Formation Constants and Conformational Analysis of Carbamates in Aqueous Solutions of 2-Methylpiperidine and CO2 from 283 to 313 K by NMR Spectroscopy.

Quantitative 13C nuclear magnetic resonance (NMR) spectroscopy was used to investigate the speciation in (2-methylpiperidine + H2O + CO2) systems at 283.2-313.2 K. The carbamate of 2-methylpiperidine(2-methylpiperidine- N-carboxylate) was shown for the first time to be a stable species in aqueous solutions. The spectroscopic results were used to obtain temperature-dependant formation constants for the carbamate using a simplified model for the activity coefficients from which the standard molar enthalpy of reaction was estimated. The results were incorporated into a self-consistent chemical equilibrium model, which includes vapor-liquid equilibria and all aqueous species, including the formation of carbamate. The predominant conformation of the sterically hindered carbamate, which was determined using two-dimensional exchange spectroscopy NMR, has the methyl group in the axial orientation and is in agreement with the density functional theory quantum chemical calculations.

Lumbar magnetic resonance imaging findings in patients with and without Waddell Signs.

BACKGROUND CONTEXT: Waddell Signs (WS), introduced as a method to establish patients with substantial psychosocial components to their low back pain, carry a negative association despite no literature evaluating whether physical disease is associated with them. PURPOSE: To compare lumbar magnetic resonance imaging (MRI) findings between the patients with and without WS. STUDY DESIGN: Retrospective cohort study based on prospectively collected data. PATIENT SAMPLE: Thirty patients aged 35 to 55 years with an Oswestry Disability Index (ODI) score >50 randomly selected such that there was an even distribution of patients based on the number of WS. OUTCOME MEASURES: ODI and Short Form-12 scores, number of WS, presence and severity of spinal pathology. METHODS: MRIs were reviewed by three spine specialists blinded to clinical exam findings, number of WS, and patient identity. Type and severity of pathology and presence of surgical and non-surgical lesions were assessed, and findings were rank ordered based on the overall impression of the pathology. There was no external funding or potential conflicts of interest for this study. RESULTS: There were significantly more individual pathologic findings in patients without WS (p=.02). However, there was no difference in the severity of pathology based on WS (p=.46). Furthermore, the rank ordering based on overall impression of severity showed no difference between the patients with and without WS (p=.20). Although 100% of the patients without WS showed pathologic findings on MRI, 70% of WS patients also had significant pathology on MRI. The prevalence of spondylolisthesis, stenosis, and disc herniation was similar (p=.41, p=.22, and p=.43, respectively). The prevalence and mean number of lesion amenable to surgery did not differ based on presence of WS (p=.21 and p=.18, respectively). CONCLUSIONS: Patients with WS present a difficult diagnostic challenge for the physician as their organic symptoms are often coexistent with emotional fear avoidance behavior. Although there is more overall pathology in patients without WS, a significant number of these patients appear to have comparable spinal pathology with equivalent severity, which may be contributing to patients' symptoms and disability. Presence of these non-organic symptoms often makes us doubt these patients. However, as part of effective treatment, physicians should better understand both the physical and psychological components of patient disability.

Early effects of climate change: do they include changes in vector-borne disease?

The world's climate appears now to be changing at an unprecedented rate. Shifts in the distribution and behaviour of insect and bird species indicate that biological systems are already responding to this change. It is well established that climate is an important determinant of the spatial and temporal distribution of vectors and pathogens. In theory, a change in climate would be expected to cause changes in the geographical range, seasonality (intra-annual variability), and in the incidence rate (with or without changes in geographical or seasonal patterns). The detection and then attribution of such changes to climate change is an emerging task for scientists. We discuss the evidence required to attribute changes in disease and vectors to the early effects of anthropogenic climate change. The literature to date indicates that there is a lack of strong evidence of the impact of climate change on vector-borne diseases (i.e. malaria, dengue, leishmaniasis, tick-borne diseases). New approaches to monitoring, such as frequent and long-term sampling along transects to monitor the full latitudinal and altitudinal range of specific vector species, are necessary in order to provide convincing direct evidence of climate change effects. There is a need to reassess the appropriate levels of evidence, including dealing with the uncertainties attached to detecting the health impacts of global change.

