Human cytomegalovirus seropositivity is associated with reduced patient survival during sepsis.

BACKGROUND: Sepsis is one of the leading causes of death. Treatment attempts targeting the immune response regularly fail in clinical trials. As HCMV latency can modulate the immune response and changes the immune cell composition, we hypothesized that HCMV serostatus affects mortality in sepsis patients. METHODS: We determined the HCMV serostatus (i.e., latency) of 410 prospectively enrolled patients of the multicenter SepsisDataNet.NRW study. Patients were recruited according to the SEPSIS-3 criteria and clinical data were recorded in an observational approach. We quantified 13 cytokines at Days 1, 4, and 8 after enrollment. Proteomics data were analyzed from the plasma samples of 171 patients. RESULTS: The 30-day mortality was higher in HCMV-seropositive patients than in seronegative sepsis patients (38% vs. 25%, respectively; p = 0.008; HR, 1.656; 95% CI 1.135-2.417). This effect was observed independent of age (p = 0.010; HR, 1.673; 95% CI 1.131-2.477). The predictive value on the outcome of the increased concentrations of IL-6 was present only in the seropositive cohort (30-day mortality, 63% vs. 24%; HR 3.250; 95% CI 2.075-5.090; p < 0.001) with no significant differences in serum concentrations of IL-6 between the two groups. Procalcitonin and IL-10 exhibited the same behavior and were predictive of the outcome only in HCMV-seropositive patients. CONCLUSION: We suggest that the predictive value of inflammation-associated biomarkers should be re-evaluated with regard to the HCMV serostatus. Targeting HCMV latency might open a new approach to selecting suitable patients for individualized treatment in sepsis.

Measurements of Strong-Interaction Effects in Kaonic-Helium Isotopes at Sub-eV Precision with X-Ray Microcalorimeters.

We have measured the 3d→2p transition x rays of kaonic ^{3}He and ^{4}He atoms using superconducting transition-edge-sensor microcalorimeters with an energy resolution better than 6 eV (FWHM). We determined the energies to be 6224.5±0.4(stat)±0.2(syst) eV and 6463.7±0.3(stat)±0.1(syst) eV, and widths to be 2.5±1.0(stat)±0.4(syst) eV and 1.0±0.6(stat)±0.3(stat) eV, for kaonic ^{3}He and ^{4}He, respectively. These values are nearly 10 times more precise than in previous measurements. Our results exclude the large strong-interaction shifts and widths that are suggested by a coupled-channel approach and agree with calculations based on optical-potential models.

Association between FTO, MC4R, SLC30A8, and KCNQ1 gene variants and type 2 diabetes in Saudi population.

Recent genome wide association studies identified many loci in several genes that have been consistently associated with type 2 diabetes mellitus in various ethnic populations. Among the genes that were most strongly associated with diabetes were fat mass- and obesity-associated, melanocortin 4 receptor, solute carrier family 30 member 8 (SLC30A8), and a member of the potassium voltage-gated channels. In the present study, we examined the association between variants in fat mass- and obesity-associated [rs9939609 (A/T)], melanocortin 4 receptor [rs17782313 (C/T), and rs12970134 (A/G)], SLC30A8 [rs13266634 (C/T)], and a member of the potassium voltage-gated channels [rs2237892(C/T)] genes in diabetes patients from Saudi Arabia. Genotypes were determined using the TaqMan single-nucleotide polymorphism genotype analysis technique. Minor allele frequency of the 4 variants tested was comparable between type 2 diabetes cases and controls. We observed an association between allele variants of SLC30A8 [rs13266634 (C/T)] and type 2-diabetes (P = 0.04). The other single-nucleotide polymorphisms examined in this study showed moderate or no correlation with diabetes in Saudis. Our data indicate that the SLC30A8 polymorphisms are associated with type 2 diabetes in the Saudi population. There is no evidence supporting an association between variants in the fat mass- and obesity-associated and melanocortin 4 receptor, and a member of the potassium voltage-gated channels genes and type 2 diabetes in the Saudi population.

Preliminary study of kaonic deuterium X-rays by the SIDDHARTA experiment at DAΦNE.

