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TiO2 thin films were used as photocatalysts in CO2 photoconversion to solar fuels under UV irradiation. The films were deposited via sputtering while varying the working pressure for each deposition and could produce CH2O2 and CH3OH. According to Raman results, the films Ti-1 (3.87 Pa) and Ti-2 (3.73 Pa) deposited at high working pressure have a higher contribution of brookite phase compared with the films Ti-3 (3.47 Pa) and Ti-4 (3.33 Pa) obtained at low working pressure. Likewise, a higher presence of the brookite phase increases the atomic concentration of C–O bonds and −OH groups on the film surface, which reaches the CO2 photoconversion, as the TiO2 capacity to adsorb CO2 on the surface is determined by the O and C species present in the film. Therefore, the films Ti-1 and Ti-2, deposited at high working pressures and significantly contributing to the brookite phase, can produce 0.56 and 0.94 μmol of CH3OH, respectively. Meanwhile, according to the Raman and XPS results, the highest formic acid production was obtained from the Ti-3 and Ti-4 films, which show a less brookite phase presence and a higher contribution of O2− ions in TiO2. [ABSTRACT FROM AUTHOR]

Doping titanium dioxide has become a strategy for enhancing its properties and reducing its recombination issues, with the aim of increasing its efficiency in photocatalytic processes. In this context, this work studied its deposition over glass substrates using a sol–gel dip coating methodology. The effect of doping TiO2 with Zirconium cations in low molar concentrations (0.01, 0.05, 0.1%) in terms of its structural and optical properties was evaluated. The structural characterization confirmed the formation of amorphous thin films with Zr introduced into the TiO2 cell (confirmed by XPS characterization), in addition to increasing and defining the formed particles and their size slightly. These changes resulted in a decrease in the transmittance percentage and their energy band gap. Otherwise, their photocatalytic properties were evaluated in hydrogen production using ethanol as a sacrificial agent and UV irradiation. The hydrogen evolution increased as a function of the Zr doping, the sample with the largest Zr concentration (0.1% mol) being the most efficient, evolving 38.6 mmolcm−2 of this gas. Zr doping favored the formation of defects in TiO2, being responsible for this enhancement in photoactivity. [ABSTRACT FROM AUTHOR]

CO2 photoconversion to solar fuels requires materials with a high affinity to the acidic CO2, and MgO and Mg(OH)2 films represent good candidates due to their basic sites are highly active for CO2 capture in a wide interval of temperatures. However, the deposition of MgO and Mg(OH)2 as thin film is difficult to obtain by traditional methods. As an alternative, in this work, the successive ionic layer adsorption and reaction (SILAR) method is proposed to obtain MgO/Mg(OH)2 mixtures over glass substrates at significantly lower temperatures (200–400 °C). The films were tested as photocatalysts in the CO2 photoconversion to solar fuels (HCOOH and CH3OH) under UV–visible-NIR irradiation. The as-prepared films exhibited the hexagonal structure of the Mg(OH)2 phase. As the temperature increased, XRD and XPS analysis confirmed the presence of orthorhombic MgO, while the morphology remains with similar grains with an estimated size of 1 µm. The annealing temperatures change the chemical species (Mg–O, Mg–OH, and Mg–CO3) on the films affected their photocatalytic activity. The films exhibited high affinity for CO2 due to the presence of defects (F and F+ centers) in both phases. The photocatalytic behavior was directly related to OH− species present in each sample. According to the results, it seems that fewer hydroxides and defects on the films favored higher efficiencies for the CO2 photoconversion. In addition, the films were exposed to accelerated weathering tests to evaluate their efficiency for more extended periods. The results indicated that the aged films still have activity for CO2 photoconversion. [ABSTRACT FROM AUTHOR]

