Maternal avoidance, anxiety cognitions and interactive behaviour predicts infant development at 12 months in the context of anxiety disorders in the postpartum period.

BACKGROUND: Few studies have examined the relation between anxiety disorders in the postpartum period and cognitive as well as language development in infancy. AIMS: This longitudinal study investigated whether anxiety disorder in the postpartum period is linked to infant development at twelve months. A closer look was also taken at a possible link between maternal interaction and infant development. STUDY DESIGN: Subjects were videotaped during a Face-to-Face-Still-Face interaction with their infant (M = 4.0 months). Specific maternal anxiety symptoms were measured by self-report questionnaires (Anxiety Cognition Questionnaire (ACQ), Body Sensations Questionnaire (BSQ), Mobility Inventory (MI)) to check for a connection with infant development. The Bayley Scales of Infant and Toddler Development-III (Bayley-III) were used to assess infant language and cognitive development at one year of age. SUBJECTS: n = 34 mothers with anxiety disorder (SCID-I; DSM-IV) and n = 47 healthy mothers with their infant. OUTCOME MEASURES: Infant performance on Bayley-III language and cognitive scales. RESULTS: Infants of mothers with anxiety disorder yielded significantly lower language scores than infants of controls. No significant group differences were found regarding infant cognitive development. Exploratory analyses revealed the vital role of "maternal avoidance accompanied" in infant language and cognitive development. Maternal neutral engagement, which lacks positive affect and vocalisations, turned out as the strongest negative predictor of cognitive development. Maternal anxiety cognitions and joint activity in mother-infant interaction were the strongest predictors of infant language performance. CONCLUSIONS: Results underline the importance to also consider the interaction behaviour of women with anxiety disorders to prevent adverse infant development.

Maternal bonding in mothers with postpartum anxiety disorder: the crucial role of subclinical depressive symptoms and maternal avoidance behaviour.

Hardly any research has examined the link between postpartum anxiety disorder and maternal bonding. This study examined if postpartum anxiety disorder and maternal bonding are related in the postpartum period. Thereby, subclinical depressive symptoms and specific aspects of an anxious symptomatology were also taken into consideration. The German sample of N = 78 mother-infant dyads is composed of n = 30 mothers with postpartum anxiety disorders but without major or minor depression according to the Diagnostic and Statistical Manual of Mental Disorders IV (DSM-IV) and n = 48 healthy mothers. Subjects were interviewed with the Structured Clinical Interview for DSM-IV Disorders at an average infant age of M = 4.1 months. Moreover, mothers filled out the Postpartum Bonding Questionnaire-16. The Anxiety Cognitions Questionnaire, the Body Sensations Questionnaire and the Mobility Inventory were chosen to assess different aspects of anxious symptomatology. To control for concurrent subclinical depressive symptoms, we used the German Edinburgh-Postnatal-Depression Scale. Mothers with postpartum anxiety disorder reported significantly lower bonding than healthy mothers. However, in a linear regression analysis, concurrent subclinical depressive symptoms and avoidance of anxiety-related situations in company explained 27 % of the overall variance in maternal bonding. The perceived lower bonding of mothers with anxiety disorder could be due to aspects of a concurrent subclinical depressive symptomatology. This notion emphasizes the need to target even mild depressive symptoms in the treatment of postpartum anxiety disorders. The outcomes also underline that the severity of anxious symptomatology, reflected by avoidance behaviour in company, puts the mother-infant bond at risk.

Comparison of measured and simulated distribution of microbial biomass in subsurface vertical flow constructed wetlands.

The multi-component reactive transport module CW2D has been developed to model transport and reactions of the main constituents of municipal wastewater in subsurface flow constructed wetlands and is able to describe the biochemical elimination and transformation processes for organic matter, nitrogen and phosphorus. It has been shown that simulation results match the measured data when the flow model can be calibrated well. However, there is a need to develop experimental techniques for the measurement of CW2D model parameters to increase the quality of the simulation results. Over the last years methods to characterise the microbial biocoenosis in vertical subsurface flow constructed wetlands have been developed. The paper shows measured data for microbial biomass and their comparison with simulation results using different heterotrophic lysis rate constants.

