Mariner 9 television reconnaissance of Mars and its satellites: preliminary results.

At orbit insertion on 14 November 1971 the Martian surface was largely obscured by a dust haze with an extinction optical depth that ranged from near unity in the south polar region to probably greater than 2 over most of the planet. The only features clearly visible were the south polar cap, one dark, spot in Nix Olympica, and three dark spots in the Tharsis region. During the third week the atmosphere began to clear and surface visibility improved, but contrasts remained a fraction of their normal value. Each of the dark spots that apparently protrude through most of the dust-filled atmosphere has a crater or crater complex in its center. The craters are rimless and have featureless floors that, in the crater complexes, are at different levels. The largest crater within the southernmost spot is approximately 100 kilometers wide. The craters apparently were formed by subsidence and resemble terrestrial calderas. The south polar cap has a regular margin, suggsting very flat topography. Two craters outside the cap have frost on their floors; an apparent crater rim within the cap is frost free, indicating preferentia loss of frost from elevated ground. If this is so then the curvilinear streaks, which were frost covered in 1969 and are now clear of frost, may be low-relief ridges. Closeup pictures of Phobos and Deimos show that Phobos is about 25 +/-5 by 21 +/-1 kilometers and Deimos is about 13.5 +/- 2 by 12.0 +/-0.5 kilometers. Both have irregular shapes and are highly cratered, with some craters showing raised rims. The satellites are dark objects with geometric albedos of 0.05.

The human orphan nuclear receptor PXR is activated by compounds that regulate CYP3A4 gene expression and cause drug interactions.

The cytochrome P-450 monooxygenase 3A4 (CYP3A4) is responsible for the oxidative metabolism of a wide variety of xenobiotics including an estimated 60% of all clinically used drugs. Although expression of the CYP3A4 gene is known to be induced in response to a variety of compounds, the mechanism underlying this induction, which represents a basis for drug interactions in patients, has remained unclear. We report the identification of a human (h) orphan nuclear receptor, termed the pregnane X receptor (PXR), that binds to a response element in the CYP3A4 promoter and is activated by a range of drugs known to induce CYP3A4 expression. Comparison of hPXR with the recently cloned mouse PXR reveals marked differences in their activation by certain drugs, which may account in part for the species-specific effects of compounds on CYP3A gene expression. These findings provide a molecular explanation for the ability of disparate chemicals to induce CYP3A4 levels and, furthermore, provide a basis for developing in vitro assays to aid in predicting whether drugs will interact in humans.

Structure of the Tie2 RTK domain: self-inhibition by the nucleotide binding loop, activation loop, and C-terminal tail.

BACKGROUND: Angiogenesis, the formation of new vessels from the existing vasculature, is a critical process during early development as well as in a number of disease processes. Tie2 (also known as Tek) is an endothelium-specific receptor tyrosine kinase involved in both angiogenesis and vasculature maintenance. RESULTS: We have determined the crystal structure of the Tie2 kinase domain to 2.2 A resolution. The structure contains the catalytic core, the kinase insert domain (KID), and the C-terminal tail. The overall fold is similar to that observed in other serine/threonine and tyrosine kinase structures; however, several unique features distinguish the Tie2 structure from those of other kinases. The Tie2 nucleotide binding loop is in an inhibitory conformation, which is not seen in other kinase structures, while its activation loop adopts an "activated-like" conformation in the absence of phosphorylation. Tyr-897, located in the N-terminal domain, may negatively regulate the activity of Tie2 by preventing dimerization of the kinase domains or by recruiting phosphatases when it is phosphorylated. CONCLUSION: Regulation of the kinase activity of Tie2 is a complex process. Conformational changes in the nucleotide binding loop, activation loop, C helix, and the C-terminal tail are required for ATP and substrate binding.

Expression of wild-type estrogen receptor beta and variant isoforms in human breast cancer.

It has been shown in previous studies that a variety of estrogen receptor (ER) beta mRNA transcripts are expressed in human breast cancer cell lines and tumors. To complement the RNA expression studies, we have developed ER-beta-specific antibodies to characterize ER-beta protein expression in breast cancer cell lines and tumors. Monoclonal antibodies were made against a peptide representing the first 18 amino acids of the longest ER-beta open reading frame reported to date, and polyclonal antibodies were made against a peptide within the ER-beta B domain. By Western blot analysis, we show that ER-beta protein is expressed in all cancer cell lines tested and in three of five breast tumor samples. The breast cancer cell lines showed variation in the size of the expressed ER-beta protein. The longest form detected was consistent with the 530-amino acid, full-length ER-beta sequence. Shorter ER-beta isoforms were detected in the ER-alpha-negative MDA-MB-231 and MDA-MB-435 breast cancer cell lines, likely corresponding to previously described COOH-terminal RNA variant isoforms.