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Aim Delay in commencing adjuvant therapy for colorectal cancer seems to impair survival in some retrospective studies. This study was planned to evaluate its impact on survival. Methods This was a retrospective study enrolling patients registered from 2000 to 2012 in two large cancer-dedicated institutions in Brazil. The primary outcome was overall survival according to early vs late chemotherapy initiation. The interval between the primary surgery and the start of adjuvant chemotherapy was calculated. Survival was estimated using the Kaplan-Meier method and the impact of multiple prognostic factors on survival by Cox regression analysis. Results By the end of 2012, a total of 1963 Stage II and III colorectal patients were identified and 1318 patients received adjuvant chemotherapy, with 22% and 46% of those starting adjuvant chemotherapy within 6 weeks and 8 weeks of surgery. The median period of follow-up was 41 months. Patients starting chemotherapy within 6-8 weeks of surgery had longer overall survival compared with those who started after (6 weeks vs later, hazard ratio 0.76, 95% CI 0.57-0.99, P = 0.046; 8 weeks vs later, hazard ratio 0.74, 95% CI 0.59-0.93, P = 0.011). In the multivariate analysis, age, stage, histological grade, angiolymphatic invasion, emergency surgery and preoperative therapy were independent prognostic factors, but the interval between surgery and start of adjuvant therapy was not. Conclusion In this large retrospective study, the standard prognostic factors impacted on survival whereas the timing of adjuvant therapy did not. Patients with delayed adjuvant chemotherapy may have worse prognostic factors which could play a major role in their poor outcome. [ABSTRACT FROM AUTHOR]

In this work, a systematic variation of the processing parameters, such as gas mixture, time of nitriding and current density, has been carried out to determine how these will influence the sliding wear behaviour of plasma nitrided layers on AISI H13. Optical profilometry and scanning electron microscopy were used to characterise the worn sample and to determine the wear mechanism. The results revealed the strong influence of the current density on the formation of different diffusion and nitrided layers, which explained the wear behaviour of the studied samples, especially for those which were tested at room temperature. It was found that the wear volume of the nitrided samples without the existence of a compound layer was around 200 times smaller than that of the untreated samples and nearly 7 times smaller than that of nitrided samples when the compound layer was present. During the tests carried out at 300°C, the wear behaviour for the nitrided samples was worse than that for the untreated ones, due to the formation of higher amount of debris as a consequence of the breakdown of the compound layer and the subsequent removal of the enriched CrN from the ferritic matrix, generating a severe abrasion wear mechanism. All the samples tested at 600°C presented a similar tribological behaviour irrespective to the processing parameters values chosen for nitriding, as a consequence of the formation of a uniform and 'lubricious' iron oxide layer which, however, was insufficient to protect the steel against wear at this temperature to the levels which were obtained for the samples tested at room temperature. [ABSTRACT FROM AUTHOR]

In the present paper, five different operating conditions of a plasma nitriding process on annealed AISI H13 were investigated. A systematic variation of processing parameters such as gas mixture, time of processing and current density has been carried out in order to study their effect on microstructure. X-ray diffraction, microhardness testing and scanning electron microscopy techniques coupled with semiquantitative energy dispersive X-ray analysis were used to characterise the nitrided samples. The results revealed that the current density during plasma processing has a considerable influence on both the compound layer formation and depth of the diffusion zone. Also, it has been shown that the geometrical configuration of the workload may affect the uniformity of the diffusion zone, giving rise to the presence of heterogeneous nitrogen distribution zones along it. [ABSTRACT FROM AUTHOR]

In the present work, study on a systematic variation of the processing parameters such as gas mixture, time of nitriding and current density has been carried out in order to determine how these will influence the sliding wear behaviour of plasma nitrided layers on AISI H13. Optical profilometry and scanning electron microscopy were used to characterise the worn sample and to determine the wear mechanism. The results revealed a strong influence of the current density on the formation of different diffusion and nitrided layers, which explained the wear behaviour of the studied samples, especially for those tested at room temperature. It was found that the wear volume of the nitrided samples without the existence of a compound layer was ∼200 times smaller than the wear volume of the untreated samples and nearly seven times smaller than the wear volume of nitrided samples when the compound layer was present. When the tests were carried out at 300°C, the wear behaviour was worse for the nitrided samples than for the untreated ones, owing to the formation of higher amount of debris as a consequence of the breakdown of the compound layer and the subsequent removal of the enriched Cr–N from the ferritic matrix, generating a severe abrasion wear mechanism. All the samples tested at 600°C presented a similar tribological behaviour irrespective of the processing parameters values chosen for nitriding, as a consequence of the formation of a uniform and 'lubricious' iron oxide layer which, however, was insufficient to protect the steel against wear at this temperature to the levels which were obtained for the samples tested at room temperature. [ABSTRACT FROM AUTHOR]