The study of the [Formula: see text] system at very low energies plays a key role for the understanding of the strong interaction between hadrons in the strangeness sector. At the DAΦNE electron-positron collider of Laboratori Nazionali di Frascati we studied kaonic atoms with [Formula: see text] and [Formula: see text], taking advantage of the low-energy charged kaons from Φ-mesons decaying nearly at rest. The SIDDHARTA experiment used X-ray spectroscopy of the kaonic atoms to determine the transition yields and the strong interaction induced shift and width of the lowest experimentally accessible level (1s for H and D and 2p for He). Shift and width are connected to the real and imaginary part of the scattering length. To disentangle the isospin dependent scattering lengths of the antikaon-nucleon interaction, measurements of [Formula: see text] and of [Formula: see text] are needed. We report here on an exploratory deuterium measurement, from which a limit for the yield of the K-series transitions was derived: [Formula: see text] and [Formula: see text] (CL 90%). Also, the upcoming SIDDHARTA-2 kaonic deuterium experiment is introduced.

Strong-interaction shifts and widths of kaonic helium isotopes.

The kaonic 3He and 4He [Formula: see text] transitions in gaseous targets were observed by the SIDDHARTA experiment. The X-ray energies of these transitions were measured with large-area silicon-drift detectors using the timing information of the [Formula: see text] pairs produced by the DAΦNE [Formula: see text] collider. The strong-interaction shifts and widths both of the kaonic 3He and 4He 2p states were determined, which are much smaller than the results obtained by the previous experiments. The "kaonic helium puzzle" (a discrepancy between theory and experiment) was now resolved.

Radiographers' perception of patient safety culture in radiology.

INTRODUCTION: Radiographers play a central role in patient safety because of their knowledge of and responsibilities in relation to the imaging process. To maintain safe care, the workplace must create a safety culture that enables sustainable safety work. AIM: This study aims to describe radiographers' perceptions of the patient safety culture in radiology units in Sweden. METHODS: The Swedish Hospital Survey of Patients' Safety Culture (S-HSOPSC) was used to gather descriptive data from 171 Swedish registered radiographers working in five radiology clinics distributed across 15 units. Fifty-one questionnaire items and one open-ended question were analysed, comprising perceptions of the overall safety grade, the frequency of number of reported risks and events, and 14 composites regarding patient safety dimensions. RESULTS: The radiographers' concerns surrounding the patient safety culture in their workplaces related to weaknesses regarding the safety dimensions "Staffing", "Frequency of error reporting", "Organizational learning - continuous improvement" and "Executive management support for patient safety". They perceived "Teamwork within the unit" to be a strength. CONCLUSION: Despite some weaknesses in the patient safety culture, the radiographers perceived that the overall patient safety level was good, in part because of their ability to spot risks in time. The executive management, however, needed to improve their feedback on safety measures; and another reason for some weaknesses in the patient safety culture could be staffing issues such as lack of time for meetings for continuous improvement. Managers and leaders have a great responsibility to establish a patient safety culture through support and good leadership. IMPLICATIONS FOR PRACTICE: An understanding of what creates a safety culture is important to prevent patient safety incidents.

Measurements of the strong-interaction widths of the kaonic (3)He and (4)He 2p levels.

The kaonic (3)He and (4)He X-rays emitted in the [Formula: see text] transitions were measured in the SIDDHARTA experiment. The widths of the kaonic (3)He and (4)He 2p states were determined to be [Formula: see text], and [Formula: see text], respectively. Both results are consistent with the theoretical predictions. The width of kaonic (4)He is much smaller than the value of [Formula: see text] determined by the experiments performed in the 70's and 80's, while the width of kaonic (3)He was determined for the first time.

First measurement of kaonic helium-3 X-rays.

The first observation of the kaonic (3)He 3d→2p transition was made, using slow K- mesons stopped in a gaseous (3)He target. The kaonic atom X-rays were detected with large-area silicon drift detectors using the timing information of the K+K- pairs of ϕ-meson decays produced by the DAΦNE e+e- collider. The strong interaction shift of the kaonic (3)He 2p state was determined to be -2±2(stat)±4(syst) eV.

Interleukin 17A and F and asthma in Saudi Arabia: gene polymorphisms and protein levels.

