[Genetic dynamics of population systems represented by small subpopulations: analytical modeling and numeric results]. / Geneticheskaia dinamika populiatsionnykh sistem, predstavlennykh malymi subpopuliatsiiami: analiticheskoe modelirovanie i chislennye rezul'taty.

Genetic dynamics of population systems consisting of a finite number of small (Ne < 10(2)) semiisolated subpopulations was studied. A method of quantitative estimation of statistical parameters was developed for different types of population systems and different directions or intensities of selection. The following regularities were established: (1) optimal numbers of subpopulations, their effective size and rates of gene migration promoting continuous maintenance of genetic diversity can be chosen; (2) the genetic process in a population system is stationary only in the case of a specific structure of gene migrations corresponding to Wright's island model; (3) cyclic dynamics can stabilize the population system at high levels of gene diversity in a heterogeneous environment if gene migration and subpopulation size change in time. Similarities and differences between the concept of population system and the concept of metapopulation, which have been simultaneously proposed in Russia and abroad, are discussed in the final section.

Altered platelet-activating factor levels and acetylhydrolase activities are associated with increasing severity of bronchopulmonary dysplasia.

Lipid inflammatory mediators are thought to play an important role in the pathogenesis of neonatal lung injury and bronchopulmonary dysplasia (BPD). Because preliminary studies from the intensive care nursery of the University of Tennessee Medical Center, Knoxville, revealed linear increased in blood platelet-activating factor (PAF) levels in very low birthweight infants developing chronic lung disease and lower cord blood PAF acetylhydrolase activities in premature infants, it was theorized that altered platelet-activating factor levels and PAF acetylhydrolase activities are associated with increasing severity of BPD. Platelet-activating factor levels (blood and tracheal lavage) and PAF acetylhydrolase activities (blood and tracheal lavage) were measured over days 1 to 2, 3 to 5 and 6 to 7 in 16 ventilated infants and weekly in 9 infants with bronchopulmonary dysplasia. Platelet-activating factor values were normalized per nanogram of lavage blood urea nitrogen. Severity of bronchopulmonary dysplasia was estimated using the scoring system developed by Toce. Mean blood and lavage PAF levels and PAF acetylhydrolase activities were compared in infants developing bronchopulmonary dysplasia with those without the disease over the first seven days of life. Infants developing chronic lung disease were significantly smaller and of younger gestational age. In infants with bronchopulmonary dysplasia, higher PAF levels in blood were seen on days 3 to 5, along with increased lavage acetylhydrolase activities on days 1 to 2. Increased levels of PAF in lavage on days 3 to 5 were associated with increasing severity of bronchopulmonary dysplasia. Altered blood and lavage platelet-activating factor levels and PAF acetylhydrolase activities appear to be associated with the pathogenesis and severity of bronchopulmonary dysplasia.

Biosynthesis of platelet-activating factor and enzyme inhibitors.

Platelet-activating factor (PAF) is known to be synthesized by either a remodeling or de novo pathway. The enzymes responsible have been extensively studied by a number of laboratories. All evidence indicates the remodeling route is activated during inflammation and other hypersensitivity responses, whereas the de novo pathway is thought to be the source of PAF required for physiological functions. This article provides an update of what is currently known about the enzymatic systems that generate PAF as well as some preliminary findings we have obtained using potential inhibitors of the specific enzymes involved. Recent progress from our laboratory toward understanding the role of the CoA-independent and Co-A dependent transacylases in the formation of lyso-PAF and PAF is summarized.

Molecular species of sphingomyelin in sphingomyelinase-sensitive and sphingomyelinase-resistant pools of HL-60 cells.

This study of sphingomyelin molecular species in undifferentiated and differentiated (granulocytic form) HL-60 cells demonstrated only minor differences in the distribution of species between the sphingomyelinase-sensitive and sphingomyelinase-resistant pools of sphingomyelin in these cells. The two most prominent species of sphingosine present in both the undifferentiated and differentiated cells were those containing 16:0 (slightly higher in the sphingomyelinase-resistant membranes) and 24:1 N-acyl moieties. Cell differentiation exerted little effect on the distribution of molecular species of sphingomyelin between the sphingomyelinase-sensitive and sphingomyelinase-resistant pools in HL-60 cells, although the levels of N-palmitoyl sphinganine were significantly lower and the N-nervonoyl sphingosine higher in both pools from the differentiated cells. Our results indicate the same species of sphingomyelin, available at both the outer layer of the plasma membrane and inner layer of the plasma membrane (plus intracellular membranes) of HL-60 cells, serve as precursors for generation of the ceramides that participate in signal transduction processes initiated by cell activation.