ZnO thin films with grain morphology were obtained over glass substrates using the sol-gel method; these films were used as substrates to grow ZnO nanorods using chemical bath deposition (CBD) technique. The transmittance percentage of two films was over 85% and the electrical resistivity decreased when ZnO nanorods were grown over the ZnO thin film with grain morphology. A laser is incident at the thin film surface to measure the electrical response as a function of the wavelength change, it is found that electrical response is higher for the thin films with ZnO nanorods; these results shows that ZnO nanorods are a good choice for their use in photosensors. [ABSTRACT FROM AUTHOR]

TiO 2 thin films with mixtures of the anatase, rutile, and brookite phases were deposited on glass substrates via magnetron sputtering. Based on XRD and Raman results, the TiO 2 -0.47 and TiO 2 -3.47 films principally contained the brookite phase, while the TiO 2 -1.27 and TiO 2 -2.13 films were primarily anatase. The capacities of the TiO 2 films to adsorb heavy metals were tested with Cr(VI) and Fe(III) solutions, and the maximum Cr(VI) and Fe(III) adsorption capacities were realized with the TiO 2 -0.47 film (334.5 mg/g) and TiO 2 -3.47 film (271.3 mg/g), respectively. SEM‒EDS results revealed the presence of Cr and Fe on the surfaces of the films, thus corroborating the ability of the TiO 2 films to adsorb and remove heavy metals. They are strong candidates for use in wastewater treatment plants., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Luminescent properties of alexandrite crystals (BeAl 2 O 4 :Cr 3+ ) are studied. Partial readouts of thermoluminescence (TL) and optically stimulated luminescence (OSL) signals were obtained to infer the relations of OSL and TL trapping centers. Four TL peaks were studied; their intensities decrease at different rates by illumination with blue OSL light. The OSL curve shows two-time components (exponential decay constants 3.15 and 28.4 s). The short time component is preferentially diminished when the sample is heated to temperatures from 363 to 513 K. About 50% of the whole TL intensity remains after a complete 300s OSL readout., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Most Chagas patients belong to the chronic indeterminate stage, in which pharmacological treatment has an inconclusive outcome., Objective: To evaluate the efficacy of nifurtimox treatment in chronic asymptomatic Trypanosoma cruzi infection., Methods: We performed a systematic review and meta-analysis of electronically published literature, with no language, type of study, age or gender restrictions, until September 2010. Studies of chronic asymptomatic Chagas disease patients treated exclusively with nifurtimox were included in the analysis. Treatment efficacy was evaluated using parasitological or serological parameters., Results: Of 463 identified studies, 7 were finally selected: 6 observational studies and 1 randomized clinical trial; 4 of the studies were in adults, 3 in children < 14 years. In 6 studies, outcomes were defined by serological techniques. Summary estimate (log odds) was 0.37 (CI9 -1.32 - 2.07)., Conclusions: The analyzed studies gave discordant results. Those might be explained by differences in the populations studied, follow-up periods, diagnostic techniques, and sample size. More studies are necessary to obtain conclusive results about treatment efficacy of nifurtimox in this clinical phase of T. cruzi infection.

Fewer ethnic minorities, especially Asian-Americans, become organ donors. There are cultural, religious, and personal barriers to becoming a designated organ donor. Factors that promote or inhibit organ donation in Asians, especially Filipinos, are not well understood. We conducted a series of focus groups to identify barriers and facilitators to organ donation (deceased donor) among Filipinos. Six focus groups were conducted with church members, adolescents, nurses, physicians, organ recipients, and organ donor families. The mean age of adult participants (n = 57) was 52.3 +/- 15 years, 83% were Catholic, and 72% were female. A qualitative theme analysis methodology identified dominant themes related to organ donation in the participants. The major themes were: awareness of organ donation (38%), family beliefs (25%), religion/spirituality (10%), attitude/emotions (10%), personal experience with organ donation (8%), health profession (6%), and cultural issues (3%). Seventy-five percent of the comments about awareness reflected a positive awareness of cultural issues regarding organ donation, and the rest reflected a lack of awareness or misconceptions. Almost every theme was mentioned in all six focus groups. Understanding a specific ethnic group's knowledge, attitudes, and cultural beliefs regarding organ donation is important in the development of educational campaigns to encourage organ donation in ethnic minority populations.

