In vitro activity of luromarin against tick-borne encephalitis virus

Luromarin, extracted from the sea alga Zostera asiatica, showed no cytotoxic activity when incubated with the SPEV cell culture and was characterized by virulicidal properties against highly pathogenic strain of TBE virus and the ability to inhibit the virus replication at the early stages of the infection.     

Effect of ethanol on activity of the plasma-membrane ATPase in, and accumulation of glycine by, Saccharomyces cerevisiae

The pH optimum of the ATPase activity in plasma membranes from Saccharomyces cerevisiae NCYC 431 from 8 h cultures was around 6.5 and that in membranes from organisms from 16 h cultures near 6.0. The Km[ATP] of the enzyme was virtually unaffected by the age of the culture from which organisms were harvested, although the Vmax of the enzyme in membranes from organisms from 8 h cultures was higher than that for organisms from 16 h cultures. Ethanol non-competitively inhibited ATPase activity in membranes, although the inhibition constant for the enzyme from organisms from 8 h cultures was lower than that from organisms from 16 h cultures. Glycine accumulation by the general amino acid permease was non-competitively inhibited by ethanol. Inhibition constants were virtually the same for glycine uptake by deenergized organisms from 8 h and 16 h cultures, but under energized conditions the value was greater for organisms from 16 h rather than 8 h cultures. The data indicate that inhibition of plasma-membrane ATPase activity by ethanol could account, at least in part, for inhibition of glycine accumulation by ethanol.     

5 new species of blackflies (Diptera: Simuliidae) from Siberia, the Far East and Central Asia

Five new species of the family Simuliidae Stegopterna byrrangii Yankovsky, sp. n. and Cnephia chaurensis Yankovsky sp. n., from Taimyr peninsula, Metacnephia olyutorii Yankovsky sp. n. from Kamchatka, Metacnephia karakechensis Yankovsky sp. n. and Montisimulium sonkulense Yankovsky sp. n. from Kirghizia, are described. Stegopterna byrrangii differs from related species: in male--from all known species of the genus by peculiar form of apical end of gonostyles, when apical spine is approximately parallel to inner surface of gonostyle; in female--from all known species of the genus by very long genital lobes (their length twice more than width), from S. longicoxa Rubzov, 1971 and S. duodecimata (Rubzov, 1940) by long stem of genital fork, absence of anterolateral sclerotized projections of branches of genital fork; in pupa--from all known species of the genus by number of filaments of respiratory organ (14-16, when in other species no more than 12). Cnephia chaurensis differs from related species: in male--from C. intermedia Rubzov, 1956 and C. toptchievi Yankovsky, 1996 by absence of medial projection or ridge of gonosternum, from C. intermedia by widened body of gonosternum, from C. toptchievi by ridge on gonocoxites and wide triangular projections near base of arms of gonosternum; in female--from C. intermedia by sclerotized finger-like anterolateral projections of branches of genital fork, subdivision of branches into two lobes, wide flat apical end of stem of genital fork. Metacnephia karakechensis differs from related species: in male--from M. subalpina (Rubzov, 1956) by form of gonostyles, absence of triangular medial projection of gonosternum, short widened gonofurca and more number of spines in parameres, from M. kirjanovae (Rubzov, 1956) and M. slepjani (Rubzov, 1967) by triangular body of gonosternum, widened gonostyles, more number of spines in parameres; in larva--from M. subalpina by more number of rays of primary fan of premandibles, less number of ridges of hooks in posterior attachment organ, from M. kirjanovae by short thick antennae; in pupa--from M. subalpina by character of divergence of upper thick filaments of respiratory organ, from M. kirjanovae by presence of thick filaments of the organ. Metacnephia olyutorii differs from related species: in female--from M. larunae Worobez, 1984 and M. aldanica Worobez, 1987 by thick sclerotized anterolateral projections of branches of genital fork, from M. taimyrica Patrusheva, 1976 by 1-ended (not 3-ended) anterolateral projections of branches of genital fork, large (not shortened) anal lobes, from M. crassifistula (Rubzov, 1956) by very long stem of genital fork. Montisimulium sonkulense differs from related species: in male--from M. brachystylum (Rubzov, 1976) by more prolonged gonostyles, less number of spines of parameres, from M. ocreastylum (Rubzov, 1956) by form of gonostyles, from M. odontostylum (Rubzov, 1947) by more number of spines of parameres; in female--from M. brachystylum and M. odontostylum by stricktly developed posteromedial projections of branches of genital fork, from species having posteromedical projections of branches of genital fork--from M. (ABSTRACT TRUNCATED)     