BACKGROUND: Asthma is a multifactorial disorder, and both genetic and environmental factors contribute to its development. We investigated the possible association between asthma and 5 single-nucleotide polymorphisms (SNPs) in the interleukin 17 (IL17) gene--rs17880588 (G/A) and rs17878530 (C/T) in IL17A and rs763780 (T/C), rs11465553 (T/C), and rs2397084 (G/A) in IL17F--and compared levels of the proteins IL17A and IL17F in asthma patients with those of controls. PATIENTS AND METHODS: The study group included 100 asthma patients and 102 ethnically matched controls. Genotyping was performed on purified DNA using reverse transcriptase-polymerase chain reaction with specific primers and probes. Levels of IL17A and IL17F were measured in plasma using enzyme-linked immunosorbent assay. RESULTS: Genotyping showed that AG heterozygotes of rs17880588 in IL17A were significantly more common in the control group than among the asthma patients (P < .05); no significant associations were observed for any of the other SNPs examined. Levels of IL17A and IL17F were both higher in asthma patients (IL17A, 2.242 [0.099] vs 2.752 [0.287] pg/mL; IL17F, 236.01 [38.28] vs 700 [201.078] pg/mL). The difference was statistically significant for IL17F (P = .025, t test). Levels of IL17A and IL17F were positively and significantly correlated in the asthma patients CONCLUSION: Of all the SNPs analyzed, only rs17880588 showed a significant association with asthma in the Saudi population we studied. Levels of IL17A and IL17F were significantly upregulated in the asthma patients. The morphology of IL17F appeared to affect expression levels.

Radiology departmental policy compliance with Swedish guidelines regarding post-contrast acute kidney injury for examinations with iodinated contrast media.

INTRODUCTION: Guidelines concerning intravenous iodinated contrast media (CM) during computed tomography (CT) examinations are important to follow to minimize the risk for post-contrast acute kidney injury (PC-AKI). The purpose of this study was to investigate the radiology departmental policy compliance with Swedish guidelines concerning PC-AKI. METHODS: In February 2020, an electronic survey was distributed to the responsible radiographer at 41 radiology departments in all university hospitals and medium-sized hospitals in Sweden. The questions focused on routines around renal functional tests, individualized contrast administration and handling of patients with diabetes mellitus taking metformin. RESULTS: The response rate was 83%. Seventy-six percent (n = 26) of radiology departments calculated estimated glomerular filtration rate (eGFR) from serum creatinine prior to CM administration, but only 24% (n = 8) followed the recommendation to calculate eGFR from both serum creatinine and cystatin C. For acute/inpatients, 55% (n = 18) followed the recommendation that renal functional tests should be performed within 12 h before CM administration. For elective patients, 97% (n = 33) followed the recommendation to have eGFR newer than three months which is acceptable for patients with no history of disease that may have affected renal function. Approximately 80% of the radiology departments followed the recommendation that CM dose always should be individually adjusted to patient eGFR. Seventy-six percent (n = 26) followed the recommendation to continue with metformin at eGFR ≥ 45 ml/min. CONCLUSION: Compliance with the national guidelines was high regarding routines around renal functional tests, dose adjustment of CM and metformin discontinuation. Improvements can be made in using both cystatin C and serum creatinine for eGFR calculations as well as ensuring renal function tests within 12 h for acute/inpatients with acute disease that may affect renal function. IMPLICATIONS FOR PRACTICE: This study raises awareness of the importance of adhering to guidelines in healthcare. To have knowledge about the current level of compliance regarding PCI-AKI is important to maintain and develop effective clinical implementation of guidelines. The variation in practice seen in this study emphasizes the need of more effective implementation strategies to ensure adherence with best practice.

Experimental search for the violation of Pauli exclusion principle: VIP-2 Collaboration.

The VIolation of Pauli exclusion principle -2 experiment, or VIP-2 experiment, at the Laboratori Nazionali del Gran Sasso searches for X-rays from copper atomic transitions that are prohibited by the Pauli exclusion principle. Candidate direct violation events come from the transition of a 2p electron to the ground state that is already occupied by two electrons. From the first data taking campaign in 2016 of VIP-2 experiment, we determined a best upper limit of [Formula: see text] for the probability that such a violation exists. Significant improvement in the control of the experimental systematics was also achieved, although not explicitly reflected in the improved upper limit. By introducing a simultaneous spectral fit of the signal and background data in the analysis, we succeeded in taking into account systematic errors that could not be evaluated previously in this type of measurements.

High-affinity binding of NADPH to camel lens zeta-crystallin.

