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Succinate production in dual-phase Escherichia coli fermentations depends on the time of transition from aerobic to anaerobic conditions.

We examined succinic acid production in Escherichia coli AFP111 using dual-phase fermentations, which comprise an initial aerobic growth phase followed by an anaerobic production phase. AFP111 has mutations in the pfl, ldhA, and ptsG genes, and we additionally transformed this strain with the pyc gene (AFP111/pTrc99A-pyc) to provide metabolic flexibility at the pyruvate node. Aerobic fermentations with these two strains were completed to catalog physiological states during aerobic growth that might influence succinate generation in the anaerobic phase. Activities of six key enzymes were also determined for these aerobic fermentations. From these results, six transition times based on physiological states were selected for studying dual-phase fermentations. The final succinate yield and productivity depend greatly on the physiological state of the cells at the time of transition. Using the best transition time, fermentations achieved a final succinic acid concentration of 99.2 g/l with an overall yield of 110% and productivity of 1.3 g/l h.

Aerobiosis↗

Type II keratin cDNAs from the rainbow trout: implications for keratin evolution.

From a teleost fish, the rainbow trout Oncorhynchus mykiss, we have cloned and sequenced cDNAs encoding five different type II keratins. The corresponding protein spots, as separated by 2D-PAGE of trout cytoskeletal preparations, have been identified by peptide mass mapping using MALDI mass spectrometry. Three of the sequenced keratins are expressed in the epidermis (subtype IIe), and two in simple epithelia and mesenchymal cells (subtype IIs). The IIs keratins are both orthologs of human K8. This leaves unsequenced only the trace component S3 of the biochemically established trout keratin catalog. A phylogenetic tree has been constructed from a multiple alignment of the rod domains of the new keratin sequences together with type II sequences from other vertebrates such as shark, zebrafish, and human; lamprey K8 (recently sequenced in our laboratory) has been used as outgroup. This tree suggests, in a highly bootstrap-supported manner, that the teleost IIe keratins diversified independently from the mammalian IIe keratins. In contrast, all the species investigated express K8-like keratins, suggesting that the different IIe branches evolved from K8-like progenitors. The tree also indicates that the published zebrafish sequences represent IIe keratins and that the biochemically identified K8 ortholog in zebrafish has not yet been sequenced.

Amino Acid Sequence↗

Migraine prophylactic drugs: proof of efficacy, utilization and cost.

OBJECTIVES: In order to understand the pattern of utilization of migraine prophylactic drugs by US physicians, we reviewed the scientific rigor of published trials of anti-migraine medications, assessed their cost, and tested the correlation, if any, between utilization, scientific rigor and cost. MATERIALS AND METHODS: Scientific rigor of published reports. We identified all placebo-controlled, randomized, double-blind trials of migraine prophylactic agents through Medline search, major Headache textbooks and proceedings of major scientific meetings where headache-related topics are discussed. We excluded trials that did not include placebo treatment during the active phase of the study. The trials were reviewed and rated for scientific rigor using a 5-point scale (scientific score [ss]; 1 = low, 5 = good), blinded to the physicians' utilization data and cost of the drugs. Studies that did not show benefit of the active drug over placebo were scored -1 to -5, thus allowing for the reverse logic of negative studies. US physicians utilization. Neurologists and primary care physicians (PCP) completed phone-mail-phone questionnaires which inquired about first and second choices of migraine prophylaxis. These choices were averaged to obtain a weighted average percent usage of each drug. Cost. The average wholesale price (AWP) of each drug was obtained from data published by Adelman and Von Seggern, and from the Amerisource (7/9/96) catalog. STATISTICAL ANALYSIS: Spearman's correlation coefficient was used to assess the relationship between the average ss, physician use, and cost of each drug. RESULTS: Propranolol (ss = 1.44), amitriptyline (ss = 2.33) and verapamil (ss = 1.00) were the three preferred migraine prophylactic drugs by both neurologists and PCPs. Approximately 10% of neurologists said that divalproex (ss = 3.75) would be their first or second choice. The selective serotonin reuptake blockers were favored by 13.21% of PCPs. All other prophylactic drugs were felt to be first or second line of treatment by less than 10% of either neurologists or PCPs. Except for one study (ss = 1) that showed that propranolol reduced the migraine frequency by 76% over placebo, trials of the three most preferred medications failed to demonstrate that the active drug is > 50% better than placebo, i.e. the difference in headache frequency when on placebo vs active drug is > 50%. Of the drugs available in the United States, flurbiprofen and metoprolol achieved the best ss (5.00 and 4.33, respectively) but their efficacy over placebo (23% and 14-33%, respectively) and cost ($67.2 and $65.6) were unfavorable. Neurologists and PCPs chose migraine prophylaxis on the basis of scientific merit (r = 0.644, p = 0.018; r = 0.576, p = 0.05, respectively) but not cost (r = -0.254, p = 0.45; r = -0.255, p = 0.455). CONCLUSION: The three most commonly chosen migraine prophylactic agents have not been shown irrefutably to prevent migraine. Furthermore, their benefit, if any, does not exceed 50% over placebo. The well-conducted recent trials that demonstrated the efficacy of divalproex in migraine prevention are steps in the right direction of finding the "ideal migraine preventative agent". Until that drug is discovered, it is difficult to argue that one migraine prophylactic medication is superior to another and accordingly should be used as a first line of treatment.