Diversity of ammonia oxidising bacteria in a vertical flow constructed wetland.

Vertical flow constructed wetlands (VFCWs) with intermittent loading are very suitable for nitrification. Ammonia oxidising bacteria (AOB) are the limiting step of nitration. Therefore the AOB community of a full-scale VFCW, receiving municipal wastewater, was investigated within this study. The diversity of the functional gene encoding the alpha-subunit of the ammonia monooxygenase (amoA), present only in AOB, was assessed by denaturing gradient gel electrophoresis (DGGE). Only very few amoA sequence types dominated the wetland filter substrate; nevertheless a stable nitrification performance could be observed. During the cold season the nitrification was slightly reduced, but it has been shown that the same AOB could be identified. No spatial AOB pattern could be observed within the filter body of the VFCW. The most prominent bands were excised from DGGE gels and sequenced. Sequence analyses revealed two dominant AOB lineages: Nitrosomonas europaea/"Nitrosococcus mobilis" and Nitrosospira. Species of the Nitrosomonas lineage are commonly found in conventional wastewater treatment plants (WWTPs). In contrast, members of the Nitrosospira lineage are rarely present in WWTPs. Our observations indicate that the AOB community in this VFCW is similar to that found in horizontal flow constructed wetlands, but differs from common WWTPs regarding the presence of Nitrosospira.

[Magnetic resonance tomogography after cryotherapy of long bones in an animal model]. / Magnetresonanztomographie nach Kryoablation langer Röhrenknochen im Tiermodell.

PURPOSE: To describe the MR findings following cryoablation of long bones. MATERIALS AND METHOD: Cryoablation was performed in femoral and tibial bones of 24 sheep under general anesthesia. MRI of the treated and untreated contralateral bones was performed immediately thereafter and at 2, 4 and 6 months after the cryosurgical procedure. RESULTS: On the MRI performed immediately after cryotherapy, the lesions showed low signal intensities relative to the normal bone marrow on unenhanced T1- and T2-weighted images. At 2, 4 and 6 months after cryoablation, the lesions showed high signal intensities on STIR images, low signal intensities on T1-weighted and heterogeneous enhancement on contrast-enhanced T1-weighted MR images. The femoral lesions decreased in size from 31 +/- 3 mm immediately after the cryotherapy to 13 +/- 4 mm 6 month later and the tibial lesions from 29 +/- 7 mm to 19 +/- 4 mm. CONCLUSION: MRI shows bone marrow lesions immediately after cryotherapy and can easily monitor healing lesions. MR imaging is suitable for following cryotherapy.

JAK and STAT proteins are expressed and activated by IFN-gamma in rat pancreatic acinar cells.

The development of acute pancreatitis (AP) is triggered by acinar events, but the subsequent extra-acinar events, particularly a distinct immune response, appear to determine its severity. Cytokines modulate this immune response and are derived not only from immunocytes but also from pancreatic acinar cells. We studied whether pancreatic acinar cells were also capable of responding to cytokines. The JAK/STAT-pathway represents the main effector for many cytokines. Therefore, expression and regulation of JAK and STAT proteins were investigated in rat pancreatic acinar cells. Western blotting showed expression of JAK1, JAK2, Tyk2, and STAT1, STAT2, STAT3, STAT5, STAT6. In addition, STAT1 was reversibly tyrosine-phosphorylated upon the procedure of acinar cell isolation. In contrast, STAT3-phosphorylation occurred spontaneously after pancreas removal and was not reversible within 8 h. STAT1 phosphorylation was also observed upon treatment with IFN-gamma but not upon EGF, TNF-alpha or IL-6, and inhibited by the JAK2-inhibitor AG-490. Immunohistochemistry revealed cytoplasmic expression of unphosphorylated STAT1 in untreated acinar cells and nuclear translocation of phosphorylated STAT1 following IFN-gamma-treatment. Interestingly, although CCK leads to the activation of multiple stress pathways in pancreatic acinar cells, we found no influence of CCK on phosphorylation of STAT1, STAT3, or STAT5 in the pancreas. In conclusion, our data provide further evidence that pancreatic acinar cells are able to interact with immune cells. Besides stimulating immune cells via cytokine secretion, acinar cells are in turn capable of responding to IFN-gamma via JAK2 and STAT1 which may have an impact on the development of AP.

