Characterization of a trifluoroethanol-induced partially folded state of alpha-lactalbumin

The protein alpha-lactalbumin exists in a partially folded molten globule state at pH 2.0, the A state. This state is believed to be compact, possessing a similar amount of secondary structure to the native state but having a flexible tertiary structure comprised mainly of non-specific hydrophobic clustering of residues. Addition of trifluoroethanol (TFE) to bovine, human and guinea pig alpha-lactalbumin at pH 2.0 has been found in each case to induce a conformational transition in the A state as monitored by circular dichroism, nuclear magnetic resonance chemical shifts, and 1-anilinonaphthalene-8-sulphonate binding. The mid-point of this transition is near 15% (v/v) TFE and is effectively complete by 50% (v/v) TFE at 315 K. Far ultraviolet circular dichroism ellipticities at 208 nm and 220 nm, usually taken as a measure of the degree of helical character, are substantially more negative in the TFE state than in the A state. Furthermore, backbone amide protons protected from solvent exchange in the A state are generally at least as strongly protected in the TFE state; patterns of protection appear similar in the two states and include at least part of both the B and C alpha-helices. One major difference from the A state is nevertheless evident: the ability to bind the fluorescent probe 1-anilinonaphthalene-8-sulphonate, characteristic of molten globule states, is lost in the TFE state. Like the A state, the TFE state of alpha-lactalbumin shows little chemical shift dispersion of side-chain resonances. Extensive line broadening in the nuclear magnetic resonance spectra, characteristic of slow conformational averaging in the A state, is, however, much reduced in the TFE state. The line narrowing observed in the TFE state has made it possible to obtain directly sequence-specific assignments for about 25% of the 123 residues of bovine alpha-lactalbumin in 50% (v/v) TFE. Two helices are amongst regions of structure so far identified from short-range backbone nuclear Overhauser enhancement (NOE) connectivities in two-dimensional spectra of the TFE state. One of the helices (residues 86 to 96) corresponds to the C-helix in the native structure. The other (residues 35 to 41) corresponds, however, to a region of the sequence that is not helical in the native state. The partially folded state of alpha-lactalbumin formed in TFE, therefore, supports both native and non-native secondary structure in the absence of persistent long-range tertiary structure.     

Tissue-engineered nipple reconstruction

We describe a simple, effective approach to the creation of autologous tissue-engineered cartilage in the shape of a human nipple by injecting a reverse thermosensitive polymer seeded with autologous chondrocytes in an immunocompetent porcine animal model. A biodegradable, biocompatible copolymer of polyethylene oxide and polypropylene oxide (Pluronic F-127), which exists as a liquid below 4 degrees C and polymerizes to a thick gel when it is exposed to physiologic temperatures (body temperatures), was used as a vehicle for chondrocyte delivery and as a scaffold to guide growth. Autologous chondrocytes isolated from porcine auricular elastic cartilage and suspended in 30% (weight/volume) Pluronic F-127 were injected on the ventral surface of the pigs from which the cells had been isolated. A circumferential subdermal suture was used to support the contour of the implant and assist in its projection in the form of a human nipple. After 3 weeks, the skin over and surrounding the implant was tattooed to create the appearance of a human nipple-areolar complex. As controls, an equal number of injections were made using either cells alone (not suspended in hydrogel), or hydrogel alone. After 10 weeks, all specimens were excised and examined both grossly and histologically. Before harvesting, visual inspection of the tattooed chondrocyte-Pluronic F-127 hydrogel implant sites revealed that they closely resembled a human female nipple-areolar complex. Nodules were similar in size, shape, and texture to a human nipple at each injection site. Glistening opalescent tissue was surgically isolated from each implant site. Hematoxylin and eosin, safranine o, trichrome blue, and Verhoeff's stains of the experimental implants showed nodules with the characteristic histologic signs of elastic cartilage. Control injections of copolymer hydrogel alone exhibited no evidence of cartilage formation. Control injections of chondrocytes alone showed evidence of dissociated microscopic nodules of elastic cartilage.     

The effect of education on mortality among older Taiwanese and its pathways

The present research examines the impact of education on the mortality of older Taiwanese during a 4-year interval from April 1989 to April 1993. Data used for this study come from the Taiwan Survey of Health & Living Status of the Elderly (1989). The research decomposes the effect of education into the direct effect and the indirect effects by means of health status, health behaviors, and social relationships. We have shown that, of the total effect of educational attainment on the mortality of older Taiwanese, about 83% represents indirect influences by means of the 3 mediating factors, particularly health status. On the other hand, the magnitude of the direct effect, which might reflect influences of additional intervening variables on old-age mortality, is low and not statistically significant. The results demonstrate that the apparent strong effect of education on mortality among older Taiwanese can be accounted for parsimoniously through 3 major pathways.     

Effect of electroacupuncture on gastric acid secretion and gut hormones

21 patients with mild type of chronic superficial gastritis were selected in this study. The effect of electroacupuncture in Zhongwan (RM12), Neiguan (P6) and Sanyinjiao (Sp6) on gastric acid secretion, serum gastrin, plasma somatostatin, plasma motilin concentration and erythrocyte acetylcholinesterase (AchE) activity were observed. The results were as follows: There were significant decreases in gastric acid output, serum gastrin concentration and AchE activity (P < 0.05), but no significant changes in plasma somatostatin and motilin concentration (P > 0.05) after simultaneous electroacupuncture in Zhongwan, Neiguan and Sanyinjiao.     