Adult↗

Severe palmo-plantar hyperkeratosis in Dowling-Meara epidermolysis bullosa simplex caused by a mutation in the keratin 14 gene (KRT14).

Mutant keratins 5 or 14 are implicated in the etiology of epidermolysis bullosa simplex (EBS). The catalog of mutations has established certain patterns of mutation clusters from which it may be possible, along with associated biochemical data, to predict phenotypic severity. It is becoming apparent that some of these assumptions may now require modification. We report a mutation in the gene encoding keratin 14 (KRT14) that changes the predicted amino acid at position 119, at the start of the helix initiation motif, from methionine to threonine (K14 M119T) in a patient with an EBS Dowling-Meara phenotype with severe palmo-plantar hyperkeratosis. This demonstrates that the three major types of EBS can arise from missense mutations in the same codon. The findings suggest that the specific nature of the missense mutation, in the context of the protein sequence, can contribute far more to the clinical severity than previously thought. The different EBS subtypes should be viewed as gradations of clinical severity rather than distinct genetic diseases.

Adult↗

The hemodialysis catheter conundrum: hate living with them, but can't live without them.

BACKGROUND: Hemodialysis requires reliable recurrent access to the circulation. On a chronic basis, this has been best provided by the use of arteriovenous fistulae and arteriovenous grafts. In recent years, hemodialysis catheters have come to play an increasingly important role in the delivery of hemodialysis. The use of both temporary as well as cuffed hemodialysis catheters has emerged as a significant boon for both patients and practicing nephrologists. The complications, however, associated with each of these hemodialysis catheters, both in terms of anatomic, thrombotic, and infectious issues, have emerged as a major problem with their continued use. This significant morbidity and complication rate has forced many nephrologists to face a basic conundrum: they have come to hate having to deal with the problems inherent in catheter usage, but the enormous utility of these devices have forced physicians to accept the fact they cannot live without them in their current practice. METHODS: We used a comprehensive literature review to describe the types, use and dilemmas of hemodialysis catheters. RESULTS: This article provides a comprehensive review of both the benefits inherent with the use of these hemodialysis catheters while cataloging their complications and offering some possible solutions. CONCLUSION: Hemodialysis vascular access catheters are essential in the maintenance of hemodialysis vascular access. However, they have a significant infectious, thrombotic, anatomic complication rate that are detailed with proposed problem-solving guidelines.

Catheters, Indwelling↗

Silicone versus nonsilicone gel dressings: a controlled trial.

BACKGROUND: Silicone gel dressings decrease scar volume and soften hypertrophic tissue, allowing it to be more easily controlled by other methods. Although silicone does not appear to be an essential component of the treatment, nonsilicone dressings have been reported to cause no change in physical parameters during a 2-month treatment period. OBJECTIVE: To compare silicone and nonsilicone gel dressings in the treatment of keloids and hypertrophic scars, including a control group, and to evaluate the effectiveness of these treatments using two new assessment techniques. METHODS: Patients were randomly chosen to receive silicone or nonsilicone gel dressings in a 4.5-month controlled prospective study. Scar size, induration, and symptoms were evaluated before and after the treatment. Scar color was visually measured using a color palette catalog, and a new device was developed to measure intracicatricial pressure. RESULTS: All of the measured parameters were significantly reduced in both silicone- and nonsilicone-treated groups, as compared to the control, with no significant differences between them. CONCLUSION: Silicone and nonsilicone gel dressings are equally effective in the treatment of keloids and hypertrophic scars.