Response of Penaeus indicus females at two different stages of ovarian development to a lethal infection with Vibrio penaeicida.

An association between vitellogenesis and the immune system was suggested in crustaceans from studies on plasma lipoproteins. The present research studies the effect of an experimentally induced bacterial infection on vitellogenesis in females of the shrimp Penaeus indicus, as a model for penaeid species. Pre-vitellogenic and vitellogenic P. indicus females were experimentally infected with an extremely pathogenic bacterium, Vibrio penaeicida. The peak in mortality occurred earlier in pre-vitellogenic animals than in vitellogenic ones, although the final mortality level ( approximately 64-74%) 52h post-infection was nearly the same for the two groups. Twenty hours after infection, the total number of haemocytes was significantly reduced in vitellogenic females while there was no change in the pre-vitellogenic group. Protein synthesis in ovaries was not significantly affected by infection, at the two stages of ovarian development. No differences were found in mRNA levels of shrimp ovarian peritrophin protein (SOP), but preliminary results showed that mRNA expression of vitellin (VT) was reduced in a heavily infected vitellogenic female. The total amount of lipids in the haemolymph of vitellogenic females was almost twice higher than that of pre-vitellogenic ones. However, there was no change in the total content of lipids, lipid classes and fatty acid distribution in haemolymph or hepatopancreas following infection. Although vitellogenic and pre-vitellogenic females probably respond differently to a lethal bacterial infection, physiological differences may be concealed by the rapid onset of mortality.

Inhibition of de novo synthesis of a jelly layer precursor protein by crustacean hyperglycemic hormone family peptides and posttranscriptional regulation by sinus gland extracts in Penaeus semisulcatus ovaries.

Mature penaeid oocytes possess extracellular cortical rods (CR) that contain precursor proteins of the jelly layer (JL) that forms a protective layer around eggs immediately after spawning and dissipates following the assembly of the hatching envelope. The temporal pattern of protein synthesis and mRNA expression of a jelly layer precursor protein in Penaeus semisulcatus ovaries was followed during vitellogenesis, and the regulation by sinus gland extracts (SGE) and crustacean hyperglycemic hormone (CHH) family peptides was evaluated. An approximately 33-kDa jelly layer precursor protein was previously identified in ovaries, CR, and JL and was named shrimp ovarian peritrophin-like protein (SOP), because its deduced amino acid sequence shows structural similarities to insect peritrophins. SOP was synthesized in ovarian explant fragments that were removed from vitellogenic ovaries and incubated in vitro, but synthesis was not detected in explants that were collected from previtellogenic ovaries. SOP transcripts were detected in all stages of ovarian development, but were more abundant in previtellogenic ovaries than in other stages. De novo synthesis of SOP was inhibited by P. semisulcatus SGE and by CHH family peptides that were purified from P. japonicus sinus glands. Sinus gland extracts, however, did not affect the steady state levels of SOP transcripts at any stage of ovarian development. These results suggest that SGE regulate SOP synthesis at the posttranscriptional level.

Molecular characterization and high expression during oocyte development of a shrimp ovarian cortical rod protein homologous to insect intestinal peritrophins.