Generation of the precursor (lyso-PAF) of platelet-activating factor via a CoA-dependent transacylase.

The microsomal fraction from rat spleen was shown to possess a CoA-dependent transacylase activity that produced 1-[3H]alkyl-2-lyso-sn-glycero-3-phosphocholine ([3H]lyso-PAF), the immediate precursor of PAF in the remodeling pathway of biosynthesis, from 1-[3H]alkyl-2-acyl-sn-glycero-3-phosphocholine. This CoA-dependent transacylase did not require ATP or metal ions for activity making it unlikely that either acyl-CoA-synthetase or a Ca(2+)-dependent phospholipase A2 were involved in the generation of [3H]lyso-PAF. Albumin, in addition to CoA, was required to demonstrate the formation of [3H]lyso-PAF from 1-[3H]alkyl-2-acyl-sn-glycero-3-phosphocholine. It appeared that a major function of albumin in the incubations was to complex the [3H]lyso-PAF formed, thus removing this end-product from the reaction.

The CoA-independent transacylase in PAF biosynthesis: tissue distribution and molecular species selectivity.

Microsomal membranes from six different rat tissues (spleen, lung, kidney, brain, testis, and liver) were found to possess CoA-independent transacylase activity that could both acylate lyso-[3H]PAF (1-[3H]hexadecyl-2-lyso-sn-glycero-3-phosphocholine) and then deacylate the 1-[3H]hexadecyl-2-acyl-sn-glycero-3-phosphocholine product via the transacylation of added exogenous 1-alk-1'-enyl-2-lyso-sn-glycero-3-phosphoethanolamine. Platelet-activating factor (1-[3H]hexadecyl-2-acetyl-sn-glycero-3-phosphocholine) was produced when acetyl-CoA was added to the spleen microsomes during generation of lyso-[3H]PAF by the transacylases. More extensive studies with subcellular fractions from spleen revealed that, in addition to microsomes, the transacylase activities were also present in the 15,000 x g membrane fraction but not in the cytosol. Analysis of molecular species of 1-[3H]hexadecyl-2-acyl-sn-glycero-3-phosphocholine before and after addition of 1-alk-1'-enyl-2-lyso-sn-glycero-3-phosphoethanolamine as the acyl acceptor demonstrated a high selectivity for polyunsaturated fatty acids (> 3 double bonds/acyl group) in both the acylation and deacylation processes that occurred in testicular microsomal membranes. The transfer of acyl groups by the transacylase appeared to be equally effective for either arachidonic or docosapentaenoic(n - 6) fatty acids, whereas linoleic and oleic fatty acids were not transferred from 1-[3H]hexadecyl-2-acyl-sn-glycero-3-phosphocholine following the addition of 1-alk-1'-enyl-2-lyso-sn-glycero-3-phosphoethanolamine. Similar experiments with the membrane fraction of undifferentiated HL-60 cells showed that arachidonic acid supplementation of intact cells enhanced both the CoA-independent transacylation of lyso-[3H]PAF and the subsequent deacylation of 1-[3H]hexadecyl-2-acyl-sn-glycero-3-phosphocholine caused by addition of 1-alk-1'-enyl-2-acyl-sn-glycero-3-phosphoethanolamine. Differentiation of the HL-60 cells into a neutrophil-like form had no effect on the transacylase activity. Our results indicate the PAF-related transacylase is widely distributed among tissues and, although highly selective for polyunsaturated acyl groups, does not discriminate selectively among the polyunsaturates.

Molecular species of ethanolamine plasmalogens and transacylase activity in rat tissues are altered by fish oil diets.

