Alterations of Choroidal Blood Flow Regulation in Young Healthy Subjects with Complement Factor H Polymorphism

A common polymorphism in the complement factor H gene (rs1061170, Y402H) is associated with a high risk of age-related macular degeneration (AMD). In the present study we hypothesized that healthy young subjects homozygous for the high-risk haplotype (CC) show abnormal choroidal blood flow (ChBF) regulation decades before potentially developing the disease. A total of 100 healthy young subjects were included in the present study, of which 4 subjects were excluded due to problems with genotyping or blood flow measurements. ChBF was measured continuously using laser Doppler flowmetry while the subjects performed isometric exercise (squatting) for 6 minutes. The increase in ChBF was less pronounced than the response in ocular perfusion pressure (OPP), indicating for some degree of choroidal blood flow regulation. Eighteen subjects were homozygous for C, 47 subjects were homozygous for T and 31 subjects were heterozygous (CT). The increase in OPP during isometric exercise was not different between groups. By contrast the increase in ChBF was more pronounced in subjects homozygous for the high risk C allele (p = 0.041). This was also evident from the pressure/flow relationship, where the increase in ChBF in homozygous C carriers started at lower OPPs as compared to the other groups. Our data indicate that the regulation of ChBF is abnormal in rs1061170 CC carriers. So far this polymorphism has been linked to age related macular degeneration (AMD) mainly via inflammatory pathways associated with the complement system dysfunction. Our results indicate that it could also be related to vascular factors that have been implicated in AMD pathogenesis.     

Involvement of cholesterol, calcium and progesterone in the induction of capacitation and acrosome reaction of mammalian spermatozoa

The aim of this paper was to review the effects of some important substances involved in the induction of sperm plasma membrane changes referred to as acrosome reaction, namely cholesterol (C), calcium (Ca(2+)) and progesterone (P(4)). For this purpose, mechanisms of capacitation and acrosome reaction (AR) as well as the processes, C, Ca(2+) and P(4) are involved in, are described. Subsequently, to get a better insight into possible beneficial and detrimental effects on sperm function, the occurrence of these molecules in semen extenders is discussed.     

High dissociation rate constant of ferrous-dioxy complex linked to the catalase-like activity in lactoperoxidase

Heme reduction of ferric lactoperoxidase (LPO) into its ferrous form initially leads to the accumulation of the unstable form of LPO-Fe(II), which spontaneously converts to a more stable species, the two of which can be identified by Soret peaks at 440 and 434 nm, respectively. Our data demonstrate that both LPO-Fe(II) species are capable of binding O(2) at a similar rate to generate the ferrous-dioxy complex. Its formation with respect to O(2) was first order and monophasic and with rate constants of k(on) = 3.8 x 10(4) m(-1) s(-1) and k(off) = 11.2 s(-1). The dissociation rate constant for the formation of LPO-Fe(II)-O(2) is relatively high, in contrast to hemoprotein model compounds. This high dissociation rate can be attributed to a combination of effects that include the positive trans effect of the proximal ligand, the heme pocket environment, and the geometry of the Fe-O(2) linkage. Our results have also shown that the decay of the LPO-Fe(II)-O(2) complex occurs by two sequential O(2)-independent steps. The first step involves formation of a short-lived intermediate that can be characterized by its Soret absorption peak at 416 nm and may be attributed to the weakening of the Fe(II)-O(2) linkage with a rate constant of 0.5 s(-1). The second step is spontaneous conversion of this intermediate to generate the native enzyme and presumably superoxide as end products with a rate constant of 0.03 s(-1). A comprehensive kinetic model that links LPO-Fe(II)-O(2) complex formation to the LPO catalase-like activity, combined with the classic catalytic cycle, is presented here.     

The potential role of nitric oxide in substrate switching in eosinophil peroxidase

Eosinophil recruitment and enhanced nitric oxide (NO) production are characteristic features of asthma and other airway diseases. Eosinophil peroxidase (EPO), a highly cationic hemoprotein secreted by activation of eosinophils, is believed to play a central role in host defense against invading pathogens. The enzyme uses hydrogen peroxide (H2O2) and bromide (Br-), a preferred cosubstrate of EPO, to generate the cytotoxic oxidant hypobromous acid. The aim of this work was to determine whether NO can compete with plasma levels of Br- and steer the enzyme reaction from a 2e- oxidation to a 1e- oxidation pathway. Rapid kinetic measurements were utilized to measure the rate of EPO compounds I and II formation, duration, and decay at 412 and 432 nm, respectively, at 10 degrees C. An EPO-Fe(III) solution supplemented with increasing Br- concentrations was rapidly mixed with fixed amounts of H2O2 in the absence and in the presence of increasing NO concentrations. In the absence of NO, EPO-Fe(III) primarily converted to compound I and, upon H2O2 exhaustion, it decayed rapidly to the ferric form. NO caused a significant increase in the accumulation of EPO compound II, along with a proportional increase in its rate of formation and duration as determined by the time elapsed during catalysis. The time courses for these events have been incorporated into a comprehensive kinetic model. Computer simulations carried out supported the involvement of a conformational intermediate in the EPO compound II complex decay. Collectively, our results demonstrated that NO displays the potential capacity to promote substrate switching by modulating substrate selectivity of EPO.     

