Iron(II)/reductant(DH2)-induced activation of dioxygen for the hydroxylation and ketonization of hydrocarbons; mimics for the cytochrome P-450 hydroxylase/reductase system

Several metal complexes [(FeII(DPAH)2 (DPAH2 = 2,6-dicarboxyl pyridine), FeII(PA)2 (PAH = picolinic acid), FeII(bpy)2(2+), FeII(OPPh3)4(2+), (Cl8TPP)FeIIIX (X = Cl, OH, SCH2Ph) [Cl8TPP = tetrakis (2,6-dichlorophenyl)porphyrin], (TPP) FeIIICl (TPP = tetraphenylporphyrin), and CuI(tpy)2+ (typ = 2,2'-6,2"-terpyridine)] in combination with one of several reductants [DH2; PhNHNHPh (mimic of dihydroflavin), PhNHNH2, ascorbic acid (H2asc), and PhCH2SH (model ligand for cysteine residue)] catalytically activate O2 (1 atm) for the hydroxylation of saturated hydrocarbons (e.g. c-C6H12-->c-C6H11OH). This chemistry closely parallels that of cytochrome P-450 proteins, and both appear to involve a Fenton-like reactive intermediate), [LxFeOOH(DH)]. With cyclohexane as the substrate the dominant product is its ketone (as well as significant amounts of its hydroperoxide). 1,4-Cyclohexadiene (with two double-allylic carbon centers) undergoes dehydrogenation to give benzene, but also yields substantial amounts of phenol via ketonization of an allylic carbon. The 1:1 FeII(bpy)2(2+)/(PhNHNH2 or H2asc), FeII(PA)2/H2asc, and (Cl8TPP)FeIIICl/PhNHNH2 combinations initiate the autoxidation of 1,4-cyclohexadiene with turnover numbers (moles of product per mole of reductant) from 71 to 26, respectively (alpha-tocophenol reduces the turnover numbers by 20-80%). With respect to aerobic biology, the present results indicate that dysfunctional transition metals (degradation products of metalloproteins) in combination with biological reductants activate O2 for reaction with organic substrates. The level of activation is similar to that for Fenton reagents and cytochrome P-450 hydroxylases. Hence, dysfunctional transition metals, reductants, and O2 are a hazardous combination within a biological matrix.     

Antithrombin III and trauma patients: factors that determine low levels

Using capillary videomicroscopy of the nail fold, the frequency of cold-induced vasospasm and capillary hemodynamic parameters were studied after application of cold in 50 patients with primary fibromyalgia, 50 patients with chronic low back pain, and 50 healthy controls. Cold-induced vasospasm was detected in 38% of the patients with fibromyalgia. In this group it was significantly more frequent than in the patients with chronic low back pain (20%, p < 0.05) and healthy subjects (8%, p < 0.001). In the fibromyalgia group, the magnitude of vasospasm as measured by the capillary blood flow deceleration after cold application correlated negatively with the pain intensity as measured by pain score (r = -0.3839, p < 0.01). No differences in clinical appearance were found between patients with and without cold-induced vasospasm in both the fibromyalgia and low back pain group.     

One world, one hope: the cost of providing antiretroviral therapy to all nations

OBJECTIVE: To estimate the potential direct cost of making triple combination antiretroviral therapy widely available to HIV-positive adults and children living in countries throughout the world. METHODS: For each country, antiretroviral costs were obtained by multiplying the annual cost of triple antiretroviral therapy by the estimated number of HIV-positive persons accessing therapy. Per capita antiretroviral costs were computed by dividing the antiretroviral costs by the country's total population. The potential economic burden was calculated by dividing per capita antiretroviral costs by the gross national product (GNP) per capita. All values are expressed in 1997 US dollars. RESULTS: The potential cost of making triple combination antiretroviral therapy available to HIV-positive individuals throughout the world was estimated to be over US$ 65.8 billion. By far the greatest financial burden was on sub-Saharan Africa. The highest per capita drug cost in this region would be incurred in the subregions of Southern Africa (US$ 149) followed by East Africa (US$ 116), Middle Africa (US$ 44), and West Africa (US$ 42). In the Americas, subregional data indicated the highest per capita drug cost would be in the Latin Caribbean (US$ 22), followed by the Caribbean (US$ 17), Andean Area (US$ 7), the Southern Cone (US$ 6), North America (US$ 6), and Central American Isthmus (US$ 5). In Asia and Europe the percentage of the GNP necessary to finance drug therapy was less than 1% in most countries examined. CONCLUSION: Our results demonstrate that the cost of making combination antiretroviral therapy available worldwide would be exceedingly high, especially in countries with limited financial resources.     

Interleukin (IL)-10, IL-12, and interferon-gamma production in primary and memory immune responses to varicella-zoster virus

Varicella immunization provided the opportunity to examine the kinetics of interleukin (IL)-10, IL-12 and interferon (IFN)-gamma production elicited during primary in vivo sensitization with proteins of varicella-zoster virus (VZV), a common human herpesvirus. VZV-specific IFN-gamma release and T cell proliferation were elicited by immunization and persisted through 15 months of follow-up. The induction of VZV-specific T cells and IgG antibodies was accompanied by transient increases in IL-10 and IL-12 production. T cell proliferation to VZV was significantly lower in adults at 15 months than in vaccinated children or naturally immune subjects and correlated with lower IFN-gamma responses in individual vaccinees. After primary immunity was induced, continued IL-12 production was not necessary to maintain the predominant Th1-type response elicited by VZV. Cytokine profiles observed during primary in vivo sensitization to VZV suggest that parallel increases in IFN-gamma and IL-10 may be important in the induction of immunity to some viral pathogens.     