Adolescent↗

Evaluation of monoclonal antibody MIB-1 in the mammary epithelium adjacent to fibroadenomas in premenopausal women treated with tamoxifen.

The purpose of this study was to study the monoclonal antibody MIB-1 in the normal breast epithelium adjacent to a fibroadenoma in women in the luteal phase of the menstrual cycle who were treated with tamoxifen at doses of 10 and 20 mg for 22 days. The proliferative activity of the mammary epithelium adjacent to the fibroadenoma was studied by immunohistochemistry on the basis of the monoclonal antibody MIB-1 (Immunotech, catalog No. 0505, lot 001). The study was randomized and double blind and was conducted on 44 women with fibroadenomas divided into three groups: A (n=16, placebo), B (n=15, tamoxifen, 10 mg), and C (n=13, tamoxifen, 20 mg). Tamoxifen was administered for 22 days starting on the 2nd day of the menstrual cycle, and a biopsy was taken on the 23rd day. Serum estradiol, progesterone, sex hormone binding globulin, follicle-stimulating hormone, luteinizing hormone, and prolactin were measured before treatment (21st and 24th day of the previous menstrual cycle) and on the day of the biopsy. The mean percentage of stained nuclei per 1,000 cells was 9.2 in group A, 4.5 in group B, and 3.2 in group C. The Fisher's test revealed that tamoxifen significantly reduced MIB-1 at doses of 10 and 20 mg compared with the placebo group (p < 0.0001), with no significant differences between doses in terms of proliferative activity (p=0.21). Groups B and C presented a significant increase in progesterone (p=0.038), estradiol (p < 0.001), and sex hormone binding globulin (p=0.001) levels. Elevation of serum follicle-stimulating hormone concentration (p=0.0045) and a fall in prolactin levels (p=0.0055) were observed. We conclude that tamoxifen significantly reduced the proliferative activity of the mammary epithelium at the doses of 10 and 20 mg/day.

Adolescent↗

Ambulatory morning report: can it prepare residents for the American Board of Internal Medicine Examination?

We assessed the ability of a novel ambulatory morning report format to expose internal medicine residents to the breadth of topics covered by the American Board of Internal Medicine (ABIM) exam. Cases were selected by the Ambulatory Assistant Chief Residents and recorded in a logbook to limit duplication. We conducted a retrospective review of 406 cases discussed from July 1998 to July 2000 and cataloged each according to the primary content area. The percentage of cases in each area accurately reflected that covered by the ABIM exam, with little redundancy or over-selection of esoteric diseases. Our data suggest that a general medicine clinic is capable of exposing house staff to the wide breadth of internal medicine topics previously thought to be unique to subspecialty clinics.

Ambulatory Care↗

Keratin 1 expression in endothelia and mesenchymal tumors: an immunohistochemical analysis of normal and neoplastic tissues.

Keratin polypeptides of the nonhair type, numbered 1 through 20 in the Moll catalog, are selectively expressed in normal and neoplastic tissues. Keratin 1 (K1), the highest-molecular-weight keratin (67 kd), was generally considered specific for keratinizing squamous epithelia. However, recent studies have shown that it is an integral component of the multiprotein kininogen receptor of endothelial cells. A library of formalin-fixed, paraffin-embedded tissue samples was evaluated immunohistochemically (avidin-biotin peroxidase complex method) for K1 expression using a specific monoclonal antibody (Novocastra clone 34betaB4). The study group included a wide variety of normal tissues and 541 tumors of epithelial or mesenchymal derivation. The specificity of the antibody to K1 was verified in normal epithelial tissues, where the staining was essentially limited to the epidermis and Hassal corpuscles of the thymus and focally to other squamous epithelia. Among carcinomas, it was essentially limited to keratinizing squamous carcinomas. It was also regularly found in endothelial cells of normal capillaries, veins, and arteries. Capillary, cavernous, and venous hemangiomas often had endothelia with K1 positivity. Among the malignant vascular tumors, epithelioid hemangioendotheliomas were consistently positive (8 of 8). However, angiosarcomas had more variable expression (59 of 81 were positive), with well-differentiated tumors generally having greater reactivity than poorly differentiated examples. Mesenchymal tumors with K1 expression included schwannomas (10 of 16), epithelioid sarcomas (26 of 37), and synovial sarcomas (19 of 68). In the last 2 tumor types, K1 reactivity was detected in both epithelioid and spindled neoplastic populations. In addition to its specificity for keratinizing squamous epithelia, K1 can be immunohistochemically detected in normal vascular endothelial cells and a spectrum of vascular tumors. However, its expression in poorly differentiated vascular tumors is variable, suggesting that this marker is poorly conserved in highly transformed endothelia. The unexpected K1 immunoreactivity in nonvascular soft tissue tumors, such as synovial sarcoma, epithelioid sarcoma, and schwannomas, requires further study.