This paper addresses the association between use of a herbicide and anecdotal reports of reduced dung degradation and dung beetle populations. Dung beetles were monthly collected at two adjacent ranches in Mexico. Ranches were similar in area, elevation, exposition, soil, and vegetation, but differed in weed control. Ranch A controlled weeds manually, and ranch B controlled unwanted vegetation with applications of the herbicide Tordon 101M. The main species recovered on each ranch (Ataenius apicalis) was significantly more abundant at ranch A than at ranch B. Conversely, similar numbers of a second species, Ataenius sculptor, were recovered from both ranches. Three lines of evidence support the tentative conclusion that herbicide applications may be causing a decline in populations of A. apicalis on ranch B. First, the greatest reductions of A. apicalis were observed during periods of herbicide application. Second, A. sculptor, apparently little affected by these same herbicide applications, is active primarily during months without herbicide applications. Third, preliminary results of laboratory studies show that exposure to herbicide can impair reproductive function of the dung beetle Canthon cyanellus.

Fertility ratio is defined here as the proportion of females that a male can impregnate after a constant period of in-polygyny living. This ratio was investigated in male rats after denervation of two pelvic floor muscles, the pubococcygeus and iliococcygeus. Denervation was carried out by transecting the somatomotor branch of the pelvic nerve. The lesion did not modify the sexual behavior of males or their overall fertility, but decreased the weight of the ejaculated seminal plug. Consequently, the number of days living in cohabitation to induce pregnancy was increased in lesioned males (approximately 13 days) compared with intact and sham animals (approximately 5 days). These results showed that the fertility ratio was optimal when intact/sham males cohabited with females for two consecutive estrous cycles, but that lesioned males needed up to four cycles to induce most pregnancies. Two hypotheses are raised by our results. The first is that pelvic floor denervation decreases the forceful tension required to expel the semen from the prostatic urethra to the vagina, then an incomplete seminal plug is expelled. The second is that denervation cut afferent fibers that reflexively promote the continence of the semen deposited in the prostatic urethra during seminal emission, allowing some to leak out before ejaculation. The latter hypothesis can also explain the recovery of the fertility ratio in lesioned males. It could be a compensatory mechanism mediated by the pudendal nerve supply to the coccygeus muscle, the other pelvic floor muscle.

Male rat motoneurons innervating the pubococcygeus muscle were located in the ventral nucleus of lamina IX at the sixth lumbar (L6) and first sacral (S1) spinal cord segments. Retrograde labeling with horseradish peroxidase-wheat germ agglutinin was transported up to second-order dendrites and revealed that these motoneurons have a "U-shaped arborization" of dendrites toward the intermediolateral and intermediomedial nuclei area of lamina VII. This dendritic organization makes a wide "final common path" that probably integrates afferent information from several sources, accounting for the participation of the pubococcygeus muscle in autonomic and somatic processes, such as those related to micturition and reproduction. Castration produced a decrement in the morphometry of these motoneurons. A main effect was a decrement in dendritic length. Steroid replacement indicated that testosterone and estradiol, but not dihydrotestosterone, are able to induce a recovery of morphometric alterations. However, estrogen induced recovery after 2 weeks of treatment, whereas testosterone took 4 weeks. Thus, it is proposed that supraspinal aromatization of testosterone in the male central nervous system might be an important process for the appropriate organization of the pubococcygeus muscle motoneurons and that estradiol seems to need a shorter time of action than testosterone because of differential up-regulation and down-regulation of steroid receptors.