Applications of S-layers.

The wealth of information existing on the general principle of S-layers has revealed a broad application potential. The most relevant features exploited in applied S-layer research are: (i) pores passing through S-layers show identical size and morphology and are in the range of ultrafiltration membranes; (ii) functional groups on the surface and in the pores are aligned in well-defined positions and orientations and accessible for binding functional molecules in very precise fashion; (iii) isolated S-layer subunits from many organisms are capable of recrystallizing as closed monolayers onto solid supports at the air-water interface, on lipid monolayers or onto the surface of liposomes. Particularly their repetitive physicochemical properties down to the subnanometer scale make S-layers unique structures for functionalization of surfaces and interfaces down to the ultimate resolution limit. The following review focuses on selected applications in biotechnology, diagnostics, vaccine development, biomimetic membranes, supramolecular engineering and nanotechnology. Despite progress in the characterization of S-layers and the exploitation of S-layers for the applications described in this chapter, it is clear that the field lags behind others (e.g. enzyme engineering) in applying recent advances in protein engineering. Genetic modification and targeted chemical modification would allow several possibilities including the manipulation of pore permeation properties, the introduction of switches to open and close the pores, and the covalent attachment to surfaces or other macromolecules through defined sites on the S-layer protein. The application of protein engineering to S-layers will require the development of straightforward expression systems, the development of simple assays for assembly and function that are suitable for the rapid screening of numerous mutants and the acquisition of structural information at atomic resolution. Attention should be given to these areas in the coming years.

The pregnane X receptor: a promiscuous xenobiotic receptor that has diverged during evolution.

Transcription of genes encoding cytochrome P450 3A (CYP3A) monooxygenases is induced by a variety of xenobiotics and natural steroids. There are marked differences in the compounds that induce CYP3A gene expression between species. Recently, the mouse and human pregnane X receptor (PXR) were shown to be activated by compounds that induce CYP3A expression. However, most studies of CYP3A regulation have been performed using rabbit and rat hepatocytes. Here, we report the cloning and characterization of PXR from these two species. PXR is remarkably divergent between species, with the rabbit, rat, and human receptors sharing only approximately 80% amino acid identity in their ligand-binding domains. This sequence divergence is reflected by marked pharmacological differences in PXR activation profiles. For example, the macrolide antibiotic rifampicin, the antidiabetic drug troglitazone, and the hypocholesterolemic drug SR12813 are efficacious activators of the human and rabbit PXR but have little activity on the rat and mouse PXR. Conversely, pregnane 16alpha-carbonitrile is a more potent activator of the rat and mouse PXR than the human and rabbit receptor. The activities of xenobiotics in PXR activation assays correlate well with their ability to induce CYP3A expression in primary hepatocytes. Through the use of a novel scintillation proximity binding assay, we demonstrate that many of the compounds that induce CYP3A expression bind directly to human PXR. These data establish PXR as a promiscuous xenobiotic receptor that has diverged during evolution.

Inguinal hernia repair: results using an open preperitoneal approach.

BACKGROUND: Laparoscopic surgical approaches to the repair of inguinal hernias have shown the advantages of placing mesh in the preperitoneal space. Despite those advantages, laparoscopic hernia repairs have been associated with increased cost, longer operating times, and advanced laparoscopic skills. An open preperitoneal approach has the benefit of mesh in the preperitoneal position without the disadvantages of a laparoscopic procedure. METHODS: We present our experience with the use of an open preperitoneal mesh repair (KugelMesh, Bard, Inc.). The study was conducted in a prospective fashion from January 1998 through October 2001. 1072 hernias were repaired in two community hospitals by three general surgeons. Patients with recurrent hernias were excluded if the initial repair was from a preperitoneal approach. Operative time, cost, post-operative pain, and complications were analyzed. RESULTS: Recurrences occurred in five patients (0.47%) during a mean follow-up time of 23 months (range: 2-47). The average operating time was 32.4 min (range: 16-62). Post-operative narcotic pain medication usage averaged 5.8 pills (range: 0-26) per repair. Average surgical charges were less for the open preperitoneal approach ($2253) than for laparoscopic repairs ($4826). CONCLUSIONS: The open preperitoneal hernia repair using the Kugel mesh offers many advantages. It is inexpensive, has a low recurrence rate, and allows the surgeon to cover all potential defects with one piece of mesh. Postoperative recovery is short and postoperative pain is minimal.