Effects of dietary fish oil ethyl esters and alkyldiacetylglycerols (an ether-linked lipid) on the distribution of subclasses of choline- and ethanolamine-glycerophospholipids as well as effects on highly unsaturated molecular species of ethanolamine plasmalogens from brain, spleen, kidney, lung, and testis of rats were examined. Supplementation of ethyl ester concentrates of n-3 fatty acids had no effect on the distribution of subclasses in any of the tissues. However, the supplements of 1-O-octadec-9'-enyl-2,3-diacetyl-sn-glycerol (diacetates of selachyl alcohol) caused significant increases in the alkylacylglycerophosphocholine and alkylacylglycerophosphoethanolamine subclasses from spleen and lung and in the alkylacylglycerophosphoethanolamine subclass from kidney. Dietary supplements of fish oil ethyl esters reduced the arachidonate-containing species of ethanolamine plasmalogens whereas molecular species having 20:5(n-3), 22:6(n-3), and/or 22:5(n-3) acyl groups were increased in the spleen, lung, and kidneys, but not brain. In testicular tissue from rats fed the fish oil diets, the molecular species of ethanolamine plasmalogens containing 22:5(n-6) acyl groups were reduced. An increase of ethanolamine plasmalogens with 18:1 alk-1-enyl moieties paired with highly unsaturated sn-2 acyl groups were found in the tissues of rats fed the fish oil plus selachyl alcohol diacetate supplements. Rats on the diet containing fish oil ethyl esters had significantly lower [3H]alkyllysoglycerophosphocholine CoA-independent transacylase activity in spleen microsomes than controls. This suggests that supplements of n-3 fatty acids interferes with the transacylation of arachidonate, an event that could seriously impair the release of arachidonate and lysophospholipids (e.g., lyso-PAF) that are precursors of potent bioactive lipid derivatives.

Reusable instruments are more cost-effective than disposable instruments for laparoscopic cholecystectomy.

Health care costs are rising rapidly, and surgeons can play a role in limiting costs of operations. Of the 600,000 cholecystectomies performed each year in the United States, approximately 80% are performed with laparoscopic technique. The purpose of this study was to compare the costs of reusable vs disposable instruments used during laparoscopic cholecystectomy. The costs to the hospital of reusable and disposable instruments were obtained. Instruments studied were the Veress needle, trocars and sleeves (two 10 mm and two 5 mm), reducers, clip appliers, and clips. In addition, the costs of sterilization and sharpening for reusable instruments were calculated. The cost of reusable instruments was based on an assumed instrument life of 100 cases. Data from three private hospitals and a Canadian university hospital were collected and examined. Data from the four hospitals revealed that the costs of reusable instruments per case were $46.92-$50.67. The comparable costs for disposable instruments were $330.00-$460.00 per case. Theoretical advantages of disposable instruments such as safety, sterility, and better efficiency are not borne out in literature review. In addition, the environmental impact of increased refuse from disposable instruments could not be exactly defined. With the consideration of significant cost savings and the absence of data demonstrating disadvantages of their use, reusable instruments for laparoscopic cholecystectomy, are strongly recommended.

Arachidonate-containing triacylglycerols: biosynthesis and a lipolytic mechanism for the release and transfer of arachidonate to phospholipids in HL-60 cells.

When HL-60 cells are incubated in media containing 10 microM [3H]arachidonic acid the label is immediately incorporated into both triacylglycerols and phospholipids. About one-half of the cellular tritium was associated with triacylglycerols after 2 h of incubation and this [3H]arachidonate was then transferred to phospholipids as soon as the labeled cells were placed in arachidonate-free media. A technique was devised to analyze the stereospecific distribution of [3H]arachidonate at the three sn-positions of glycerol in order to identify the mechanism(s) responsible for the biosynthesis of the labeled triacylglycerols. [3H]Arachidonate was found to be distributed in nearly equal amounts among all three glycerol positions of the triacylglycerols. In addition, analysis of intact triacylglycerols containing [3H]arachidonate revealed that 24% of the tritium eluted from reverse-phase HPLC with triarachidonoylglycerol. Both of these findings would be expected if a significant portion of the arachidonate-containing triacylglycerols were synthesized de novo. Homogenates prepared from [3H]arachidonate prelabeled HL-60 cells were capable of hydrolyzing the endogenous [3H]arachidonate-containing triacylglycerols to produce mainly free fatty acids and smaller amounts of monoacylglycerols. The relatively small amount of monoacyl- and diacylglycerols produced by the lipolytic activity of the homogenates indicated that [3H]arachidonate was hydrolyzed from all three sn-positions of the [3H]triacylglycerols. This lipase activity had a pH optimum of 4.5 and was associated to a greater extent with the soluble fraction than in the total membrane fraction. Although it is not known whether this lipolytic activity is the same as that expressed in the intact cells, the activity of the cell-free triacylglycerol lipase was of sufficient magnitude to have easily accounted for the decrease in [3H]triacylglycerols that was observed after transfer of the intact HL-60 cells (prelabeled with [3H]arachidonate) to fresh media. The data suggest that transfer of arachidonate from triacylglycerols to phospholipids probably occurs through an acyltransferase utilizing a lysophospholipid and arachidonoyl-CoA.