Effects of high glucose culture on EGF effects and EGF receptors in the LLC-PK1 cells

OBJECTIVE: To study interns' perceptions of their learning during their rotation through a short stay unit (SSU). DESIGN: Case-based, qualitative research study. SETTING: A tertiary care pediatric hospital (The Children's Hospital, Boston, Mass). PARTICIPANTS: Ten interns who had worked in the SSU in the 3 months prior to June 1, 1995, and on a general medical team in the previous 12 months. INTERVENTION: None. MAIN OUTCOME MEASURES: In July 1995, the interns participated in focused, open-ended interviews lasting about 40 to 60 minutes to document their perceptions of their learning during their SSU rotation. The interviews were recorded on audiotape and transcribed prior to analysis. Data were analyzed to discern and categorize themes from the interns' responses. RESULTS: All interns responded favorably to their educational and learning experiences during their rotation through the SSU. Two major themes emerged: (1) the interns' learning, which was affected by the role of the attending physician, the organization and structure of the SSU, and the teaching strategies in the SSU; and (2) the interns' collaborative work with the nursing staff in the SSU, which affected patient care but did not facilitate the interns' learning. CONCLUSION: Clustering in the SSU of patients whose symptoms suggested straightforward diagnoses enhanced interns' educational experiences.     

Myxoid neurofibroma of the male breast: fine needle aspiration cytodiagnosis

The use of colloid agents to achieve hypervolemia in the prevention and treatment of postsubarachnoid hemorrhage (post-SAH) vasospasm is included in the standard of care at many institutions. Risk profiles are necessary to ensure appropriate use of these agents. In a series of 85 patients with recent aneurysmal SAH, 26 developed clinical symptoms of vasospasm. Fourteen of the 26 were treated with hetastarch for volume expansion while the other 12 received plasma protein fraction (PPF). Clinically significant bleeding pathologies were noted in six patients who received hetastarch as a continuous intravenous infusion. Hetastarch increased partial thromboplastin time from a mean of 23.9 seconds to a mean of 33.1 seconds (p < 0.001) in all patients who received infusions of this agent, while no effect was noted in the 12 patients who received PPF infusions. No other coagulation parameters were altered. This study shows an increase in coagulopathy with the use of hetastarch as compared with the use of PPF for the treatment of postaneurysmal vasospasm.     

Effect of captopril on antithrombus function of endothelium

The study involved 355 specimens of STD clinic patients collected from Beijing, Shantou and Wuhan, for the detection of Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum by polymerase chain reaction. The results showed that the positive rate of Neisseria gonorrhoeae being the highest in the patients attending STD clinics from the three cities. The detection rate of Neisseria gonorrhoeae in male patients was higher than in females. The positive rates of Chlamydia trachomatis and Ureaplasma urealyticum in the three cities differed from each other greatly. Polymerase chain reaction was suitable for clinical Jetection and epidemiological study of the three kinds of STD pathogens.     

Esophagogastrectomy: reoperation for complications

We investigated various factors related to secondary hyperparathyroidism among hemodialysis patients. Subjects were 26 patients (20 men and 6 women) ranging in age from 24 to 75, treated at an ordinary hemodialysis center throughout the year 1991. The serum inorganic phosphate level and serum intact PTH level varied greatly from 2.5 to 12.5 mg/dl and from 10 to 1,102 pg/ml (normal range, 10-60 pg/ml), respectively. All patients were classified according to the serum intact PTH level into two groups: those with less than 120 pg/ml (Group I) and those with 120 pg/ml or more (Group II). The patients in Group I responded well to hemodialysis and drug therapy, but those in Group II were likely to have secondary hyperparathyroidism. In all patients in Group II, the PTH level increased markedly in the summer. This may be attributed to insufficient ingestion of calcium, vitamins, and other nutrients due to loss of appetite.     

Influence of the blight of Morinda officinalis How on microscopic structure and constituents of the host

The results showed that microscopic structure in biennial roots, the contents of B and Mn in nutritive organ and beta-sitosterol content in roots, stems of Morinda officinalis were affected markedly. The total sugar content in the roots and aerial stems was decreased by 5.42%-15.29%.     

Effects of different natural stimulants on the activity of the rabbits' carotid chemoreceptor

The effects of different natural stimulants on the afferent unit discharge of rabbit carotid chemoreceptors were studied in the in vitro carotid body- sinus nerve preparation. A total of 32 chemoreceptive units were recorded. The results were as follows: (1) Of the 32 units, 10 (31%) showed chemosensory responses only to PO2 decrease; 9 (28%) to all stimulants (PO2 decrease, PCO2 increase and pH decrease); 9 (28%) to PO2 decrease and PCO2 increase; 3 (9%) to PO2 decrease and pH decrease; only one to pH decrease. (2) The potency of the three natural stimulants in eliciting the changes in intensity of discharge showed a decreasing order as follows: PO2 decrease > PCO2 increase > pH decrease. The above results suggest that the carotid body may contain only one or a combination of 2 or 3 kinds of chemoreceptors respectively sensitive to decrease PO2 to increase PCO2 or decrease pH.