Pyrazinamide inhibits the eukaryotic-like fatty acid synthetase I (FASI) of Mycobacterium tuberculosis.

Tuberculosis treatment is shortened to six months by the indispensable addition of pyrazinamide (PZA) to the drug regimen that includes isoniazid and rifampin. PZA is a pro-drug of pyrazinoic acid (POA) (ref. 3), whose target of action has never been identified. Although PZA is active only against Mycobacterium tuberculosis, the PZA analog 5-chloro-pyrazinamide (5-Cl-PZA) displays a broader range of anti-mycobacterial activity. We have found that the eukaryotic-like fas1 gene (encoding fatty acid synthetase I, FASI) from M. avium, M. bovis BCG or M. tuberculosis confers resistance to 5-Cl-PZA when present on multi-copy vectors in M. smegmatis. 5-Cl-PZA and PZA markedly inhibited the activity of M. tuberculosis FASI, the biosynthesis of C16 to C24/C26 fatty acids from acetyl-CoA (ref. 6). Importantly, PZA inhibited FASI in M. tuberculosis in correlation with PZA susceptibility. These results indicate that FASI is a primary target of action for PZA in M. tuberculosis. Further characterization of FASI as a drug target for PZA may allow the development of new drugs to shorten the therapy against M. tuberculosis and may provide more options for treatment against M. bovis, M. avium and drug resistant M. tuberculosis.

A novel mycolic acid cyclopropane synthetase is required for cording, persistence, and virulence of Mycobacterium tuberculosis.

Colonial morphology of pathogenic bacteria is often associated with virulence. For M. tuberculosis, the causative agent of tuberculosis (TB), virulence is correlated with the formation of serpentine cords, a morphology that was first noted by Koch. We identified a mycobacterial gene, pcaA, that we show is required for cording and mycolic acid cyclopropane ring synthesis in the cell wall of both BCG and M. tuberculosis. Furthermore, we show that mutants of pcaA fail to persist within and kill infected mice despite normal initial replication. These results indicate that a novel member of a family of cyclopropane synthetases is necessary for lethal chronic persistent M. tuberculosis infection and define a role for cyclopropanated lipids in bacterial pathogenesis.

Complex lipid determines tissue-specific replication of Mycobacterium tuberculosis in mice.

Tuberculosis is the leading cause of death in the world resulting from a single bacterial infection. Despite its enormous burden on world health, little is known about the molecular mechanisms of pathogenesis of Mycobacterium tuberculosis. Bacterial multiplication and concomitant tissue damage within an infected host, including experimentally infected mice, occurs primarily in the lungs-the favoured niche of M. tuberculosis. Although it has been proposed that the distinctive cell wall of M. tuberculosis is important for virulence, rigorous genetic proof has been lacking. Here, using signature-tagged mutagenesis, we isolated three attenuated M. tuberculosis mutants that cannot synthesize or transport a complex, cell wall-associated lipid called phthiocerol dimycocerosate (PDIM) which is found only in pathogenic mycobacteria. Two mutants have transposon insertions affecting genes implicated in PDIM synthesis; the third has a disruption in a gene encoding a large transmembrane protein required for proper subcellular localization of PDIM. Synthesis and transport of this complex lipid is only required for growth in the lung; all three mutants are unaffected for growth in the liver and spleen. This clearly shows that a lipid is required for M. tuberculosis virulence.

Global assessment of El Niño's disaster burden.