Metabolism of daidzein by intestinal bacteria from rhesus monkeys (Macaca mulatta)

PURPOSE: To identify the metabolites produced from an isoflavonoid, daidzein, by colonic bacteria of rhesus monkeys. METHODS: The metabolism of daidzein by the fecal bacteria of nine monkeys was investigated. Daidzein was incubated anaerobically with fecal bacteria, and the metabolites were analyzed by use of liquid chromatography and mass spectrometry. RESULTS: The fecal bacteria of all of the monkeys metabolized daidzein to various extents. Dihydrodaidzein was found in cultures of fecal bacteria from two monkeys; dihydrodaidzein and equol were found in cultures from four monkeys; dihydrodaidzein, equol, and an unknown metabolite (MW = 244) were found in cultures from one monkey; and dihydrodaidzein and the unknown metabolite were found in cultures from two monkeys. CONCLUSIONS: Similar to that in humans, variation was evident in the metabolism of isoflavonoids by fecal bacteria from rhesus monkeys. Some metabolites produced by fecal bacteria from monkeys were the same as those produced by fecal bacteria from humans.     

Temperature-induced excess mortality in Moscow,Russia

After considering the observed long-term trends in average monthly temperatures distribution in Moscow, the authors evaluated how acute mortality responded to changes in daily average, minimum and maximum temperatures throughout the year, and identified vulnerable population groups, by age and causes of death. A plot of the basic mortality–temperature relationship indicated that this relationship was V-shaped with the minimum around 18°C. Each 1°C increment of average daily temperature above 18°C resulted in an increase in deaths from all non-accidental causes by 2.8%, from coronary heart disease by 2.7%, from cerebrovascular diseases by 4.7%, and from respiratory diseases by 8.7%, with a lag of 0 or 1 day. Each 1°C drop of average daily temperature from +18°C to −10°C resulted in an increase in deaths from all non-accidental causes by 0.49%, from coronary heart disease by 0.57%, from cerebrovascular diseases by 0.78%, and from respiratory diseases by 1.5%, with lags of maximum association varying from 3 days for non-accidental mortality to 6 days for cerebrovascular mortality. In the age group 75+ years, corresponding risks were consistently higher by 13–30%. The authors also estimated the increase in non-accidental deaths against the variation of daily temperatures. For each 1°C increase of variation of temperature throughout the day, mortality increased by 0.3–1.9%, depending on other assumptions of the model.     

Hormonal stimulation of cyclic AMP accumulation and glycogen phosphorylase activity in calcium-depleted hepatocytes from euthyroid and hypothyroid rats.

Activation of glycogen phosphorylase by hormones was examined in hepatocytes isolated from euthyroid and hypothyroid female rats and incubated by Ca2+-free buffer containing 1 mM-EGTA. Basal glycogen phosphorylase activity was decreased in Ca2+-free buffer. However, the activation of hepatocyte glycogen phosphorylase, in the absence of extracellular Ca2+, in response to adrenaline, glucagon or phenylephrine was slightly lower, whereas that by vasopressin was abolished. The activation of glycogen phosphorylase by phenylephrine, adrenaline or isoproterenol (isoprenaline) in hepatocytes from euthyroid rats incubated in the absence of Ca2+ was not accompanied by any detectable increase in total cyclic AMP. The log-dose/response curves for activation of phosphorylase by phenylephrine or low concentrations of adrenaline were the same in hepatocytes from hypothyroid as compared wit euthyroid rats, whereas the response to isoproterenol was greater in hepatocytes from hypothyroid rats. However, the increases in total cyclic AMP accumulation caused by adrenaline or isoproterenol were greater in hepatocytes from hypothyroid rats than in hepatocytes from euthyroid rats. The increases in cyclic AMP accumulation caused by adrenaline or isoproterenol in Ca2+-depleted hepatocytes from hypothyroid rats were blocked by propranolol, a beta-adrenergic antagonist. In contrast, propranolol was only partially effective asan inhibitor of the activation of glycogen phosphorylase by phenylephrine or adrenaline in hepatocytes from hypothyroid rats and ineffective on phosphorylase activation in cells from euthyroid rats. These data indicate that the alpha-adrenergic activation of glycogen phosphorylase is not affected by the absence of extracellular Ca2+, and the extent to which total cyclic AMP was increased by adrenergic amines did not correlate with glycogen phosphorylase activation.     