Penaeoid shrimp oocytes nearing the completion of oogenesis are enveloped in an acellular vitelline envelope and possess extracellular cortical rods (CRs) that extended into the cortical cytoplasm. These cortical specializations are precursors of the jelly layer (JL) of the egg. In searching for highly expressed mRNAs during oogenesis in the marine shrimp (Penaeus semisulcatus), two related cDNAs have been isolated that encode a mature protein of 250 amino acid residues. The deduced amino acid sequences revealed the presence of repeated cysteine-rich domains that are related to the chitin-binding domains of insect intestinal peritrophins. Similar cysteine-rich domains were reported in insect intestinal mucin, crustacean tachycitin, and invertebrate chitinases. The shrimp ovarian peritrophin (SOP) is glycosylated and can bind chitin when extracted from CRs. Its apparent molecular mass in SDS-PAGE is 29-35 kDa and 33-36 kDa, under nonreducing or reducing conditions, respectively. SOP is a major protein of CRs and the JL, and was immunodetected in ovaries; purified CRs; fertilized eggs that were surrounded by a JL matrix; and in the cloudy, whitish flocculent material appearing in sea water immediately after spawning. Immunolocalization in tissue sections determined that SOP was present in oocyte cytoplasm and in extraoocytic CRs. Shrimp expressed SOP mRNA in ovaries at all oocyte developmental stages, whereas expression in the hepatopancreas was restricted to vitellogenic stages. SOP mRNA was abundant in the shrimp ovary and was detected before the presence of the corresponding protein. This is the first demonstration that a protein with similar features to insect intestinal peritrophins is a component of CRs and is therefore a main precursor of the JL of spawned shrimp eggs.

The effects of short term lipid infusion on plasma and hepatic bile lipids in humans.

BACKGROUND: Patients on parenteral nutrition have an increased incidence of gall bladder sludge and gallstone disease, thought to be related to bile stasis. Intravenous lipid emulsions, especially those containing medium chain triglycerides, have also been shown to have a lithogenic effect on the composition of bile in the gall bladder. AIMS: To determine whether lipid infusion influences hepatic bile composition in patients with an indwelling T tube following cholecystectomy and choledochotomy. METHODS: In eight patients undergoing the above surgical procedure, the time at which effects of the interrupted enterohepatic circulation were minimal was determined. Twenty two cholesterol gallstone patients with bile fistula were then randomised to receive an infusion of a lipid emulsion containing either long chain triglycerides or a mixture of long and medium chain triglycerides. RESULTS: Lipid infusion resulted in a significant increase in plasma levels of triglycerides and phospholipids. Both lipid emulsions caused an increase in hepatic biliary cholesterol level and cholesterol saturation index, but this effect was more pronounced with medium chain triglycerides. The fatty acid composition of biliary phospholipids showed a significant enrichment of linoleic acid by both lipid infusions. CONCLUSIONS: Infusion of triglycerides causes lithogenic changes in hepatic bile composition in humans, the lithogenic effect of infusion of medium chain triglycerides being more pronounced than that of long chain triglycerides. This effect, coupled with gall bladder stasis, may be responsible for the increased risk of biliary sludge and gallstone formation in patients on long term lipid infusion.

The incorporation of fatty acids of different chain length into liver and biliary lipids in the perfused rat liver.

In an attempt to correlate the incorporation of fatty acids (FA) of different chain length into liver and biliary lipids, isolated rat livers were perfused for 2 h with Krebs-Ringer bicarbonate containing 1% albumin and 10 mumol of [1-14C]-labeled FA: C2, C8, C10, C12, C16, and C18:1. One to 1.36 mumol of medium-chain fatty acids (MCFA, C8, C10, and C12) and 6.6 mumol of long-chain FA (LCFA) were incorporated into liver lipids, 40% of the latter into phosphatidylcholine (PC). 14C-acetate (13 nmol) was incorporated into biliary cholesterol; 14C-MCFA contributed only 3.2-5 nmol; LCFA did not lead to newly synthesized cholesterol. Newly synthesized liver PC (2.75 to 3.25%) and newly synthesized liver cholesterol (6.5 to 10%) were secreted into bile. The specific radioactivity of biliary PC after infusion of all-saturated FA was 3.8-6.8 times higher than that of liver PC; for C18:1 it was only 1.7-fold. The specific radioactivity of biliary cholesterol, as compared to liver cholesterol, was 12 times higher for C2 and five times higher for MCFA. This indicates that a considerable proportion of the newly synthesized lipids was secreted into bile prior to significant mixing with preexisting liver PC and cholesterol pools. Liver PC contained 8% of unchanged 14C-C12; while 14C-C10 was not detected. Biliary PC, in contrast, contained 18% of unchanged 14C-C12 and 3% 14C-C10. These results suggest that after prolonged infusion of medium-chain triacylglycerols/long-chain triacylglycerols to patients, biliary PC may become enriched with MCTA. In addition, the oxidation of these FA may provide C-2 units which increase cholesterol synthesis.