BACKGROUND: Natural disasters have profound effects on health and require medical intervention as part of relief operations. The world's populations are becoming increasingly vulnerable to extreme weather events, which are responsible for most natural disasters. The El Niño Southern Oscillation (ENSO) is the most prominent global climate system associated with year-to-year weather variability and extreme events. We have estimated the burden on human health of natural disasters associated with ENSO. METHODS: We used time-series regression analysis of the relation between El Niño years and the annual rates of persons affected by natural disasters per 1000 population during 1964-93, globally and also by region and disaster type. Correlations between sea-surface temperature (SST) anomalies (index of ENSO) and the rates of persons affected by natural disasters per 1000 population were determined globally, by region and by disaster type. FINDINGS: The rate of persons affected by natural disasters worldwide is strongly associated with ENSO; rates are greater during the first El Niño year (p = 0.05) and the following year (p = 0.01) than in the pre-Niño year. The correlation between rates of persons affected by natural disasters and SST anomalies in the Eastern Pacific (a key ENSO indicator) is highest in the last quarter of the previous year (r = 0.53, p < 0.01). These associations are strongest in South Asia, the region where more than 50% of all disaster victims live. Worldwide, rates of persons affected by drought/famine (half of all disaster victims) and by volcanic eruptions show significant associations with the ENSO cycle, being highest in the post-Niño year and El Niño year, respectively, and being significantly associated with SST anomalies. INTERPRETATION: The strong relation between ENSO and populations affected by natural disasters can be described as a "natural disaster cycle". Determining the phase in this cycle, using SST from the Eastern Equatorial Pacific, could benefit disaster preparedness on a global scale, for South Asia in particular, and for all populations affected by drought/famine and volcanic disasters.

The unfolded protein response coordinates the production of endoplasmic reticulum protein and endoplasmic reticulum membrane.

The endoplasmic reticulum (ER) is a multifunctional organelle responsible for production of both lumenal and membrane components of secretory pathway compartments. Secretory proteins are folded, processed, and sorted in the ER lumen and lipid synthesis occurs on the ER membrane itself. In the yeast Saccharomyces cerevisiae, synthesis of ER components is highly regulated: the ER-resident proteins by the unfolded protein response and membrane lipid synthesis by the inositol response. We demonstrate that these two responses are intimately linked, forming different branches of the same pathway. Furthermore, we present evidence indicating that this coordinate regulation plays a role in ER biogenesis.

A novel mechanism for regulating activity of a transcription factor that controls the unfolded protein response.

Cells respond to an accumulation of unfolded proteins in the endoplasmic reticulum (ER) by increasing transcription of genes encoding ER-resident proteins. The information is transmitted from the ER lumen to the nucleus by an intracellular signaling pathway, the unfolded protein response (UPR). We have identified a basic-leucine zipper transcription factor, Hac1p, that is required for the UPR and binds to the UPR element in the promoter of UPR-regulated genes. Surprisingly, Hac1p is found in UPR-activated cells only, and its level is controlled by regulated splicing of its mRNA. Splicing replaces the C-terminal tail of Hac1p with a different peptide that renders Hac1p more resistant to an otherwise extremely rapid ubiquitin-dependent degradation. We propose that the complex regulation of Hac1p expression serves to provide multiple levels at which the UPR can be controlled.

tRNA ligase is required for regulated mRNA splicing in the unfolded protein response.

The accumulation of unfolded proteins in the endoplasmic reticulum (ER) triggers an intracellular signaling pathway, the unfolded protein response (UPR), that leads to increased transcription of genes encoding ER-resident proteins. Transcriptional activation is mediated by a dedicated transcription factor, Hac1p, whose activity is controlled by regulated splicing of its mRNA. We have identified a mutation in tRNA ligase that disrupts the UPR in the yeast Saccharomyces cerevisiae. In this mutant, splicing of HAC1 mRNA, but not tRNA, is blocked. In contrast, HAC1 mRNA splicing is not impaired in cells that are blocked in spliceosome-mediated mRNA splicing. Furthermore, the splice junctions of HAC1 mRNA do not conform to the consensus sequences of other yeast pre-mRNAs. Our results suggest that the regulated splicing of HAC1 mRNA occurs by a novel pathway, involving tRNA ligase and bypassing the spliceosome.

Environmental fate of rice pesticides in California.