Adult↗

Variation in the human genome and the inherited basis of common disease.

The availability of a reference human genome sequence-an increasingly dense catalog-knowledge of common genetic variation, and new developments in technology present an unprecedented opportunity to systematically explore the genetic basis of complex human diseases such as cancer. An understanding of the common mutations that can cause distinct human cancers will be critical for identifying new targets for drug discovery, patient stratification for clinical trials, and analysis of drug response data to delineate classes of patients that respond to therapy. The genome structure of cancer can be investigated in several ways. Germline mutations can be investigated in large-scale, case-control, or family studies. Somatic alternations can be identified using state-of-the-art genomic technologies such as high-density oligonucleotide arrays and targeted resequencing. Combined, these approaches will lead to a better understanding of the cancer genome.

Gene Expression Profiling↗

Electrosurgical methods for arthroscopic meniscectomy: A review of the literature.

PURPOSE: To outline the development of electrosurgical (radiofrequency) devices, explain the basic principles, and review the current orthopaedic literature regarding the application of electrosurgery to arthroscopic partial meniscectomy. MATERIALS AND METHODS: The history and principles of electrosurgery were obtained from various pertinent texts and journal articles. A literature search was performed using MEDLINE; reviewed articles consisted of articles in the English language cataloged between 1966 and January 1999. RESULTS: The history and principles of electrosurgery are reviewed. The articles pertaining to arthroscopic electrosurgical meniscectomy are discussed with a separate discussion on the potential complications of using radiofrequency energy for meniscal ablation, including articular cartilage damage, osteonecrosis, and damage caused by irrigant. CONCLUSION: Electrosurgery has been shown to be an effective tool in arthroscopic meniscectomy. Further research and refinement is warranted because it may show superiority to other methods in certain situations.

Arthroscopy↗

Pregnancy after liver transplantation.

The first known posttransplantation pregnancy was in 1958 in a renal transplant recipient who had received a kidney from her identical twin sister. The first known posttransplantation pregnancy in a liver transplant recipient was in 1978. Information available from female kidney transplant recipients helped in the decision making involved in the management of this case, as well as those that followed. Over the last 20 years, issues specific to liver transplantation and pregnancy have been identified. Similar to the kidney transplant recipient population, when prepregnancy recipient graft function is stable and adequate, pregnancy appears to be well tolerated. Also similar to kidney transplant recipients, there has been no evidence of a specific malformation pattern among the children, and although prematurity and low birth weight occur, overall newborn outcomes have been favorable. Pregnancy in the setting of recurrent liver disease, such as recurrent hepatitis C, poses a potential problem among liver transplant recipients, as well as the possible adverse effects of immunosuppression on maternal kidney function. Also of significance, peripartum graft deterioration has more severe consequences in this transplant recipient population. Therefore, pregnancy must be considered carefully in this transplant recipient group. Since 1991, the National Transplantation Pregnancy Registry (NTPR) has studied the safety of pregnancy outcomes in solid-organ transplant recipients. The purpose of this review is to catalog studies in the literature, as well as to present current data from the registry with management guidelines.

Female↗

Three-dimensional ultrasonography in early pregnancy.

Three-dimensional ultrasound was introduced into clinical use over a decade ago. Early attempts on three-dimensional ultrasound were primitive and limited to experimental use only. Recently, it was used as a powerful tool to display three-dimensional fetal appearance and assess normal and pathological conditions. Because of the improvement in image resolution and reconstruction speed, three-dimensional ultrasound is now more and more popular in many centers, and used commonly as an adjunct in prenatal diagnosis. Indeed, a skillful sonographer may usually make the correct diagnosis of fetal anomaly with simply two-dimensional ultrasound. Nonetheless, the situation is gradually changing, because some anomalies have been diagnosed only based on the findings of three-dimensional ultrasound. In spite of that, the application of three-dimensional ultrasound is still less discussed in early pregnancy. In this article, the authors clarify the capability of three-dimensional ultrasound in different anatomic areas, and catalog its clinical merits in the early pregnancy to date. We also present the safety guidelines for its use in the first trimester.

Embryonic and Fetal Development↗

Assessment of language structure: from syntax to event-based analysis.