Fluorescence spectrum of camel lens zeta-crystallin, a major protein in the lens of camelids and histicomorph rodents, showed maximum emission at 315 nm. This emission maximum is blue shifted compared to most proteins, including alpha-crystallin, and appeared to be due to tryptophan in highly hydrophobic environment. Interaction of NADPH with zeta-crystallin quenched the protein fluorescence and enhanced the fluorescence of bound NADPH. Analysis of fluorescence quenching suggested high-affinity interaction between NADPH and zeta-crystallin with an apparent Km<0.45 microM. This value is at least an order of magnitude lower than that suggested by activity measurements. Analysis of NADPH fluorescence showed a biphasic curve representing fluorescence of free- and bound-NADPH. The intersection between free- and bound-NADPH closely paralleled the enzyme concentration, suggesting one mole of NADPH was bound per subunit of the enzyme. Phenanthrenequinone (PQ), the substrate of zeta-crystallin, also was able to quench the fluorescence of zeta-crystallin, albeit weaker than NADPH. Quantitative analysis suggested that zeta-crystallin had low affinity for PQ in the absence of NADPH, and PQ binding induced significant conformational changes in zeta-crystallin.

Hydrophobicity of the NADPH binding domain of camel lens zeta-crystallin.

Interaction of camel lens zeta-crystallin with the hydrophobic probe 1-anilinonaphthalene-8-sulfonic acid (ANS) enhanced the ANS fluorescence and quenched the protein fluorescence. Both of these events were concentration-dependent and showed typical saturation curves suggesting specific ANS-zeta-crystallin binding. Quantitative analysis indicated that 1 mole zeta-crystallin bound at most 1 mole ANS. NADPH but not 9,10-phenanthrenequinone (PQ) was able to displace zeta-crystallin-bound ANS. These results suggested the presence of a hydrophobic domain in zeta-crystallin, possibly at the NADPH binding site. alpha-Crystallin as well as NADPH protected zeta-crystallin against thermal inactivation suggesting the importance of this site for enzyme stability. The NADPH:quinone oxidoreductase activity of zeta-crystallin was inhibited by ANS with NADPH as electron donor and PQ as electron acceptor. Lineweaver-Burk plots indicated mixed-type inhibition with respect to NADPH, with a K(i) of 2.3 microM. Secondary plots of inhibition with respect to NADPH indicated a dissociation constant (K'I) of 12 microM for the zeta-crystallin-NADPH-ANS complex. The K(i) being smaller than K'I suggested that competitive inhibition at the NADPH binding site was predominant over non-competitive inhibition. Like ANS-zeta-crystallin binding, inhibition was dependent on ANS concentration but independent of incubation time.

Phosphorylation of troponin I by protein kinase C: mechanism of inhibition by calmodulin and troponin C.

The mechanism by which calmodulin and troponin C influence phosphorylation of troponin I (TnI) by protein kinase C was investigated. The phosphorylation of TnI by protein kinase C requires the presence of acidic phospholipid, calcium and diacylglycerol. Light scattering intensity and fluorescence intensity experiments showed that TnI associated with the phospholipid membranes and caused extensive aggregation. In the presence of Ca2+, TnI-phospholipid interactions were prevented by approximately stoichiometric amounts of either troponin C or calmodulin. Troponin C was shown to completely inhibit phosphorylation of TnI by either protein kinase C or by phosphorylase b kinase. In contrast, calmodulin completely inhibited phosphorylation of TnI by protein kinase C, but had only little effect on TnI phosphorylation by phosphorylase b kinase. Inhibition by calmodulin did not appear to be due to interaction with PKC, since calmodulin mildly increased protein kinase C phosphorylation of histone III-S. The ratio of phosphoserine to phosphothreonine in protein kinase C-phosphorylated TnI remained approximately constant for reactions inhibited by up to 90% by calmodulin. TnI interactions with phospholipid and phosphorylation of TnI by PKC were also prevented by high salt concentrations. However, salt concentrations adequate to inhibit phosphorylation were sufficient to dissociate only TnI, but not protein kinase C from the membrane. These results suggest that the binding of TnI to phospholipid is required for phosphorylation by protein kinase C and that prevention of this binding by any means completely inhibited phosphorylation of TnI by protein kinase C.

Protein kinase C and annexins: unusual calcium response elements in the cell.