Lipid accumulation in the ovaries of a marine shrimp penaeus semisulcatus (de haan)

By the end of oocyte development, the ovaries of Penaeus semisulcatus have accumulated almost equal amounts (approximately 16 mg lipid g-1 protein) of phospholipids and triacylglycerols. The phospholipids consist mainly of phosphatidylcholine (75-80 %) and phosphatidylethanolamine (20-25 %). Approximately 30 % of the total fatty acid content of both phospholipids and triacylglycerols is made up of polyunsaturated fatty acids. In fractions obtained by centrifugation of ovarian homogenates, most of the increase in levels of ovarian lipids during ovarian maturation was associated with an increase in triacylglycerol levels in the floating fat fraction and of phospholipids in the infranatant fraction. The presence of polyunsaturated fatty acids in the ovaries indicates the occurrence of lipid transport to the ovary during oocyte maturation. The gradual decrease in the relative abundance of polyunsaturated fatty acids as the ovaries matured supports previously published results suggesting intra-ovarian synthesis of saturated and mono-unsaturated fatty acids. Most of the lipids found in the female haemolymph (64.8 %) were recovered in the high-density lipoprotein fraction after density ultracentrifugation. The haemocyanin fraction recovered from this stage of fractionation contained substantial amounts of lipid (16.8 %) that could be removed by further sequential centrifugation at a higher NaBr density, leaving less than 0.9 % of the total haemolymph lipids associated with this fraction. While 16.2 % of the lipids were recovered from the very high-density lipoprotein fractions, these lipoproteins carried only 64-89 microg lipid mg-1 protein compared with 538.9 microg lipid mg-1 protein in the high-density lipoprotein fraction, indicating that the high-density lipoproteins are more likely to be the main transporters of lipids to the ovary. However, the contribution of very high-density lipoproteins to lipid transport cannot be ruled out at this stage. In this study, we present two models for lipid transport to the ovary based on the abundance of phospholipids and triacylglycerols in the haemolymph and on the amounts of polyunsaturated fatty acids accumulated within the ovary during vitellogenesis.

Hyperglycaemic hormones inhibit protein and mRNA synthesis in in vitro-incubated ovarian fragments of the marine shrimp Penaeus semisulcatus.

The present work shows for the first time that peptides belonging to the Crustacean hyperglycaemic hormone family (CHH-family hormones) from Penaeus japonicus affect protein and mRNA synthesis in in vitro-incubated ovarian explant fragments removed from vitellogenic females of Penaeus semisulcatus. Reduced levels of protein synthesis, determined by TCA-precipitable 35S-labeled proteins, were found in the presence of crude sinus gland extracts from both P. semisulcatus and P. japonicus. A similar inhibitory effect compared to controls was found with each of the seven CHH-family peptides. Non-CHH-family peptides did not reduce protein synthesis. Crude sinus gland extracts prepared from P. semisulcatus were at least 20-fold more effective than sinus gland extracts of P. japonicus. The inhibition level was directly related to the concentration of the peptide in the incubation media, but its degree varied among the different tested peptides. The profile of proteins synthesized during in vitro incubation was analyzed using polyacrylamide gel electrophoresis under denatured and reduced conditions (SDS-PAGE), followed by autoradiography. Synthesis of several proteins was reduced, including proteins with electrophoretic mobility similar to that of vitellin. Immunoprecipitation with antiserum prepared against native ovarian vitellin confirmed the inhibitory effect of CHH-family peptides on vitellin synthesis. The crude sinus gland extract and CHH-family peptides also inhibited RNA synthesis, as determined by [3H]uridine incorporation into mRNA of ovarian fragments. It is concluded that in addition to their role in carbohydrate metabolism, CHH-family peptides may also influence ovarian physiology in crustaceans.

Diagnosis of pulmonary Kaposi's sarcoma by detection of human herpes virus 8 in bronchoalveolar lavage.