The last 70 years have seen much research and clinical attention devoted to the descriptions and assessment of children's developing language structures. From these efforts have evolved a number of ways to describe the kinds of structures children evidence at different stages of development, and thus the means to make comparisons across children or across time periods. This article reviews major trends that have emerged in syntactic descriptions and approaches to language habilitation, culminating in contemporary approaches that view syntactic structure in real life events rather than in artificial tasks or with arbitrary checklists. These new approaches allow us to determine the communication goals that are not being met rather than cataloging the syntactic structures that have not yet been mastered and provide us with the contexts for helping the children develop structures that will lead to clearer expression of meanings and fewer breakdowns in communication.

Child↗

Molecular analysis of genes on Xp controlling Turner syndrome and premature ovarian failure (POF).

Monosomy X has been known to be the chromosomal basis of Turner syndrome (TS) for more than four decades. A large body of cytogenetic data indicates that most TS features are due to reduced dosage of genes on the short arm of the X chromosome (Xp). Phenotype mapping studies using molecular cytogenetic and genetic techniques are beginning to localize the Xp genes that are important for various TS features, and a comprehensive catalog of candidate genes is becoming available through the Human Genome Project and related research. It is now possible to assess the contributions of individual genes to the TS phenotype by mutational analysis of karyotypically normal persons with specific TS features. This strategy has succeeded in identifying a gene involved in short stature and is being applied to premature ovarian failure and other TS phenotypes.

Body Height↗

[Shoulder dystocia].

In the increasing number of plaintiffs against mal-practice shoulder dystocia and plexus paralysis take a special role. Legal decisions are leaning too one-sided on statistical experience of childrens weight. But form and diameter of the pelvis should also be considered and better registered in medical record. A catalog of risks is given to prevent obstetrical cause of shoulder dystocia and plexus paralysis. The paper refers to the social and psychological change in which the child-parents must have more right to decide between cesarean section or vaginal delivery to prevent injury of the baby.

Attitude of Health Personnel↗

[Validity of risk calculations in offenders -- criticism of a basic methodological assumption, and future perspectives].

Catalogs of multidimensional criteria have become established in continental Europe. In contrast, due to the limited reliability of clinical prognoses, procedures such as "Actuarial Risk Assessments" have been established in Anglo-Saxon countries as a means of assessing prognoses. The basic assumptions of "Actuarial Risk Assessments" (principally a statistical methodology) show a remarkable discrepancy compared with the fundamental ideas of the European experts (clinical assessment procedures), although this has not to date been discussed in depth in the literature. Apart from this discrepancy, however, certain basic assumptions are internationally regarded as generally accepted methodological cornerstones of prognostic theory. Thus, such a generally unchallenged "essential" methodological principle as the "Two by Two Table" has been mentioned for many years in a dichotomous sense in numerous textbooks or basic lecture on prognostic theory. Far-reaching conclusions concerning the validity of prognostic statements are made on the basis of this methodological axiom. Thus, the putative error rate for prognostic assessments is derived from so-called "false positive prognoses", and, among other things, the unjustified committal of offenders wrongly judged to be dangerous is criticised. Surprisingly, there is no discussion in the considerable literature currently available, that it is methodologically highly questionable to use the Two by Two Table that is adequate for dichotomous decision-making, without using further differentiation with respect to probability factors, as is undoubtedly the case for risk calculations. The article discusses the above mentioned problems and gives possible approaches for solutions.

Actuarial Analysis↗

Photodissociation dynamics of the NO dimer. I. Theoretical overview of the ultraviolet singlet excited states.

Molecular orbital theory and calculations are used to describe the ultraviolet singlet excited states of NO dimer. Qualitatively, we derive and catalog the dimer states by correlating them with monomer states, and provide illustrative complete active space self-consistent field calculations. Quantitatively, we provide computational estimates of vertical transition energies and absorption intensities with multireference configuration interaction and equations-of-motion coupled-cluster methods, and examine an important avoided crossing between a Rydberg and a valence state along the intermonomer and intramonomer stretching coordinates. The calculations are challenging, due to the high density of electronic states of various types (valence and Rydberg, excimer and charge transfer) in the 6-8 eV region, and the multiconfigurational nature of the ground state. We have identified a bright charge-transfer (charge-resonance) state as responsible for the broadband seen in UV absorption experiments. We also use our results to facilitate the interpretation of UV photodissociation experiments, including the time-resolved 6 eV photodissociation experiments to be presented in the next two papers of this series.

Journal Article↗