Endoscopic applications in thoracic surgery: video-assisted thoracoscopic surgery.

Twenty-eight cases of video-assisted thoracoscopic surgery were performed in a 10-month period. Twenty-seven of the 28 cases were completed without a standard thoracotomy. In one of the 28 cases, inadequate visualization secondary to extensive adhesions necessitated conversion to thoracotomy. There were no complications secondary to the video-assisted thoracotomy or video malfunction. Video-assisted thoracoscopic surgery provides the thoracic surgeon with advantages not enjoyed with standard thoracotomy and thoracoscopic techniques. These advantages include: decreased patient pain and recovery time, increased latitude of movement for the surgeon, visualization by assistant and observers, adjustable magnification for close examination of pathology and still photo and video recording capabilities.

Evidence for biosynthesis of plasmenylcholine from plasmenylethanolamine in HL-60 cells.

Both [3H]plasmenylethanolamine and [3H]plasmenylcholine were produced from substrates of [3H]alk-1-enylglycerol and [3H]alk-1-enyllysoglycerophosphoethanolamine by intact HL-60 cells. Molecular species analysis of the [3H]plasmenylcholine and [3H]plasmenylethanolamine formed indicated the major portion of plasmenylcholine originates from plasmenylethanolamine by a series of reactions catalyzed by phospholipase A2, lysophospholipase D, acyltransferase, phosphohydrolase, and cholinephosphotransferase. However, a significant but much smaller portion of the plasmenylcholine appeared to be synthesized from plasmenylethanolamine via a direct base-exchange or a coupled phospholipase C/cholinephosphotransferase reaction.

Reperfusion injury following testicular torsion and detorsion in prepubertal rats.

Acute testicular torsion is a surgical emergency which requires immediate intervention. Although damage to the gonad has been well documented, it remains unknown whether the majority of injury occurs during the period of torsion (ischemia) or following detorsion (reperfusion). The aims of this study were to determine: (1) whether damage following testicular torsion-detorsion has a reperfusion component similar to that described in other tissues, and (2) whether iron-catalyzed oxygen radical formation or altered calcium homeostasis plays a role in this injury. To study this, anesthetized prepubertal rats underwent 720 degrees intravaginal testicular torsion and were divided into groups of torsion only (ischemia) and torsion with reperfusion (ischemia/reperfusion). Reperfusion groups were treated prior to detorsion with either deferoxamine (iron chelator), diltiazem (calcium channel blocker), or saline vehicle. The results indicated that detorsion produces a qualitatively distinct reperfusion injury from that of non-reperfused testicles; however, such damage was not ameliorated by deferoxamine or diltiazem. Thus, testicular torsion-detorsion appears to have a significant reperfusion component that appears to not be mediated by iron-catalyzed oxygen radical formation or calcium injury.

Meats and fish consumed in the American diet contain substantial amounts of ether-linked phospholipids.

The primary goal of this study was to determine the amounts of ether-containing phospholipids, along with their concentration of certain polyunsaturated acyl groups, from selected, commonly consumed foods of animal origin (salmon, catfish, pork, beef, turkey and chicken). Levels of ether-linked glycerolipids in the samples were of particular interest, because ingestion of ether lipids could contribute to the production of platelet-activating factor (PAF; 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine), one of the most potent biological mediators known. Alkylacyl-sn-glycero-3-phosphocholine was found in all of the meats, with pork loin having the highest levels (0.9 mumol/g tissue) and chicken breasts the lowest (0.1 mumol/g tissue). Although choline plasmalogens were not as evident as the ubiquitous ethanolamine plasmalogens, substantial amounts (1.0 mumol/g tissue) of alk-1-enylacyl-sn-glycero-3-phosphocholine were found in tissues from beef and turkey. Triacylglycerols contained greater proportions of saturated fatty acids than phospholipids, and the ether-linked phospholipids were generally more unsaturated than diacyl species of the same phospholipid. Our data indicate that in addition to the phospholipid fraction of commonly eaten animal tissues supplying substantial amounts of polyunsaturated fatty acids, they are also a rich source of ether-linked lipids. Dietary ether-linked phospholipids could influence the lipid composition of host tissues to the extent that biological responses produced by ether lipid mediators would be affected.