Human herpes virus 8 (HHV8) DNA has recently been detected in sarcoma tissue of patients with Kaposi's sarcoma. HHV8 DNA could also be found in bronchoalveolar lavage (BAL) fluid of patients with tracheobronchial Kaposi's sarcoma. To determine the specificity, sensitivity and predictive values of HHV8 DNA detection in the BAL for the diagnosis of pulmonary Kaposi's sarcoma, 100 consecutive BAL were prospectively analyzed for the presence of HHV8 DNA using a nested PCR assay. In addition, 19 BAL samples of 14 AIDS patients with cutaneous or visceral Kaposi's sarcoma were retrospectively investigated. The prospective group consisted of 79 BAL performed in immunocompromised and of 21 BAL in nonimmunocompromised patients. Four patients of the prospectively analyzed group undergoing six BAL showed tracheobronchial Kaposi's sarcoma at five bronchoscopies. All of the five BAL samples performed in these patients with endoscopically visible Kaposi's sarcoma were positive for HHV8 DNA. Following chemotherapy and antiretroviral treatment tracheobronchial Kaposi's sarcoma was no longer detectable at a subsequent bronchoscopy and HHV8 DNA in BAL became negative in one patient. One BAL sample of a HIV-positive patient with no evidence of Kaposi's sarcoma was HHV8 DNA-positive. The sensitivity, specificity, positive and negative predictive values of HHV8 detection for the diagnosis of tracheobronchial Kaposi's sarcoma were 100%, 98.9%, 83.3%, and 100%, respectively. Twelve of 19 BAL samples of the retrospective group were HHV8 DNA-positive. In this group, 10 patients undergoing a total of 14 BAL suffered from pulmonary Kaposi's sarcoma. HHV8 DNA was documented in 10 of these 14 BAL samples. In three BAL of this group HHV8 DNA was positive, but pulmonary Kaposi's sarcoma was diagnosed at a later stage. In conclusion, the detection of HHV8 DNA in BAL is restricted to patients with Kaposi's sarcoma and is highly sensitive and specific for pulmonary involvement of Kaposi's sarcoma.

Eosinophilia in patients infected with the human immunodeficiency virus.

The prevalence and significance of peripheral blood eosinophilia in patients infected with the human immunodeficiency virus (HIV) were evaluated. Fifteen of 119 consecutive patients had absolute eosinophil counts of > 450/mm3. During a mean follow-up period of 419 days eosinophilia could be identified as secondary to a parasitic infection in only one patient. Correlation with disease stage showed a higher rate of advanced disease in patients with absolute eosinophilia. In a multivariate regression analysis, only low CD4+ cell counts, not the CDC disease stage or the use of antiretroviral therapy or primary prophylaxis, contributed significantly to the prevalence of eosinophilia. It is concluded that expensive laboratory investigations in asymptomatic patients with advanced-stage HIV disease are neither necessary nor cost effective.

A study of desmopressin and blood loss during spinal fusion for neuromuscular scoliosis: a randomized, controlled, double-blinded study.

BACKGROUND: Studies examining the use of desmopressin acetate (DDAVP) have shown variable results in DDAVP's efficacy for reducing blood loss. Studies of adults having cardiac surgery and of children having spinal fusion have suggested that patients with complicated medical histories and complex surgical procedures may benefit from use of DDAVP. Therefore, this study was designed to examine the homeostatic effects of DDAVP in children with severe cerebral palsy undergoing spinal fusion. METHODS: A randomized, double-blinded, and placebo-controlled trial of DDAVP was designed to enroll 40 patients. However, termination of the study was advised by the Institutional Review Board after 21 patients were enrolled. All patients had spastic quadriplegic-type cerebral palsy and were randomly assigned to one of two groups. The DDAVP group received 0.3 microg/kg DDAVP in 100 ml normal saline, and the placebo group received normal saline alone. All patients were anesthetized with nitrous oxide, oxygen, isoflurane, and fentanyl. Factor VIIIC and von Willebrand's factor (vWF) concentrations were measured in blood drawn before DDAVP infusion and 1 h after infusion. Blood pressure was maintained at a systolic pressure of less than 100 mmHg. Use of crystalloids, packed erythrocytes, platelets, and fresh frozen plasma were based on criteria established by protocol. Estimated blood loss was assessed by weighing sponges and measuring suctioned blood from canisters. RESULTS: Estimated blood loss (intraoperative and postoperative) and amount of packed erythrocytes transfused were similar for the DDAVP and placebo groups. Concentrations of both factor VIIIC and vWF were significantly greater after DDAVP infusion when compared with concentrations after placebo infusion. CONCLUSIONS: In the children who had complex spinal fusion, there was no difference in estimated blood loss between those who received DDAVP and those who received a placebo. Administration of DDAVP significantly increased factor VIIIC and vWF levels.