Male rats exhibit erections in the presence of inaccessible estrous females, and we investigated which gonadal steroids regulate these noncontact erections (NCEs). Sexually experienced Wistar males (n >/= 8/group) were tested for NCE four times (every 3 days) before castration, after castration, and after receiving subcutaneous implants of 10-mm Silastic capsules that were empty or filled with crystalline testosterone propionate (TP), dihydrotestosterone (DHT), estradiol benzoate (EB), or DHT + EB (10 mm each). Before castration, males responded with NCE in approximately 50% of tests. No males had NCEs after castration, beginning 3 days after surgery. Also, no males responded after treatment with EB or empty capsules. After receiving implants of TP, DHT, or DHT + EB, 50% of males had NCEs, beginning with the first test 3 days after treatment. On every measure of NCE, males treated with DHT or DHT + EB were indistinguishable from each other and from TP-treated males. Among the sexual responses of male rats, NCE appears to be more sensitive than other behaviors to changes in gonadal condition. In its profile of response to gonadal steroids (testosterone+, dihydrotestosterone+, estradiol-), NCE is similar to reflexive erection, for which spinal systems are sufficient, and unlike copulation (T+, DHT-, E+), which depends on discrete areas of the brain. We nonetheless conclude that NCE depends on androgen-sensitive systems in the brain, but androgen-sensitive neurons in the lumbosacral spinal cord may also play a role., (Copyright 1999 Academic Press.)

We examined the effects in male rats of bilateral transection of two nerves previously implicated in erectile function, the viscerocutaneous branch of the pelvic nerve (Vc) and the hypogastric nerve (HgN). In Experiment 1 (conducted in Storrs), males underwent simultaneous or successive section of Vc and HgN and were tested for copulation, reflexive erection, and noncontact erection (NCE), i.e. in response to remote cues from estrous females. NCE is considered to be analogous to 'psychogenic' erection in humans, for which the HgN has been ascribed a significant role. In all three types of test, males had a moderate to severe deficit in erectile function after Vc transection. Section of HgN alone had no apparent pro- or anti-erectile effect in any context, nor did it affect the decrement resulting from Vc surgery. Regardless of treatment, all groups retained some erectile potential in each type of test. The loss of bladder function after Vc surgery and of seminal plug deposition after HgN section gave evidence that the targeted nerves were in fact severed. In Experiment 2 (conducted in Xalapa), males were tested only for NCE, but (a) they were tested every 3 days beginning 3 days after each surgery, (b) the interval between the two surgeries was more than 2 weeks, rather than 1 week as in Experiment 1, to allow more time for recovery from general effects of surgery and for hypothetical plasticity of neural function. In the first test after the first surgery, all groups had a modest reduction in the proportion of males displaying NCE, relative to sham-operated males. However, this deficit did not extend to measures of NCE latency or number, and was absent after the second test. After the second surgery, when all males except those with sham operations had both nerves cut, none of the groups exhibited a significant deficit in NCE, and all groups had at least one test in which at least half the males responded. Thus, (a) HgN section did not significantly impair NCE, reflexive erection, or copulation; (b) Vc section impaired, but did not eliminate, erection in all three contexts, but even those effects may be transient; and (c) transection of both nerves, simultaneously or successively, did not cause a greater impairment in erection than did cutting just the Vc. We infer that the HgN may have no pro-erectile role in erection in rats, even in a model analogous to psychogenic erection. The Vc is probably the most important nerve mediating pro-erectile function in NCE, as in reflexive erection and copulation, but this nerve may not be essential for erection in rats in any context, at least in some males.

