Müllerian inhibiting substance (MIS), a Sertoli cell-derived glycoprotein and member of the transforming growth factor-beta supergene family, plays a key down-stream role in mammalian sex determination. Identification of a receptor for MIS has now been achieved in a MIS-responsive human vulvar carcinoma cell line, A431, using fluorescein isothiocyanate labeling of recombinant human MIS (FITC-rhMIS) and RRAs with iodinated carboxy-terminal rhMIS. Confocal fluorescence microscopy of A431 cells incubated on ice with 30-nM concentrations of covalently bound FITC-rhMIS reveals specific punctate cell surface fluorescent signal. Cytosolic fluorescent signal is seen after incubation at 37 C for 1 h as well as occasional apparent perinuclear accumulation. FITC-rhMIS coincubated with molar excesses of unlabeled rhMIS in A431 cells eliminates cell surface and cytosolic fluorescent uptake. Double label experiments with FITC-rhMIS and tetramethyl rhodamine isothiocyanate epidermal growth factor establish separate binding of each ligand, displaceable, respectively, by large molar excesses of unlabeled rhMIS or epidermal growth factor. RRAs reveal a single, high affinity (Kd, 5.8 nM), saturable, low abundance binding species for carboxy-terminal rhMIS. Solubilized supernatants of A431 whole cells cross-linked with 125I-carboxy-terminal rhMIS identify a band with a mol wt of 88,000 on electrophoresis and autoradiography. This identification of a MIS receptor in A431 cells now permits the design of affinity purification protocols using rhMIS, followed by direct protein microsequencing.
Purpose: This study performed a comparative evaluation of nutritional condition's improvement and clinical effects in accordance with the Nutrition Support Team (NST) consultation compliance of critically ill pediatric patients. Methods: The medical records of 64 critically ill pediatric patients (2 to 18 years old), who were officially referred to a NST consultant in pediatric intensive care unit from January to August 2015, were reviewed. The patients were divided into 2 groups according to the compliance of NST consultation answers. The total delivered/required caloric and protein ratio, weight, serum total protein, serum albumin, hemoglobin, and hematocrit were compared. Results: According to the NST consultation answer, 'nutrition support increase' occupied the largest proportion at 38.5%; 'maintenance' and 'decrease' accounted for 35.7% and 18.2% respectively. The NST compliance group and non-compliance group were 20 and 14 patients, respectively. Although total delivered/required caloric ratio was significantly increased in the NST compliance group (19.7%, P=0.036), there was no significant difference in the NST non-compliance group (5.1%, P=0.692). The total delivered/required protein ratio was increased (15.1%, P=0.163) in the NST compliance group and decreased (−4.7%, P=0.774) in the NST non-compliance group. The NST non-compliance group (−8.6%, P=0.219) was further reduced weight than the NST compliance group (−1.0%, P=0.820). The serum albumin was significantly increased in the NST compliance group (13.1%, P=0.003), but there was no difference in the NST non-compliance group (7.1%, P=0.433). Conclusion: Although 56.7% of NST consultations were needed for nutritional interventions, a lower NST compliance (53.8%) is the limit of nutritional support. The NST compliance group was supplied adequately with more calories and protein than before consultation and a more improved nutritional status. Therefore, aggressive NST consultation can help increase the therapeutic effect by improving the nutritional status. This study will form the basis to seek ways to further enhance NST compliance.
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