Protein kinase C and the annexins appear to share some unusual and potentially important membrane- and calcium-binding properties. While these proteins are calcium response elements, they are not calcium-binding proteins in the formal sense; at intracellular calcium concentrations, they only bind significant amounts of calcium when membranes or other suitable surfaces are present. The number of calcium ions bound per protein is large (> 8) and this stoichiometry, at the protein-membrane interface, may provide the large number of contact points needed for the very high-affinity interaction that is observed. The further ability of annexins and PKC to form structures with properties of integral membrane proteins may be important to provide a type of long-term cell signalling that produces a constitutively active kinase or ion channel activity. Selectivity for phospholipids in bilayer form is modest with respect to the acidic phospholipids but there is a surprising preference for phosphatidylethanolamine as the neutral phospholipid matrix. Along with other unusual properties, these proteins offer the potential for unique types of cell regulation events.

Adjunctive treatment with propylthiouracil or iodine following radioiodine therapy for Graves' disease.

The effect of radioiodine therapy in Graves' disease is gradual in onset and the subjects continue to be hyperthyroid for several weeks after such therapy. In a prospective study of 112 patients, we compared the usefulness of two commonly employed antithyroid drug regimens in controlling hyperthyroidism in the period immediately following radioiodine therapy. They received propylthiouracil (PTU, 100 mg orally three times a day), saturated solution of potassium iodide (10 drops once a day) or no drugs starting 5 days after radioiodine therapy. Thyroid status was monitored clinically and by serum thyroxine index and TSH measured at 4-6 week intervals over a 6-month period. The control or drug-treated groups did not differ in thyroid status 6 weeks after radioiodine. The PTU-treated group had greater incidence of hyperthyroidism and a lower incidence of hypothyroidism at 6 months. However, the differences were explained on the basis of a greater incidence of large goiters that appeared to confer relative radioresistance in the PTU group. We conclude that patients with mild to moderate hyperthyroidism do not benefit from adjunctive treatment with PTU or potassium iodide immediately after radioiodine therapy.

Interaction of camel lens zeta-crystallin with quinones: portrait of a substrate by fluorescence spectroscopy.

Interaction of camel lens zeta-crystallin, an NADPH:quinone oxidoreductase, with several quinone derivatives was examined by fluorescence spectroscopy and activity measurements. Fluorescence of zeta-crystallin was quenched to different levels by the different quinones:juglone (5-OH, 1,4 naphthoquinone), 1,4 naphthoquinone (1,4-NQ), and 1,2 naphthoquinone (1,2-NQ) considerably quenched the fluorescence of zeta-crystallin, where as the commonly used substrate, 9,10-phenanthrenequinone (PQ) did not induce significant quenching. Activity measurements showed only PQ served as a substrate for camel lens zeta-crystallin, while juglone, 1,4-NQ, and 1,2-NQ were inhibitors. Thus quinones that interacted with zeta-crystallin directly inhibited the enzyme, whereas the substrate had very low affinity for the enzyme in the absence of NADPH. Another substrate, dichlorophenol indophenol (DCIP), conformed to the same pattern; DCIP did not quench the fluorescence of the enzyme significantly, but served as a substrate. This pattern is consistent with an ordered mechanism of catalysis with quinone being the second substrate. All three naphthoquinones were uncompetitive inhibitors with respect to NADPH and noncompetitive with respect to PQ. These kinetics are similar to those exhibited by cysteine- and/or lysine-modifying agents. Juglone, 1,4-NQ, and 1,2-NQ interacted with and quenched the fluorescence of camel lens alpha-crystallin, but to lesser extent than that of zeta-crystallin.

Proteins that bind calcium in a phospholipid-dependent manner.