Opiate addiction and suicidal behaviors.

The authors tested the hypothesis that people addicted to opiates manifest more psychopathology in areas that have been identified as predictors for high risk of suicidal behavior than do normal control subjects. They gave 278 patients in a methadone maintenance program and 207 normal control subjects the index of Potential Suicide (IPS), a scale designed to assess suicidal risk. Using discriminate function analysis, the authors found that 87% of 100 of the methadone patients and 98% of 100 of the normal control subjects were correctly identified on the basis of the IPS data.

Magnetic resonance findings in REM sleep behavior disorder.

Rapid eye movement (REM) sleep behavior disorder is characterized by bizarre acts during nocturnal sleep that may lead to physical injuries. Dream content suggests that motor overactivity is an attempted dream enactment and polygraphic studies reveal REM stage without atonia, an alteration of REM sleep generation that facilitates excessive motor activity. In 6 patients with REM sleep behavior disorder. MRI of the brain showed multifocal signal intensity lesions suggestive of lacunar infarcts in periventricular regions (5 patients) and in dorsal pontomesencephalic areas (3 patients). REM sleep behavior disorder may be the result of injury to the midrostral tegmentum nuclei, the tegmentoreticular tracts, or both. This condition is easily controlled with clonazepam.

Pharmacophore analysis of the nuclear oxysterol receptor LXRalpha.

A cell-free assay was developed for the orphan nuclear receptor LXRalpha that measures the ligand-dependent recruitment of a peptide from the steroid receptor coactivator 1 (SRC1) to the nuclear receptor. Using this ligand-sensing assay (LiSA), the structural requirements for activation of the receptor by oxysterols and related compounds were studied. The minimal pharmacophore for receptor activation was shown to be a sterol with a hydrogen bond acceptor at C24. 24(S),25-Epoxycholesterol (1), which meets this criterion, is among the most efficacious of the oxysterols and is an attractive candidate as the LXRalpha natural hormone. Cholenic acid dimethylamide (14) showed increased efficacy compared to 1, whereas the unnatural oxysterol 22(S)-hydroxycholesterol (4) was shown to be an antagonist of 1 in the LiSA. The structural requirements for SRC1 recruitment in the LiSA correlated with the transcriptional activity of compounds in a cell-based reporter assay employing LXRalpha-GAL4 chimeric receptors. Site-directed mutagenesis identified Trp(443) as an amino acid critical for activation of LXRalpha by oxysterol ligands. This information was combined with the structure-activity relationship developed from the LiSA to develop a 3D homology model of LXRalpha. This model may aid the design of synthetic drugs targeted at this transcriptional regulator of cholesterol homeostasis.

Relationship between sleep spindles and hypersomnia.

Sleep spindles (SS) and K complexes constitute the physiological markers of stage 2 sleep. Because sleep allows a spontaneous thalamic manifestation in the form of SS, one could hypothesize that there is some kind of relationship between SS and the complaint of hypersomnia. To investigate this possible relationship we compared nonhypersomnolent subjects with hypersomnolent patients who carried a diagnosis of narcolepsy or idiopathic hypersomnia. SS were counted in well-defined nocturnal stage 2 sleep segments, and the average SS density (number of SS in stage 2/minute stage 2) was tabulated for the entire night. Agreement between two independent scores was higher than 95%. The results show that the average SS density is higher in both cerebral hemispheres in the hypersomnolent group, especially in the idiopathic hypersomnia patients. At the beginning and at the end of the nocturnal sleep time, SS density is increased in this group compared with the normal one. These findings support the complaint of hypersomnia, mainly in idiopathic hypersomnia patients. This is in agreement with the notion that SS are generated by thalamic structures that serve a gatekeeping function during nonrapid eye movement sleep, and further suggests that their relative abundance expresses the power of that control.

The effects of androgen on the transcription of specific genes in guinea pig seminal vesicle epithelium.