Each of the pesticides reviewed is reported to dissipate from field water after application. Carbofuran is hydrolyzed rapidly under the alkaline conditions usually found in the rice field environment, and its hydrolysis products are also degraded rapidly. The longest half-life reported (18-26 d) was in water that overlaid soil treated with Furadan granular formulation. Generally, carbofuran dissipation ranged from 36 hr to 3 d. Under field application, bensulfuron methyl showed a half-life of 1-3 d, but others have recovered all of the "dissipated" herbicide in the soil compartment. MCPA applied to rice fields is reportedly degraded by the joint action of sunlight and microbial action with a half-life of 3-5 d. Methyl parathion showed a maximum half-life of 9-17 d in a model aquatic ecosystem, but other reports found more rapid dissipation. The half-life of molinate has been observed by numerous researchers to be less than 5 d, with volatility the major route of loss. A half-life as short as 5-7 d has been reported for thiobencarb applied to rice fields, but others report much longer periods; volatility again is expected to be a significant route of loss. Microbial degradation takes place with each of the subject pesticides. Numerous authors have reported enhanced degradation of carbofuran under conditions of repeated application, and this probably holds true for the others. A specialized segment of the microbial population (Pseudomonas spp.) is purported to carry out most of the degradation but is inefficient at degrading the hydrolysis product, carbofuran phenol. Biodegradation of bensulfuron methyl has been observed with actinomycetes, fungi, and bacteria, and takes place primarily by oxidation and hydrolysis. Methyl parathion is biodegraded primarily by nitro reduction to aminomethyl parathion. A fungus, an actinomycete, and a bacterium were shown to biodegrade molinate, primarily by oxidation at the sulfur atom and the azepine ring. Thiobencarb is biodegraded in anaerobic sediments at a slow rate; the dechlorinated thiobencarb was shown to cause dwarfing of rice in some fields. Otherwise, its aerobic biodegradation is rapid and follows the same routes as with molinate. Carbofuran is a systemic insecticide that is rapidly absorbed and translocated to aerial parts of the plant. Carbofuran is metabolized in rice plants to the corresponding phenol, which is irreversibly bound into the plant, as well as to 3-hydroxycarbofuran and other minor components; it is depurated through leaf exudate, from which it volatilizes. Rice plants were observed to take up more bensulfuron methyl through shoots than roots and to metabolize it to the 4-hydroxy analog. The half-life of methyl parathion in Hydrilla verticulla, an aquatic macrophyte, was 1 wk, but little has been reported on methyl parathion in plants. Barnyardgrass was found to absorb greater amounts of molinate than did rice, and it produced larger proportions of basic metabolites, which may form the basis for its selective toxicity. Thiobencarb has been shown to be rapidly absorbed, translocated, and metabolized in rice plants, barnyardgrass, and the broadleaved wild amaranth, smartweed, and lambsquarters. Translocation was more rapid and extensive in barnyardgrass than in rice, and most of the 14C radiolabel was recovered as metabolites. Its terminal metabolite, chlorobenzoic acid, was taken up into lignin-like plant constituents. It is apparent that information on plant uptake and biodegradation is limited--none exists for woody species--but the fact that some species appear resistant to the herbicides suggests that biodegradative ability is general. (ABSTRACT TRUNCATED)

Orthopaedic complications of childhood neurofibromatosis.

Neurofibromatosis is a genetically transmitted, multisystemic disorder characterized by abnormalities of the skin, nervous tissue, and bone. Many of the serious problems associated with this disease are orthopaedic in nature. The treatment and plan of care for a child with neurofibromatosis must be comprehensive and consistent. Nursing care should address the developmental, physical, and emotional needs of the child, as well as the psychologic concerns and educational needs of the child and parents.

Orthopaedic complications of childhood neurofibromatosis.

Neurofibromatosis is a genetically transmitted, multisystemic disorder characterized by abnormalities of the skin, nervous tissue, and bone. Many of the serious problems associated with this disease are orthopaedic in nature. The treatment and plan of care for a child with neurofibromatosis must be comprehensive and consistent. Nursing care should address the developmental, physical, and emotional needs of the child, as well as the psychologic concerns and educational needs of the child and parents.

Laparoscopic versus open appendicectomy for suspected appendicitis: a prospective study.