Climatic requirements of the eastern paralysis tick,Ixodes holocyclus,with a consideration of its possible geographic range up to 2090

The eastern paralysis tick, Ixodes holocyclus, is an ectoparasite of medical and veterinary importance in Australia. The feeding of I. holocyclus is associated with an ascending flaccid paralysis which kills many dogs and cats each year, with the development of mammalian meat allergy in some humans, and with the transmission of Rickettsia australis (Australian scrub typhus) to humans. Although I. holocyclus has been well studied, it is still not known exactly why this tick cannot establish outside of its present geographic distribution. Here, we aim to account for the presence as well as the absence of I. holocyclus in regions of Australia. We modelled the climatic requirements of I. holocyclus with two methods, CLIMEX, and a new envelope-model approach which we name the ‘climatic-range method’. These methods allowed us to account for 93% and 96% of the geographic distribution of I. holocyclus, respectively. Our analyses indicated that the geographic range of I. holocyclus may not only shift south towards Melbourne, but may also expand in the future, depending on which climate-change scenario comes to pass.     

Melatonin is a potent inhibitor for myeloperoxidase

Myeloperoxidase (MPO) catalyzes the formation of potent oxidants that have been implicated in the pathogenesis of various diseases including atherosclerosis, asthma, arthritis, and cancer. Melatonin plays an important part in the regulation of various body functions including circadian sleep rhythms, blood pressure, oncogenesis, retinal function, seasonal reproduction, and immunity. Here, we demonstrate that melatonin serves as a potent inhibitor of MPO under physiological-like conditions. In the presence of chloride (Cl-), melatonin inactivated MPO at two points in the classic peroxidase cycle through binding to MPO to form an inactive complex, melatonin-MPO-Cl, and accelerating MPO compound II formation, an inactive form of MPO. Inactivation of MPO was mirrored by the direct conversion of MPO-Fe(III) to MPO compound II without any sign of compound I accumulation. This behavior indicates that melatonin binding modulates the formation of MPO intermediates and their decay rates. The Cl- presence enhanced the affinity of MPO toward melatonin, which switches the enzyme activity from peroxidation to catalase-like activity. In the absence of Cl-, melatonin served as a 1e- substrate for MPO compound I, but at higher concentration it limited the reaction by its dissociation from the corresponding complex. Importantly, melatonin-dependent inhibition of MPO occurred with a wide range of concentrations that span various physiological and supplemental ranges. Thus, the interplay between MPO and melatonin may have a broader implication in the function of several biological systems. This dual regulation by melatonin is unique and represents a new means through which melatonin can control MPO and its downstream inflammatory pathways.     

The development of myeloperoxidase inhibitors

Myeloperoxidase (MPO), an abundant hemoprotein present in neutrophils and monocytes, plays a significant role in immune surveillance and host defense mechanisms. However, increased MPO activity has been linked to a number of pathologies with compelling evidence in initiation and progression of inflammatory events. As a result, search for active compounds that can efficiently inhibit MPO activity and subsequently decrease inflammatory events has been focus of the current research.This perspective provides an overview of the development of MPO inhibitors, their mechanism of action and the review of molecules that were in clinical trials as promising MPO inhibitors.     

Dapsone for the treatment of doxorubicin extravasation injury in the rat

Doxorubicin is the most common antitumor drug implicated in serious extravasation injuries. Progressive tissue necrosis may lead to intense pain, chronic ulceration, and disfiguring tissue loss. This progressive necrosis is analogous to that seen with brown recluse spider bites, where dapsone is an established mode of therapy, minimizing the area of tissue loss by a proposed antiinflammatory mechanism. The backs of 50 Lewis rats were injected intradermally with 1 mg of doxorubicin in 1 cc of saline to simulate an extravasation injury. The rats were divided into five groups for treatment with oral dapsone 50 mg/kg/day: 10 were controls (no treatment), 10 were started the day before injury, 10 were started the day of injury, 10 were started the day after injury, and 10 were started 1 week after injury. The area of ulceration was calculated by planimetry. The data suggest that dapsone has little positive effect on healing extravasation ulcers.