Nonenzymatic isolation of Trichinella pseudospiralis infective L1 larvae at pH 7.4

Over half of the number of Trichinella pseudospiralis infective L1 larvae recovered from host carcasses by pepsin-HCl digestion were isolated from homogenized carcasses incubated in HBSS. More worms isolated by the latter method were viable compared to those isolated by pepsin-HCl digestion. When host carcasses infected with T. pseudospiralis were diced into pieces and incubated in HBSS, 30% more worms were recovered than from homogenized carcasses incubated in HBSS as above, and the majority of worms acquired by the former method were viable. The infectivity of T. pseudospiralis infective L1 larvae isolated from homogenized muscle in HBSS was 3.9 times greater than that for larvae recovered from homogenized carcasses by pepsin-HCl digestion. Only 4% and 0.8% of the number of T. spiralis recovered from homogenized muscle by pepsin-HCl digestion were isolated from homogenized or diced muscle incubated in HBSS, respectively. Fewer T. spiralis isolated from homogenized tissue in HBSS were viable compared to those recovered from homogenized carcasses digested in pepsin-HCl or diced carcasses incubated in HBSS.     

江淮丘陵地区油-稻轮作条件下土壤硫库变化研究

江淮丘陵地区下蜀系黄土母质发育的水稻土油2稻轮作试验表明, 油菜种植期间, 耕层土壤硫主要来源于耕层以下土层的补给, 其次是大气干湿沉降;该时期耕层土壤硫输出主要是油菜吸收, 其次是淋失.油菜种植期间耕层土壤硫输入量小于输出, 导致耕层土壤硫库下降8.76kg·hm^-2, 22%来自无机硫库的下降.水稻种植期间, 耕层土壤硫输入主要来自灌溉水, 其次是底土层的补给和大气干湿沉降;而硫输出主要是淋失, 其次是水稻吸收.耕层土壤硫输入量大于输出, 导致耕层土壤硫库增加18.69kg·hm^-2, 18%来自无机硫库的增加.全年油2稻轮作期间耕层土壤硫输入量大于输出, 导致耕层土壤硫库增加9.93kg·hm^-2, 13%来自无机硫库的增加.     

Proteomic analysis of secreted muscle components: search for factors involved in neuromuscular synapse formation

Denervated but not innervated skeletal muscles secrete polypeptides that are involved in neuromuscular synapse formation. With the aim of identifying such components, metabolically labeled polypeptides in extracts from denervated and innervated muscles were submitted to two-dimensional gel electrophoresis, and the abundance of individual molecular species was compared. Consistent differences between the proteomic maps from the two sources of muscles were seen. Likewise, proteomic maps of polypeptides from organ culture media conditioned by chronically denervated muscles and by control muscles revealed consistent differences, but the abundance of material within individual spots from conditioned media was not sufficient for analysis by mass spectrometry. Since it was not possible to match the patterns from muscle extracts and from conditioned media, it has been established that extract of Sol8 muscle cells was a satisfactory source of material for analysis. From 1,200 spots identified on the proteomic map from Sol8 cells by image analysis, some 140 have been defined by mass spectrometric analysis. In order to identify the components that are shared by secreted molecules from denervated muscles and Sol8 cells, a mixture of extracts from the two sources was co-electrophoresed and a shared proteomic pattern was established by visualization of metabolically labeled spots from the conditioned medium and of silver stained spots from the Sol8 cells. More than 100 spots sharing x/y coordinate localization could be seen on the pattern. Of these, fourteen were among those identified by mass spectrometry. It is concluded that co-electrophoresis of radioactively labeled polypeptides from conditioned media with extracts from Sol8 cells can be used to mark in the proteome of Sol8 cells those polypeptides that are secreted at low abundance by adult muscles. Their higher abundance in Sol8 cells opens the possibility for further scrutiny of spots by mass spectrometry or by microsequencing.     