alpha 1-Antitrypsin (alpha 1-AT) and alpha 1-antichymotrypsin (alpha 1-ACHY) are closely related protease inhibitors, synthesized primarily by the liver, which play major roles in modulation of the inflammatory response. Previously, we had shown that MCF-7 human breast cancer cells were able to synthesize active alpha 1-AT and alpha 1-ACHY and that the synthesis of both inhibitors varied among different MCF-7 sublines. We now show that when MCF-7(ML) cells (a subline synthesizing low levels of alpha 1-AT) are grown in soft agar in medium depleted of its trypsin inhibitory capacity (i.e. alpha 1-AT-free), addition of alpha 1-AT (50 micrograms/ml) significantly reduces colony formation in both the presence and absence of estradiol (34% and 44%, respectively). Under these conditions, incubation with 10(-7) M estradiol alone increased colony formation 2- to 3-fold. Colony formation was also significantly reduced by serum leukocyte protease inhibitor, which, like alpha 1-AT, is a potent inhibitor of elastase-like enzymes. We also found that a variety of inflammatory mediators, cytokines, and steroid hormones are able to stimulate synthesis of alpha 1-AT and alpha 1-ACHY by MCF-7 cells. Stimulation by interleukin-6 (IL-6; 200 U/ml), epidermal growth factor (4 nM), and estradiol (10(-7) M) was 2- to 3-fold, whereas stimulations by tetradecanoylphorbol-13-acetate (TPA; 80 nM) and IL-1 (10 U/ml) were 2- to 5-fold and 5- to 10-fold, respectively. In each instance, protein synthesis, monitored by immunoprecipitation of 35S-labeled proteins followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and steady state mRNA levels, monitored by Northern blot analysis with specific cDNA probes, increased to the same extent. Consistent with their ability to stimulate alpha 1-AT synthesis, TPA and IL-1 reduced colony formation in the absence of estradiol by 65% and 63%, respectively. In addition, the effects of both TPA and IL-1 could be reversed by antibody to alpha 1-AT. These results suggest that local synthesis of alpha 1-AT and possibly other protease inhibitors may be important in regulating the tumorigenic potential of MCF-7 breast cancer cells.

alpha 1-Antichymotrypsin (alpha 1-ACHY) and alpha 1-antitrypsin (alpha 1-AT) are closely related glycoprotein protease inhibitors, present in plasma and other extracellular fluids, that neutralize proteases released by leukocytes in response to trauma and inflammatory stimuli. Both inhibitors are synthesized primarily by hepatocytes, although lower levels of synthesis by monocytes and breast and intestinal epithelial cells have been demonstrated. Recently, the immunohistochemical localization of alpha 1-AT and alpha 1-ACHY in intrauterine and extrauterine human trophoblastic tissue has been reported. In the present study, we have sought to determine whether human trophoblast is also able to synthesize alpha 1-AT and alpha 1-ACHY. Messenger RNA for both inhibitors was found by Northern blotting in chorionic villi obtained from first trimester and term placenta. Substantial differences in messenger levels for both inhibitors among individual placentas were noted. alpha 1-ACHY and alpha 1-AT messenger was also present in trophoblast cells in primary culture. Synthesis of alpha 1-AT and alpha 1-ACHY protein was demonstrated by SDS-PAGE after immunoprecipitation of [35S]-labeled alpha 1-AT and alpha 1-ACHY from conditioned media of trophoblast cells in culture metabolically labeled with [35S]-methionine. It is of some interest that the M(r) of the alpha 1-AT and alpha 1-ACHY secreted by trophoblast were 50,000 and 49,000, respectively, compared with 54,000 and 68,000 for these proteins in plasma (or secreted by HepG2 human hepatoma and MCF-7 human breast cancer cells).(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of phospho-glycoprotein (HMW-GP), obtained from human platelet plasma membranes, on the aggregation and secretion of human platelets was studied. Incubation of PRP with 4 to 16 micrograms/ml of HMW-GP results in inhibition of ADP-, Epinephrine-, Collagen-, and Thrombin-induced platelet aggregation. The effect is mainly reflected on the secondary wave of aggregation. The inhibitory effect is partially overcome by higher concentrations of inducers, however, even under these conditions, a clear tendency towards disaggregation is observed. 5HT release (Col-induced) is strongly decreased from 50% to 4.5. The inhibitory effect on Thrombin-induced aggregation is markedly dependent on external calcium, being maximal at 5 mM calcium. The HMW-GP does not bind ADP or Thrombin. Membrane conformation is markedly affected, as evidenced by the effect of HMW-GP on the iodination of surface polypeptides of intact platelets. It is suggested that interaction of HMW-GP with the platelet membrane blocks the signal(s) transmission that links stimulus to activation. The inhibition observed might just represent an experimental amplification of the endogenous modulatory function that has been proposed for this high molecular weight phosphoglycoprotein.