Steroid hormones have been shown to have highly differential effects on the expression of abundant cell-specific protein genes in a multitude of model tissues. In rat seminal vesicle, for example, DNA clones representing two major secretory protein genes have been used to show that both of the genes are differentially regulated by androgen. In this paper, we have examined the effects of androgen on the transcription of two major secretory protein genes in guinea pig seminal vesicle epithelium. Nuclear run-off experiments were used to show that castration of the adult resulted in a 3-fold decrease in total transcription activity. Surprisingly, the decrease in total transcriptional activity was not reflected in a differential decrease in the transcriptional activity of the two major secretory protein genes. When the effects of castration on the transcriptional activity of the major secretory protein genes were compared to the effects on other genes, it was found that the transcriptional activity of each gene examined was decreased by the same magnitude as the major secretory protein genes. Similarly, the transcriptional activity of every gene examined increased by the same magnitude as the major secretory protein genes during hormone repletion of the castrated adult. Thus, in contrast to the differential effects of steroids on the transcription of abundant cell-specific proteins in many other steroid-dependent tissues, the transcription of major secretory proteins in guinea pig seminal vesicle epithelium appears to be regulated in parallel with many other genes. The generalized effects of androgen on transcriptional activity could account for the generalized effects of androgen on seminal vesicle epithelial cell structure and function.

Recognition and treatment of depression in a family medicine practice.

To test the hypothesis that depression is significantly underdiagnosed in general medical settings, the Zung Self-Rating Depression Scale was administered to 1,086 family medicine outpatients seen during a 12-month period before their initial medical examination. The effects of such screening on clinical recognition and treatment of depression were examined. Of the 1,086 patients, 143 (13.2%) were symptomatically depressed. These patients were randomized into two groups: 102 were identified as clinically depressed to their physician, and the remaining 41 were not (control group). Physicians diagnosed depression in 15% of the control group and in 68% of the identified group. At 4-week follow-up, 64% of the identified patients who were treated with maprotiline (16 of 25) showed improvement; only 28% of the identified patients who were electively not treated improved. Improvement occurred in only 18% of the control group. It appears that the diagnosis of depression is not ordinarily made in family medicine outpatient settings and that self-rating depression scales are useful diagnostic aids, whose regular use is indicated by the high prevalence of depression in general medical populations.

Evolution of a geriatric evaluation clinic.

Multidisciplinary geriatric evaluation units are being established in hospitals and, to a lesser extent, as outpatient clinics. This paper presents results of a chart audit of 332 patients seen from 1978 to 1982 at a university based geriatric outpatient evaluation unit. The relationships of clinic staffing to types of referrals and other aspects of clinic operation are investigated. During the time under study, the proportion of patients whose problems were medical rather than psychiatric or social increased. Presenting problems varied by referral source. Family and self referrals were most likely to identify a medical presenting problem, while physician and community agency referrals were most likely to identify a psychiatric problem. Patients frequently received diagnoses in areas of function other than those identified as the presenting problems. For example, medical clinicians made at least one psychiatric diagnoses for 86 per cent of patients they evaluated. Important questions about the potential contribution of such geriatrics clinics to health care of the elderly remain to be answered.

Assessment of elderly patients at home: a report of fifty cases.

Fifty elderly patients were assessed at home by physicians trained in geriatric medicine. The patients had been referred by physicians, relatives, or community services. Reasons for referral usually were nonspecific and involved either a request for overall assessment or recommendations about placement. Assessment included medical, psychiatric, social and functional components. Mental impairment and impairment in the activities of daily living were common. New medical or psychiatric diagnoses were contributed in 36 instances. The most frequent previously unnoticed pathologic disorders were mental impairment and depression. Assessment resulted in specialists' advice concerning such matters as the medication program, placement, increased support, further diagnostic work and, in two instances, prompt admission to a hospital.

Identifying substrates for endothelium-specific Tie-2 receptor tyrosine kinase from phage-displayed peptide libraries for high throughput screening.

The peptide substrate specificity of Tie-2 was probed using the phage display method in order to identify efficient substrate for high throughput screening. Two random peptide libraries, pGWX3YX4 and pGWX4YX4, were constructed, in which all twenty amino acid residues were represented at the X positions flanking the fixed tyrosine residue Y. A fusion protein of GST and the catalytic domain of human Tie-2 was used to perform the phage phosphorylation. The phosphorylated phage particles were enriched by panning over immobilized anti-phosphotyrosine antibody pY20 for a total of 5 rounds. Four phage clones (3T61, 3T68, C1-90 and D1-15) that express a peptide sequence that can be phosphorylated by the recombinant catalytic domain of human Tie-2 were identified. Synthetic peptides made according to the sequences of the 4 selected clones from the two libraries, which had widely different sequences, were active substrates of Tie-2. Kinetic analysis revealed that D1-15 had the best catalytic efficiency with a k(cat)/K(m) of 5.9x10(4) M(-1) s(-1). Three high throughput screening assay formats, dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA), radioactive plate binding (RPB) and time-resolved fluorescent resonance energy transfer (TR-FRET) were developed to assess the suitability of these phage display selected peptides in screening Tie-2 inhibitors. Three out of four peptides were functional in the DELFIA assay and D1-15 was functional in the TR-FRET assay.