Comparison of Cobb angles measured manually,calculated from 3-D spinal reconstruction,and estimated from torso asymmetry

While scoliotic spinal deformity is traditionally measured by the Cobb angle, we seek to estimate scoliosis severity from the torso surface without X-ray radiation. Here, we measured the Cobb angle in three ways: by protractor from postero-anterior X-ray, by computer from a 3-D digitized model of the vertebral body line, and by neural-network estimation from indices of torso surface asymmetry. The estimates of the Cobb angle by computer and by neural network were equally accurate in 153 records from 52 patients (standard deviation of 6 degrees from the Cobb angle, r=0.93), showing that torso asymmetry reliably predicted spinal deformity. Further improvements in predictive accuracy may require estimation of other 3-D indices of spinal deformity besides the Cobb angle with its wide measurement variability.     

Potential role of annexin AnnAt1 from Arabidopsis thaliana in pH-mediated cellular response to environmental stimuli

Plant annexins, Ca(2+)- and membrane-binding proteins, are probably implicated in the cellular response to stress resulting from acidification of cytosol. To understand how annexins can contribute to cellular ion homeostasis, we investigated the pH-induced changes in the structure and function of recombinant annexin AnnAt1 from Arabidopsis thaliana. The decrease of pH from 7.0 to 5.8 reduced the time of the formation of ion channels by AnnAt1 in artificial lipid membranes from 3.5 h to 15-20 min and increased their unitary conductance from 32 to 63 pS. These changes were accompanied by an increase in AnnAt1 hydrophobicity as revealed by hydrophobicity predictions, by an increase in fluorescence of 2-(p-toluidino)naphthalene-6-sulfonic acid (TNS) bound to AnnAt1 and fluorescence resonance energy transfer from AnnAt1 tryptophan residues to TNS. Concomitant lipid partition of AnnAt1 at acidic pH resulted in its partial protection from proteolytic digestion. Secondary structures of AnnAt1 determined by circular dichroism and infrared spectroscopy were also affected by lowering the pH from 7.2 to 5.2. These changes were characterized by an increase in beta-sheet content at the expense of alpha-helical structures, and were accompanied by reversible formation of AnnAt1 oligomers as probed by ultracentrifugation in a sucrose gradient. A further decrease of pH from 5.2 to 4.5 or lower led to the formation of irreversible aggregates and loss of AnnAt1 ionic conductance. Our findings suggest that AnnAt1 can sense changes of the pH milieu over the pH range from 7 to 5 and respond by changes in ion channel conductance, hydrophobicity, secondary structure of the protein and formation of oligomers. Further acidification irreversibly inactivated AnnAt1. We suggest that the pH-sensitive ion channel activity of AnnAt1 may play a role in intracellular ion homeostasis.     

Dependence of the structure of component parasite communities on host age

Dependence of the structure of component parasite communities on host age is studied by the example of parasite communities in minnow and grayling from the North Dvina and Pechora rivers basins. Parasite communities from immature host groups are revealed to be different from those in mature fishes by lesser number of parasite individuals and biomass, number of groups discriminated by the ratio of biomasses, and frequently by lesser number of species. Indices of diversity describing parasite communities from hosts of different ages are nearly always the same in the area examined. This observation needs in verification because values of the indices characterizing parasite communities from fishes of different ages are not equal in the parasite communities from grayling of different age from the Pinega river and minnow from the Chovju river (Dorovskikh, 2002).     

Relations between contractile responses and beta-adrenoceptors in gastric fundus of diabetic rats.

Contractile responses to norepinephrine (NE), and the population of beta-adrenoceptors, were determined in gastric fundus smooth muscle from rats with diabetes induced by streptozotocin (STZ), and age-matched controls. Relaxation and/or contraction of fundus strips of controls and diabetics were induced by 10(-5)M NE. Responses to NE were mainly relaxation in gastric fundus isolated from controls, and contraction in fundus isolated from diabetics. Contraction was blocked by 10(-8) M prazosin and relaxation was blocked by 10(-6) M propranolol. Relaxation by isoproterenol of contraction induced by 10(-6) M acetylcholine was significantly less in fundus from diabetics than in that from controls. The number of beta-adrenoceptors, measured with [125I] iodocyanopindolol as a ligand, was significantly less in gastric fundus membrane isolated from diabetics than in that from controls, but affinity was no different. The level of plasma catecholamine was higher in diabetics than in controls. Results suggest that depression of gastric fundus relaxation and increase of contraction by NE in diabetics could be due to fewer beta-adrenoceptor binding sites caused by down-regulation by higher catecholamine level in diabetic rats.     