Contributing factors in the trafficking of [3H]arachidonate between phospholipids.

Cultured human promyelocytic leukemia cells (HL-60), depleted of arachidonic acid by continued growth in serum-free media, were used as a model system to examine various factors that control the incorporation and distribution of [3H]arachidonic acid into classes and subclasses of cellular lipids. Increasing the culture media concentration of [3H]arachidonic acid from 1 x 10(-8) M to 1 x 10(-5) M caused a greater percentage of the cellular tritium to be distributed into triacylglycerols (from less than 1% at 1 x 10(-8) M to 38% at 1 x 10(-5) M) with a corresponding decrease in cellular [3H]diradylglycerophosphoethanolamine (from 53% at 1 x 10(-8) M to 12% at 1 x 10(-5) M) during 2 h incubations. A greater proportion of the tritium present in diradylglycerophosphoethanolamine and diradylglycerophosphocholine, at the higher media concentration of [3H]arachidonic acid (1 x 10(-5) M), was found in the diacyl subclasses of these two lipids than was observed at the lower concentrations (less than 1 x 10(-6) M) of [3H]arachidonic acid. Significant amounts of diarachidonoyl molecular species were found in the phosphatidylethanolamine (10%) and phosphatidylcholine (15%) of HL-60 cells that were labeled for 2 h with 1 x 10(-5) M [3H]arachidonic acid. This was the only molecular species of phosphatidylcholine to completely disappear when prelabeled cells were placed in arachidonate-free media for 22 h. Prelabeling-chase experiments with 1 x 10(-5) M [3H]arachidonic acid were consistent with movement of [3H]arachidonate from triacylglycerols into diradylglycerophosphatides and from diacylphospholipids into ether-linked phospholipids. Increasing the concentration of HL-60 cells in the incubations influenced the distribution of [3H]arachidonic acid in cellular lipid classes in a manner analogous to decreasing the concentration of [3H]arachidonic acid in the media. Increasing the endogenous level of cellular arachidonate in phospholipid classes with supplements of unlabeled arachidonic acid changed the subsequent lipid class distribution of a low concentration (1 x 10(-8) M) of [3H]arachidonic acid to resemble results obtained with a much higher mass level of [3H]arachidonate in arachidonate depleted cells. HL-60 cells differentiated into granulocytes by treatment with dimethyl sulfoxide incorporated less [3H]arachidonic acid but had a greater proportion associated with alkylacylglycerophosphocholine and alk-1-enylacylglycerophosphoethanolamine than undifferentiated HL-60 cells.

Chemical synthesis and physiological activity of sulfonium analogues of platelet activating factor.

Phosphatidylsulfocholine (PSC), the sulfonium analogue of phosphatidylcholine (PC), occurs naturally in some diatoms. The replacement of the [formula; see text] group by a [formula; see text] results in an increase in the polar head group size in PSC relative to that of PC, consistent with the observed increase in permeability of PSC bilayers towards urea. It was of interest to see whether replacement of the [formula; see text] group in platelet activating factor (PAF) by an [formula; see text] group leads to any change in platelet aggregation or other physiological activity. Synthesis of the sulfonium analogue of PAF was carried out by suitable modifications of known procedures. The PAF-sulfonium analogue was found to have almost the same platelet aggregating activity as PAF itself, in the concentration range 1-20 microM, but a much lower activity in the range 0.01-1 microM. The analogue had little or no effect on the platelet aggregation activity of PAF when added in the concentration range 0.01-1 microM and had about half the hypotensive activity of PAF towards hypertensive CDF male rats. The sulfonium analogue, however, was much more cytotoxic to HL-60 cells than PAF itself, in the concentration range 0-15 microM; replacement of the acetate group by a benzyl group increased the cytotoxicity to the level of that of the methoxy analogue of PAF. Thus, replacement of the [formula; see text] group by a [formula; see text] group in the polar head group region of PAF results in a relatively small change in its platelet aggregation activity and a decrease in its hypotensive activity, but greatly increases its antitumor activity.