The anatomy of a hypoxic operator in Saccharomyces cerevisiae

Aerobic repression of the hypoxic genes of Saccharomyces cerevisiae is mediated by the DNA-binding protein Rox1 and the Tup1/Ssn6 general repression complex. To determine the DNA sequence requirements for repression, we carried out a mutational analysis of the consensus Rox1-binding site and an analysis of the arrangement of the Rox1 sites into operators in the hypoxic ANB1 gene. We found that single base pair substitutions in the consensus sequence resulted in lower affinities for Rox1, and the decreased affinity of Rox1 for mutant sites correlated with the ability of these sites to repress expression of the hypoxic ANB1 gene. In addition, there was a general but not complete correlation between the strength of repression of a given hypoxic gene and the compliance of the Rox1 sites in that gene to the consensus sequence. An analysis of the ANB1 operators revealed that the two Rox1 sites within an operator acted synergistically in vivo, but that Rox1 did not bind cooperatively in vitro, suggesting the presence of a higher order repression complex in the cell. In addition, the spacing or helical phasing of the Rox1 sites was not important in repression. The differential repression by the two operators of the ANB1 gene was found to be due partly to the location of the operators and partly to the sequences between the two Rox1-binding sites in each. Finally, while Rox1 repression requires the Tup1/Ssn6 general repression complex and this complex has been proposed to require the aminoterminal regions of histones H3 and H4 for full repression of a number of genes, we found that these regions were dispensable for ANB1 repression and the repression of two other hypoxic genes.     

The possibility of preserving the causative agent of plague in soil in resting (nonculturable) form

The purpose of this study was to find criteria characteristic for patients in need of care and social services. The criteria should serve as a guideline for patients and staff to facilitate care planning before discharge. The sample consisted of 49 patients, born before 1925, in need of emergency inpatient treatment, admitted to medical- or orthopaedic wards. Data of the patient's self care needs were collected by interviews, assessment of self care status and need of treatment. The patients could be divided into three groups depending on type of discharge. Group A (n = 27) discharged home, group B (n = 7) discharged to geriatric clinic and group C (n = 15) discharged and in need of further care and social services. Criteria indicating the patients further assistance from the community were in group C (medical- and orthopaedic wards) deficit in daily living activities and locomotion. Group B had an increased need of support from the physiotherapist and the occupational therapist, in locomotion as well as daily living activities The physician's assessment showed that the criteria behind the decision "no further medical treatment appropriate" and "ready for discharge" were not related to medical impairment but to lack of self care, need of care, rehabilitation and social services.     

Age-Dependent Effects of Jasmonic Acid Treatment and Wind Exposure on Foliar Oxidase Activity and Insect Resistance in Tomato

Jasmonic acid (JA) treatment of tomato plants induces several defense-related oxidative enzymes and increases pest resistance in a manner thought to simulate natural insect wounding. In a full-factorial greenhouse experiment, we examined the independent and interactive effects of plant age and exposure to wind-induced mechanical stress (MS), on the ability of JA to induce defense in tomato. In general, treatment of 4-, 6-, and 8-week-old tomato plants with 1 mM JA resulted in the induction of peroxidase and polyphenol oxidase activity and reduced the relative growth rate of first-instar Manduca sexta larvae fed treated leaves, in accordance with other studies. Peroxidase activity increased with plant age and was induced by JA most strongly in older plants. In contrast, polyphenol oxidase activity did not change with plant age and was induced by JA most strongly in young plants. While relative growth rates of M. sexta were lower on older plants overall, JA reduced growth rates most strongly in young plants, in which JA treatment enhanced polyphenol oxidase activity by more than 70%. MS enhanced the activity of peroxidase, but substantially reduced the activity of polyphenol oxidase; the latter most intensely on older plants. M. sexta tended to grow more slowly on MS-treated plants, although this effect was not significant. Thus, reduced polyphenol oxidase activity in MS-treated plants did not lead to an increase in growth rate of M. sexta, possibly because peroxidase activity was still elevated in MS-treated plants. Significant interactions between JA and MS and three-way interactions were not detected for any variable, although the inductive effects of both JA and MS interacted in complex ways with plant age. Our results indicate that resistance traits in tomato are differentially affected by JA and wind exposure and differ in their relative contribution to defense as plants age.     

DNA vaccination against Theiler''s murine encephalomyelitis virus leads to alterations in demyelinating disease

Using glycerinated spasmoneme of giant Zoothamnium sp., the physical properties of spasmoneme before and after Ca2+-induced contraction (pCa 4.5) were investigated. The volume change of spasmoneme contraction was measured under zero tension. The length and diameter decreased by about 50% of their initial value as a result of contraction, which means that contraction is nearly isotropic. Thus the volume of spasmoneme decreased drastically by 86% of its original value. The swollen ratio of extended and contracted spasmoneme were 0.07 and 0.37, respectively. Tension-extension relationships of extended and contracted spasmonemes were measured. By applying the theory of rubber elasticity, the number of segments of a chain in originally extended spasmoneme was only 3.3, i.e., the chain was almost a straight one. On the other hand, the number of segments of a chain in contracted spasmoneme was more than 100, i.e., the chain was essentially a random one. Furthermore, the total number of chains in single spasmoneme was the same in extended and contracted spasmoneme. This means that the interchain cross-links of chains were not influenced by addition or removal of Ca2+. Moreover, the molecular weight of a chain is estimated to be at most about 50 kd. By considering all these results, it is concluded that the contractile mechanism of spasmoneme originates in the intramolecular folding and unfolding induced by Ca2+ binding and detaching.