研究荧光标记BCAA及AAA在大鼠体内代谢

使用DNS-Cl标记BCAA及AAA等六种氨基酸,观察这些氨基酸在大鼠小肠的吸收,血液的清除以及肝脏、肌肉、肾脏、脑等器官的摄取及排空情况。结果:小肠在30分钟开始吸收,4小时部分排空;血液在4小时开始清除,16小时清除完毕;肝脏、肌肉、肾脏、脑组织等,都在30分钟全部或部分摄取,但各器官对不同氨基酸,排空的时间不尽相同。     

Secretion and uptake of nascent hepatic very low density lipoprotein by perfused livers from fed and fasted rats

Livers from fed or 24-hr fasted male rats were perfused in a recycling system. VLDL labeled with [1-14C]oleate (95% in triglyceride), produced in separate perfusions of livers from fed rats, was added to the medium as a pulse. Uptake of VLDL 14C-labeled triglyceride by livers from fasted rats was less than that from fed rats regardless of addition of oleate. During the interval in which radioactive triglyceride was taken up, the mass of triglyceride in the medium increased, indicative of the synthesis and net secretion of triglycerides. The rates of secretion of VLDL and uptake of VLDL were both more rapid in livers from fed rats in comparison to those from fasted animals. It was calculated that about 50% of the triglyceride synthesized and secreted by the liver was taken back by livers from fed rats. The VLDL from livers of fasted rats did not contain any apoE detectable by SDS gel electrophoresis or by radioimmunoassay when no fatty acid or 166 mumol of oleic acid was infused. In contrast, apoE comprised 6% of the VLDL apoprotein derived from perfusion of livers from fed animals in the absence of added fatty acid, and 20% when the fed livers were infused with 166 mumol of oleic acid. However, the net output (accumulation) of apoE by fasted liver was only two-thirds that from fed livers. When lipoprotein-free rat plasma containing apoE (4 mg/dl) was used in place of bovine serum albumin, the VLDL secreted by livers from either fed or fasted rats contained apoE and was taken up to a similar extent by such livers. These data suggested that the apoE of the d greater than 1.21 g/ml fraction was transferred to newly secreted VLDL which then stimulated uptake of the VLDL by livers from fasted rats. With further stimulation of secretion of VLDL triglyceride by infusion of 332 mumol of oleic acid/hr, the percent of apoE in the VLDL secreted by livers from fasted rats increased to 20%, which was similar to that of the VLDL produced by livers from fed rats when either 166 or 332 mumol/hr was infused. These data suggest a relationship between rates of hepatic secretion of VLDL (TG) and apoE, and the association of apoE with the secreted VLDL. During fasting, reduced secretion of both VLDL and apoE resulted in a VLDL particle that was considerably diminished in content of apoE and, therefore, that would be taken up by the liver at a reduced rate, in comparison to that observed in the fed animal.     

Beta2 integrin modulates platelet caspase activation and life span in mice

We explored the role of CD18 (beta2 integrin) in platelet physiology, using mice genetically deficient in CD18 (CD18 -/-), or its main ligand CD54 (ICAM-1, CD54 -/-). CD18 and CD11a were evident in platelets from +/+, but not from CD18 -/- mice, as seen by immunofluorescence or Western blots. CD18 mRNA was also detectable by RT-PCR in platelets from +/+, but not from CD18 -/- mice. The life span of platelets was significantly shorter in CD18 -/- than in +/+ or CD54 -/- mice, as seen by in vivo biotinylation. When a local inflammation was elicited by the intra-tracheal injection of TNF, labeled platelets from +/+, but not from CD18 -/- donors, did localize in the lung. The content of Bcl-3 was about 20-fold higher in platelet from CD18 -/-, than in those from +/+ or CD54 -/- donors, as seen on Western blots or by immunofluorescence and flow cytometry, while the amount of pro-caspase-3 was decreased. An activation of caspases in platelets from CD18 -/- was also evidenced by protease assays. Accordingly, gelsolin, a protein cleaved by caspase-3, showed a low-molecular-weight band in platelets from CD18 -/- but not from +/+ donors. These results demonstrate that the beta2 integrin, present in mouse platelets, modulates caspase activation and consequently platelet life span and response to TNF.     

