiple site involvement. Acute Physiology and Chronic Health Evaluation II scores, marine bacteria, and medical treatment without surgery affected mortality. When necrotic areas included less than three extremities, fasciotomy improved the survival rate. In coastal areas, gram-negative marine bacteria are the most common pathogens associated with necrotizing fasciitis. Fasciotomy can improve survival of patients with necrotic areas that involve less than three sites in necrotizing fasciitis.

Ketanserin, a 5-HT2 antagonist, directly inhibits the ATP-sensitive potassium channel in mouse ventricular myocytes.

The effects of ketanserin, a 5-HT2 antagonist, on the ATP-sensitive K+ (K(ATP)) channels were studied in mouse ventricular myocytes using patch clamp technique. Under the whole-cell voltage clamp conditions, ketanserin (1-100 microM) reversibly inhibited pinacidil-induced K(ATP) current in a concentration-dependent fashion with a Ki value of 9.36 microM and the Hill coefficient was 0.67. This inhibition was developed even with the presence of 5-hydroxytryptamine (100 microM) in the bath. Prazosin, a selective alpha1-antagonist, also failed to mimic the effect of ketanserin. Ketanserin did not affect the channel activity in inside-out configuration under the ATP-free internal solution. Furthermore, ketanserin applied to the external solution did not affect the pinacidil-induced channel activity in the cell-attached patches, but did inhibit it when applied into the pipette. These results suggest that the inhibitory action of ketanserin observed in this study was probably due to a direct action on the K(ATP) channel rather than to an action through the 5-HT2 receptor or alpha1-adrenoceptor blockade, and that the antiarrhythmic activity of ketanserin against cardiac arrhythmias induced in the ischemic/reperfused heart is at least in part attributable to its inhibition of the K(ATP) channel.