We studied the effect of the methanol extract of garlic bulbs (EOG) and of three pure components isolated from it (F1, F2, F3), on human platelet aggregation induced by ADP, epinephrine, collagen, thrombin, arachidonate, PAF, and the ionophore A-23187. Incubation of PRP with EOG, either in methanol or in homologous PPP, inhibits platelet aggregation induced by all of the above mentioned agonists. F1, F2, and F3 also inhibit platelet aggregation, however, F3 was about four times more potent. Addition of EOG or F3 to platelets that have already been irreversibly aggregated by 10 microM ADP, induces rapid deaggregation. Inhibition of aggregation was still present after three hours. The inhibitory effect persisted even after the treated platelets were Gel-Filtered (GFP) or separated from plasma through a metrizamide gradient and resuspended in new homologous PPP. Thrombin-induced release of ATP from GFP was inhibited by 75-80% after EOG or F3 treatment. Incorporation of [3-H]-arachidonate by intact platelets was decreased by 50-60% in treated platelets. However, platelets incubated with the inhibitors after incorporation of radiolabeled arachidonate, although did not aggregate, produced, after thrombin activation similar amounts of radiolabeled TXB2 and lipoxygenase products as the controls. Electron microscopy of inhibited platelets, in the presence of thrombin, showed no degranulation but an increase of spherical forms. Our results suggest that the effects described might be mediate by a perturbation of the physicochemical properties of the plasma membrane rather than by affecting arachidonate or calcium metabolism in the cells. Chemical structures of F1, F2 and F3 have been provisionally assigned: F1 is diallytrisulfide, F2 is 2-vinyl-1,3-dithiene, and F3 is most probably allyl 1,5-hexadienyltrisulfide.

Functional and biochemical studies of platelets from human Chédiak-Higashi syndrome (CHS) are scarce and/or incomplete. In the present report, the aggregation response to a variety of inducers of platelet aggregation, the content of the dense granule constituents ATP, ADP, serotonin and calcium, the secretion of ATP, ADP, and calcium induced by thrombin, the total content of magnesium, the incorporation of 14C-adenine in the cytoplasmic pool of adenine nucleotides, as well as the content of intracellular cyclic-AMP, have been quantitated in six patients with CHS. Furthermore, data is presented on the kinetics of uptake of radiolabelled serotonin and its storage in human CHS platelets. An abnormal aggregation behaviour was found in all patients. However, the response of CHS platelets to the different inducers studied did not show a uniform pattern. The total content and the maximal amounts of the dense granule constituents secretable by thrombin were greatly decreased in all six patients. Total magnesium content was similar to that of normal platelets. The ATP/ADP ratio was higher than in controls. Uptake of radiolabelled serotonin by CHS platelets closely followed the uptake by normal platelets; during the first 2-3 min, however, incorporation of the amine by CHS platelets came rapidly to a plateau which contrasts with the steady, linear increase in uptake found in controls. CHS platelets loaded with radiolabelled serotonin and gel-filtered, showed a spontaneous release of radioactivity not observed in normal platelets under the same conditions. The cyclic-AMP content of CHS platelets was similar to that of normals. In contrast to platelets from patients with storage pool disease, the secretable calcium from CHS platelets represents a 67% of total platelet calcium (61% in normals), suggesting that the absolute values for the non-secretable portion in CHS platelets must be very low. The results reported confirm the existence of a true storage pool deficiency of the dense granule constituents as a common defect in CHS platelets. The variety of responses among patients, to the different aggregatory stimuli studied, can not be solely ascribed to the storage pool deficiency described.

Studies on a purified phospholipase A2 (PLA2) from human platelets show that the enzyme, which is copurified with the plasma membrane fraction, has a MW of approximately 50 K Dalton, requires Ca++, and has a pH optimum of 9.4. Under optimal conditions, PLA2 activity corresponds to at least 13 nmol/min/10(9) platelets. Unsaturated PL are preferred substrates and the enzyme is considerably more active on the aggregated form of the substrate than on the monomers. The specific activity is markedly affected by the quality of the interface, showing variations of more than 10-fold between different substrate forms. In the absence of detergents, a 4-fold increase in rate is observed when both products are present. Maximal rates are obtained at 20 mole percent of products to substrate. 1,2-Diglyceride and phosphatidic acid stimulate the hydrolysis of PC by the purified enzyme, however, in these forms of the substrate, neither of them are hydrolyzed. Activation of this enzyme by some intermediate of the phospholipase C pathway might play a role in the stimulus-linked release of platelet arachidonic acid.