Generalist entomopathogens as biological indicators of deforestation and agricultural land use impacts on Waikato soils

The relative abundance of entomopathogenic nematodes and fungi was estimated for 10 sites in each of indigenous forest, pasture, and cropland habitats by baiting soil samples with Galleria larvae. The steinernematid Steinernema feltiae (Filip) was the dominant nematode, occurring in soils from all three habitat types. The heterorhabditid Heterorhabditis zelandica Poinar was recovered only from soils of podocarp (Dacrycarpus dacrydioides (A. Rich.)) forests. Galleria infection by nematodes was higher in soils from forest habitat than in soils from pasture and cropland. Among the sampled forests, nematode infection was higher in soils from podocarp stands than those from broadleaf stands. The deuteromycete fungi Beauveria bassiana (Balsamo) Vuillemin, Metarhizium anisopliae (Metschnikoff) Sorokin, and Paecilomyces cf. cicadae (Miquel) Samson, and a Entomophthorales zygomycete, tentatively identified as a Tarchium species, were recovered from the Galleria baits. Infection of Galleria by B. bassiana and M. anisopliae occurred in soils from all habitat types, while that by P. cicadae occurred only in soils from forest habitats. Tarchium was recovered from a single pasture site. The frequency of Galleria infection by these entomopathogenic fungi collectively, and by B. bassiana alone, was higher in pasture soils than in soils from either forest or cropland. These results are discussed in relation to disturbance effects of land use changes and the potential role of generalist entomopathogens as biological indicators of soil health.     

Regulation of class II MHC gene expression by macrophages from Bcgr and Bcgs mice

The presence of class II mRNA was determined following stimulation of macrophages from Bcgr and Bcgs mice with rIFN-gamma. Despite the continuous expression of surface I-A glycoprotein by macrophages from Bcgr mice, class II mRNA was no longer present. The transient expression of I-A by macrophages from Bcgs mice, however, was accompanied by the disappearance of class II mRNA from the cells. Restimulation of macrophages from Bcgs mice, with rIFN-gamma resulted in the reappearance of class II mRNA and surface I-A expression. The reappearance of class II mRNA and the surface expression of I-A glycoprotein was inhibited by PGE2. These results indicate that differences in I-A expression by macrophages from Bcgr and Bcgs are not at the level of class II gene expression.     

Comparison of Cobb Angles Measured Manually,Calculated from 3-D Spinal Reconstruction,and Estimated from Torso Asymmetry

While scoliotic spinal deformity is traditionally measured by the Cobb angle, we seek to estimate scoliosis severity from the torso surface without X-ray radiation. Here, we measured the Cobb angle in three ways: by protractor from postero-anterior X-ray, by computer from a 3-D digitized model of the vertebral body line, and by neural-network estimation from indices of torso surface asymmetry. The estimates of the Cobb angle by computer and by neural network were equally accurate in 153 records from 52 patients (standard deviation of 6° from the Cobb angle, r =0.93 ), showing that torso asymmetry reliably predicted spinal deformity. Further improvements in predictive accuracy may require estimation of other 3-D indices of spinal deformity besides the Cobb angle with its wide measurement variability.     

Chemiosmotic lysis and insulin secretion: studies of isolated granules, intact and permeabilised rat islets of Langerhans

The possible involvement of chemiosmotic lysis of secretory granules in the exocytosis of insulin from pancreatic beta cells was investigated by comparing insulin release from isolated secretory granules, from intact islets of Langerhans, and from electrically permeabilised islets. Lysis of isolated granules was stimulated by ATP in the presence of Mg2+. ATP-induced granule lysis was pH and temperature dependent and was inhibited by collapsing the pH gradient across the granule membrane by removal of permeant anions, or by increasing the extragranular osmolarity. However, insulin secretion from intact islets in response to glucose, a phosphodiesterase inhibitor or a Ca2+ ionophore was only partially inhibited by anion replacement, while Ca2+ -induced insulin release from electrically permeabilised islets was not affected by altering the extragranular or intragranular pH. These results suggest that studies of the stability of isolated granules in vitro do not necessarily relate to insulin release from whole cells, and do not support a major role for chemiosmotic lysis of secretory granules in the exocytotic release of insulin.