Despite recent advances in minimally invasive surgery, laparoscopic appendicectomy has been questioned as a feasible method of treating patients with suspected appendicitis because open appendicectomy carries few risks and complications. Between February 1992 and January 1993 a non-randomized prospective study comparing open and laparoscopic appendicectomy was designed to assess differences in postoperative morbidity, pain, inpatient hospital stay and a return to normal lifestyle. One hundred and sixty-seven patients with suspected clinical appendicitis were assigned to open (n = 74) or laparoscopic (n = 93) appendicectomy. Six patients were excluded due to the presence of other significant pathology such that the appendix was not removed. Eighty-seven patients underwent attempted laparoscopic appendicectomy, of which 81 were completed. The diagnosis of appendicitis was confirmed histologically in 63.5% of patients undergoing open appendicectomy and 63% of patients undergoing laparoscopic appendicectomy. There were no significant differences in anaesthetic times, postoperative morbidity and analgesic requirements. There was a significant reduction in both postoperative inpatient stay (P < 0.0001) and time taken to return to work or normal activities (P < 0.0001) for the laparoscopic group. The use of laparoscopy for patients with suspected appendicitis aids definitive diagnosis and should reduce the negative appendicectomy rate to an acceptable figure. The results suggest that laparoscopic appendicectomy is safe and offers advantages over open appendicectomy in the management of patients with suspected appendicitis.

The unfolded-protein-response pathway in yeast.

The accumulation of unfolded proteins in the endoplasmic reticulum (ER) triggers the increased production of several ER-resident proteins. This signalling pathway exists in organisms as divergent as mammals and yeast, and is the only known example of an intracellular signalling system that links the ER and the nucleus. Recently, a transmembrane kinase similar in structure to growth-factor receptor kinases has been identified as a key component of the unfolded-protein-response pathway in yeast.

Transcriptional induction of genes encoding endoplasmic reticulum resident proteins requires a transmembrane protein kinase.

The transcription of genes encoding soluble proteins that reside in the endoplasmic reticulum (ER) is induced when unfolded proteins accumulate in the ER. Thus, an intracellular signal transduction pathway must exist that mediates communication between the ER lumen and the nucleus. We have identified a gene in S. cerevisiae, IRE1, that is required for this pathway: ire1- mutants cannot activate transcription of KAR2 and PDI1, which encode the ER resident proteins BiP and protein disulfide isomerase. Moreover, IRE1 is essential for cell viability under stress conditions that cause unfolded proteins to accumulate in the ER. IRE1 encodes a transmembrane serine/threonine kinase that we propose transmits the unfolded protein signal across the ER or inner nuclear membrane. IRE1 is also required for inositol prototrophy, suggesting that the induction of ER resident proteins is coupled to the biogenesis of new ER membrane.

A promoter element that exerts positive and negative control of the interleukin 2-responsive J-chain gene.

In a primary immune response a signal from interleukin 2 (IL-2) induces B lymphocytes to express the gene for the IgM joining component, the J chain. The signaling mechanism was pursued in this study by examining the J-chain gene 5' flanking region for regulatory sequences and interacting nuclear factors. The analyses identified a major control region located between -75 and -45 that encodes two adjacent elements: a T-rich sequence (JA) containing a single positive regulatory motif and an A+G-rich sequence (JB) containing overlapping positive and negative regulatory motifs. Dissection of the two elements indicated that the bifunctional JB sequence is the likely target of the IL-2 signal. The evidence was based on findings that (i) JB activity correlated with J-chain gene transcription--i.e., JB acts as a repressor in J-chain-silent B cells and as an activator in J-chain-expressing cells, and (ii) JB activator function is mediated by a B-cell-specific nuclear protein, NF-JB, that exhibits an IL-2-responsive binding pattern.

Long-term outcome of a self-help very-low-calorie-diet weight-loss program.

This paper presents 90-wk data on five seriously overweight subjects originally brought together for 1-2 wk in simple residential accommodation to share the experience of beginning a program of very-low-calorie dieting. All subjects have remained well below their starting weight; four of the five subjects have continued to lose weight (weight losses 33.7-66.8 kg), now weighing less than at the end of the first 26 wk previously reported (1). It is proposed that this self-help group model, incorporating a very-low-calorie diet, is valuable for the long-term management of obesity.