Three proteins (Mr = 64K, 32K, and 22K) that bind to phospholipids in a calcium-dependent manner were purified from bovine brain. The calcium-binding properties of these proteins were investigated by equilibrium dialysis and by gel filtration chromatography. The 64- and 32-kDa proteins were found to have calcium- and phospholipid-binding properties strikingly similar to those of protein kinase C [Bazzi, M.D., & Nelsestuen, G.L. (1990) Biochemistry 29, 7624]. The free proteins bound limited divalent metal ion even at 200 microM calcium. However, they bound eight to nine calcium ions per protein in the presence of membranes containing acidic phospholipids. The calcium concentrations needed for protein-phospholipid binding were different for these two proteins and were strongly influenced by the phospholipid composition of the vesicles; vesicles of higher phosphatidylserine content required lower concentrations of calcium for protein-membrane association. These properties described a general type of calcium-interacting system where simultaneous interaction of all three components (protein, phospholipids, and calcium) is required. The free proteins may provide only partial coordinate bonds to each calcium ion, but complete calcium-binding sites could be generated at the protein-phospholipid interface. In contrast to the 64- and 32-kDa proteins, the 22-kDa protein bound similar amounts of calcium (two to three ions/protein) in the presence or the absence of phospholipids. The 22-kDa protein had the lowest affinity for phospholipid and the highest affinity for calcium of the three proteins tested. Thus, calcium-dependent phospholipid-binding proteins consist of several types. For example, the 64- and 32-kDa proteins appear to be quite abundant and may even function as a calcium buffer to modulate signaling events.

Activation and regulation of protein kinase C enzymes.

Protein Kinase C (PKC) has been a principal regulatory enzyme whose function has been intensely investigated in the past decade. The primary features of this family of enzymes includes phosphorylation of serine and threonine residues located on basic proteins and peptide in a manner that is stimulated by calcium, phospholipid, and either diacylglycerol or phorbol esters. An additional intriguing feature of the enzymes is its ability to form two membrane-associated states, one of which is calcium dependent and reversible and the second is an irreversible complex which has the characteristics of an intrinsic membrane protein. Formation of the irreversible membrane-bound form is greatly facilitated by calcium and the tumor-promoting phorbol esters but does not appear to include covalent changes in the PKC structure. The intrinsic membrane-bound form is a very different enzyme in that its activity is no longer dependent on the other cofactors. It is proposed that formation of the irreversible membrane-bound form may be a mechanism for generating long-term cell regulation events where transient cell signals and second messengers induce long-term changes in the distribution of an enzyme in the cell. This property may be common to a number of regulatory proteins that are known to be distributed between the cytosol and membrane-fractions in the cell. Unfortunately, many problems have confronted study of PKC mechanism using the in vitro assay. This assay involves aggregation of the substrate, phospholipid, and enzyme to form a discontinuous mixture. Such a complex system prevents straightforward interpretation of enzyme kinetic data. Although many compounds affect the in vitro activity of PKC, most appear to accomplish this by relatively uninteresting mechanisms such as interference with the aggregation process. While some highly potent inhibitors undoubtedly interact directly with PKC, they also inhibit other enzymes and there are no entirely specific inhibitors of PKC known. Speculation on the possible roles of PKC in cell regulation are abundant and exciting. However, delineation of the regulatory roles of PKC may require another decade of intense effort.

Extensive segregation of acidic phospholipids in membranes induced by protein kinase C and related proteins.

Protein kinase C and two other proteins with molecular masses of 64 and 32 kDa, purified from bovine brain, constitute a type of protein that binds a large number of calcium ions in a phospholipid-dependent manner. This study suggested that these proteins also induced extensive clustering of acidic phospholipids in the membranes. Clustering of acidic phospholipids was detected by the self-quenching of a fluorescence probe that was attached to acidic phospholipids (phosphatidic acid or phosphatidylglycerol). Addition of these proteins to phospholipid vesicles containing 15% fluorescently labeled phosphatidic acid dispersed in neutral phosphatidylcholine resulted in extensive, rapid, and calcium-dependent quenching of the fluorescence signal. Fluorescence-quenching requirements coincided with protein-membrane binding characteristics. As expected, the addition of these proteins to phospholipid vesicles containing fluorescent phospholipids dispersed with large excess of acidic phospholipids produced only small fluorescence changes. In addition, association of these proteins with vesicles composed of 100% fluorescent phospholipids resulted in no fluorescence quenching. Protein binding to vesicles containing 5-50% fluorescent phospholipid showed different levels of fluorescence quenching that closely resemble the behavior expected for extensive segregation of the acidic phospholipids in the outer layer of the vesicles. Thus, the fluorescence quenching appeared to result from self-quenching of the fluorophores that become clustered upon protein-membrane binding. These results were consistent with protein-membrane binding that was maintained by calcium bridges between the proteins and acidic phospholipids in the membrane. Since each protein bound eight or more calcium ions in the presence of phospholipid, they may each induce clustering of a related number of acidic phospholipids.(ABSTRACT TRUNCATED AT 250 WORDS)