Thoracoscopic excision of mediastinal parathyroid adenomas: a report of two cases and review of the literature.

BACKGROUND: Most abnormal parathyroid glands can be removed through a standard cervical incision; even those in the superior mediastinum. Those located in certain areas of the mediastinum, for example posteriorly or in the aortopulmonic window, historically have required excision through a median sternotomy or thoracotomy. Angioablation is a nonsurgical alternative to management of these lesions. STUDY DESIGN: We present two case reports of mediastinal parathyroid adenomas that were excised thoracoscopically, and review the literature regarding the management of mediastinal parathyroid adenomas. RESULTS: Both patients who underwent precise localization and thoracoscopic excision of their mediastinal parathyroid adenomas had resolution of their hypercalcemia with minimal associated morbidity and shortened recovery periods. CONCLUSIONS: We suggest that thoracoscopic excision of mediastinal parathyroid adenomas is the better means of controlling hypercalcemia secondary to parathyroid adenoma in those patients considered for either median sternotomy, thoracotomy or angiographic ablation where the exact location of the lesion can be established preoperatively.

Use of the nuclear receptor PXR to predict drug interactions.

We recently cloned the human, rabbit, rat, and mouse orthologs of a novel member of the steroid/retinoid/thyroid hormone receptor family, which we have named the Pregnane X Receptor (PXRs). The discovery and characterization of PXR has led to an increased understanding of the molecular basis of many drug-drug interactions as well as a better understanding of xenobiotic metabolism in general. The key insights into PXR action was the finding that this nuclear receptor is linked to regulation of the cytochrome P450 3A monooxygenase (CYP3A) genes. Several lines of evidence indicate that PXR mediates the induction of CYP3A gene transcription. First, PXR is selectively expressed in the liver and intestine, the same tissues in which CYP3A gene expression is induced. Second, PXR binds as a heterodimer with the retinoid X receptor (RXR) to xenobiotic response elements that have been identified in CYP3A gene promoters. Third, PXR is activated by the remarkable array of compounds that are known to induce CYP3A gene transcription. And finally, PXRs from different species are differentially activated by certain compounds such as rifampicin and pregnenolone 16alpha-carbonitrile (PCN) in a manner that correlates with species-specific induction of CYP3A gene expression. We are now employing high throughput PXR activation and binding assays to identify drug candidates that induce CYP3A gene expression so that these compounds can be removed from the drug development process.

Malignant pneumomediastinum: successful tube mediastinostomy in the neonate.

Five critically ill neonates underwent tube mediastinostomy in the neonatal intensive care nursery for tension pneumomediastinum. All of the neonates showed immediate clinical improvement, with a decrease in peak airway pressure and elevation of arterial oxygen pressure levels. There were no complications directly attributable to the procedure. We believe that pneumomediastinum in association with severe hypoxia, metabolic acidosis, and high ventilation pressures indicates clinically significant tension in the mediastinum. This results in a decrease in systemic blood pressure and pulmonary venous return that is not amenable to conservative management. Needle aspiration is inadequate because of the dynamic nature of the air leak. Tube decompression of the mediastinum is the treatment of choice in these circumstances.

Vertical ring gastroplasty for morbid obesity. Five year experience with 1,463 patients.

Vertical gastroplasty is a safer and simpler operation than gastric bypass for morbid obesity, but to produce permanent long-term weight loss, a permanent support is needed for the outlet of the newly created gastric pouch in vertical gastroplasty. The most important finding in this study of 1,463 patients with vertical gastroplasties was the development of a safe and practical use of a Silastic ring to permanently support the outlet. Using an uncovered Silastic ring, erosion of the ring has not been observed and long-term weight loss of 36.7 percent of original weight or 63.4 percent of excess weight has been obtained, which is equivalent to that reported for gastric bypass or other gastric reduction procedures. In addition, lower mortality and lower morbidity rates have been achieved than those reported in studies using other gastric reduction methods. Silastic ring vertical gastroplasty with an uncovered ring is the safest and most effective of the gastric reduction procedures in our experience.