Structural analysis of substrate binding by the molecular chaperone DnaK

DnaK and other members of the 70-kilodalton heat-shock protein (hsp70) family promote protein folding, interaction, and translocation, both constitutively and in response to stress, by binding to unfolded polypeptide segments. These proteins have two functional units: a substrate-binding portion binds the polypeptide, and an adenosine triphosphatase portion facilitates substrate exchange. The crystal structure of a peptide complex with the substrate-binding unit of DnaK has now been determined at 2.0 angstroms resolution. The structure consists of a beta-sandwich subdomain followed by alpha-helical segments. The peptide is bound to DnaK in an extended conformation through a channel defined by loops from the beta sandwich. An alpha-helical domain stabilizes the complex, but does not contact the peptide directly. This domain is rotated in the molecules of a second crystal lattice, which suggests a model of conformation-dependent substrate binding that features a latch mechanism for maintaining long lifetime complexes.     

A simple technique for the long-term non-polarised and polarised culture of human fallopian tube epithelial cells

Fallopian tubes were obtained from 25 women undergoing abdominal hysterectomy. Pieces of fallopian tube mucosa were placed in culture flasks containing minimum essential medium in Earle's salts supplemented with fetal bovine serum. First passage was carried out after 7-10 days and subcultures in 4-5 days. For polarised cell culture, epithelial cells were seeded onto an extracellular matrix system. New epithelial cells were seen on day 2-3 of the primary culture and epithelial patches on day 7-10. Cells reached confluence in 4-5 days in subcultures. The cells could be subcultured for 7-11 passages with a life span of 42-60 days. Epithelial origins of the cells were confirmed by immunofluorescence staining with anti-cytokeratin antibody. Polarised cells showed a columnar pattern, microvilli on their apical surface and basally located nucleus whereas non-polarised cells were flat. It was concluded that the human fallopian tube epithelial cells can be cultured in vitro to create non-polarised and polarised cell layers by using a simple and reproducible technique and this system can be a potential model to study function of the fallopian tube.     

A surgical technique for resecting malignancies invading the facial nerve and petrous pyramid

Certain head and neck malignancies tend to invade branches of the cranial nerves and progress centripetally. The maxillary and mandibular divisions of the V (trigeminal) nerve are the most commonly invaded, but the VII (facial) nerve may harbor extensions of primary parotid malignancies and/or skin cancers originating in or near the ear. Paresis of the VII nerve signifies a serious but not entirely hopeless situation if countered with an appropriate surgical procedure. The optimal treatment of malignancies invading cranial nerves is surgical excision, which may necessitate removal of substantial portions of the cranium. Surgical pursuit of a malignancy in the facial nerve leads to a direct confrontation with the petrous pyramid of the temporal bone. Resection of the petrous pyramide is surgically feasible, but, if performed en bloc, extracts a high morbidity and mortality. An alternate technique in which the soft tissue and bones are removed disjunct is advocated. In this technique, the soft tissues are excised flush with the surface of the temporal bone. After making perforator and burr openings in the squamous temporal bone, a subtotal temporal craniectomy is completed with rongeurs. A prerequisite for success with this technique is the rotation of a large scalp flap and other local flaps to cover the exposed meninges and carotid arteries. A resume of four cases using this technique is presented. The age of the patients, the type, stage, and duration of the malignancies, and other factors which might influence the end stage results are given.     

Family structure and the mental health of children. Concurrent and longitudinal community-wide studies

This study provides a map of variations of families and some of the core relationships between types of family and the mental health of children. Family types in a poor, black urban community were defined in terms of the adults present at home. The resulting taxonomy is based on two populations: half of the community's 1964 first-grade children and families and the entire 1966 first-grade children and families. Eighty-six family types were found, falling into ten major classes. Family type was found to be strongly related over time to the child's social adaptational status (SAS) and his or her psychological well-being. The results suggest that (1) mother alone families entail the highest risk in terms of social maladaptation and psychological well-being of the child; (2) the presence of certain second adults has important ameliorative functions--mother/grandmother families being nearly as effective as mother/father families, with mother/stepfather families similar to mother alone in regard to risk; and (3) the absence of the father was less important than the aloneness of the mother in relation to risk.     

Effect of carboxylic acids on permeation of chlorpromazine through dimethyl polysiloxane membrane

The effect of carboxylic acids on the permeation of chlorpromazine was investigated through a dimethyl polysiloxane nonpolar membrane. The permeability of the diffusate, at pH 5.8, increases considerably in the presence of carboxylic acids or phosphate, probably due to an ion-pair formation between the relative anions and chlorpromazine.     

Diurnal rhythms in pineal N-acetyltransferase and hippocampal norepinephrine: effects of water deprivation, blinding and hypothalamic lesions

The pineal gland in the rat exhibits a diurnal rhythm in activity of the enzyme serotonin N-acetyltransferase (N-AT) with peak values during the dark period of a diurnal lighting schedule approximately 100-fold those during the light period. After blinding the rhythm becomes free-running. It is abolished by partial hypothalamic deafferentation with a knife cut made caudal to the optic chiasm. Water deprivation for 23 h daily has no effect on the pineal rhythm in either intact, blinded or deafferented animals. In contrast to this, there is a diurnal rhythm in hippocampal formation in norepinephrine content which can be entrained by a water deprivation schedule in both intact and blinded animals. These observations indicate that in the same animals 1 diurnal rhythm may remain entrained to the light-dark cycle while another rhythm is entrained to a secondary synchronizer, the water deprivation schedule.     

Mechanism of production of intestinal secretion by negative luminal pressure

The relationships between luminal hydrostatic pressure and fluid transport by dog jejunum in vivo studied as a sheet in the Wells clamp were compared with quantitative predictions from a model proposed for the mechanism of the secretion produced by elevated venous pressure. According to the model, the secretion produced by increasing venous pressure and the secretion produced by negative luminal pressure are both passive filtrates contingent on a transepithelial pressure of a few centimeters of H2O. We consider that the agreement between the observed and predicted responses to luminal pressure provides strong support for the model. In particular, a) the observations displayed a predicted gross asymmetry in rates of fluid transfer with isotonic fluids depending on whether the luminal pressure was positive or negative; b) the observed magnitude of the negative luminal pressure required for the onset of secretion agreed with predictions; and c) the secretion contained significant amounts of protein at about 25% of the plasma concentration.     

Cardiopulmonary effects of intermittent mandatory ventilation

IMV is a combination of spontaneous and mechanical ventilation. For numerous reasons, IMV is potentially more advantageous than conventional techniques. By maintaining spontaneous breathing, mechanical augmentation can be titrated to adjust alveolar minute ventilation levels to normal, thereby decreasing the incidence of respiratory alkalemia. There are major differences between the cardiopulmonary effects of IMV and conventional mechanical ventilation. Spontaneous inspiration decreases Ppl and results in better distribution of inspired gas, a better V/Q, and less physiological dead space. In addition, transmural filling pressures, venous return, and cardiac output are more normal than during conventional mechanical ventilation. Maintenance of spontaneous ventilation lowers mean Paw and pulmonary vascular resistance. If venous admixture occurs, it can be minimized by titrating PEEP. Thus, more effective therapy for hypoxemia is possible. If spontaneous breathing is to persist and be efective, work-of-breathing must be minimized. This can be accomplished best when a continuous flow of gas provides optimal CPAP to maintain FRC and to minimize the effects of decreased compliance without depressing cardiac function.