Isolation and characterization of the high-density lipoproteins from the hemolymph and ovary of the penaeid shrimp Penaeus semisulcatus (de Haan): apoproteins and lipids.

The high-density lipoproteins (HDLs) found in the male and female hemolymph of Penaeus semisulcatus de Haan were isolated by NaBr (1.22 g/ml) followed by sucrose gradient (5-25%) ultracentrifugation. The male HDL contained one protein, lipoprotein 1 (LP1), composed of one 110-kDa peptide subunit. The female HDL contained two proteins: 1) the LP1 that was immunoidentical to the male LP1 and was similarly composed of one 110-kDa peptide subunit and 2) vitellogenin (Vg), reacting positively with the rabbit antiserum generated against vitellin (Vt) that was isolated from vitellogenic ovaries. Both Vg and Vt consisted mainly of three polypeptide subunits (200, 120, and 80 kDa) as revealed by denatured PAGE and Western blot. The LP1 from males or females did not react with the Vt rabbit antiserum. Similarly, Vg and Vt did not react with the rabbit antiserum prepared against LP1. Phospholipids (PL) constituted 71-76% of the total lipids in the hemolymph and HDLs of both male and female hemolymph. Cholesterol (Ch) amounted to 17-20%, and small amounts (5%) of diacylglycerols (DAG) were also carried by these HDLs. Both the PL and DAG contained highly unsaturated fatty acids (20:5 omega 3 and 22:6 omega 3) that are transported from the food or hepatopancreas to the tissues, including the vitellogenic ovaries in females. In the present study we show for the first time the separate lipid composition of female LP1 and Vg and compare them with the lipids attached to the Vt. Vg had a lower lipid content than LP1 (540 and 1089 mg/g protein, respectively). Differences were also found in the relative abundance of PL, Ch, and DAG classes in the LP1 in comparison with Vg. Furthermore, small amounts (approximately 3.8%) of triacylglycerols (TAG) were found only in the hemolymph of vitellogenic females, and they were associated with the Vg. Although Vg and Vt were composed of similar polypeptides, their lipid composition was different Vt, in contrast to Vg, carried considerable amounts of TAG (approximately 22%) and only trace amounts of DAG. The significance of the TAG in the hemolymph of vitellogenic females is not known, and the functional relationship between Vg and Vt requires future extensive studies. Lipids were not detected in hemocyanin that was purified from clotted hemolymph.

Biosynthesis of medium-chain triacylglycerols and phospholipids by HepG-2 cells.

In an attempt to understand the metabolism by the liver of fatty acids (FA) of different chain length, we have studied the incorporation of [1(-14)C]-labeled C2, C8, C10, C12, and C16 into cellular lipids by HepG-2 cells. Over 90% of the radiolabeled FA were detected in phospholipids (PL) and triacylglycerols (TAG). The incorporation of C12 and C16 was three to four times higher than that of C8 and C10 (and reached 35 nmoles per mg protein after 1.5 h). The radioactivity of C2, C8, and C10 was recovered mainly in PL. C12 and C16 were incorporated at approximately equal amounts into PL and TAG. The radioactivity of both C2 and C8 was recovered exclusively in long-chain FA, suggesting oxidation of C8 into C2 units prior to FA synthesis. C10 likewise yielded mainly long-chain FA. However 10% of unchanged C10 was found in PL and up to 30% in TAG. 14C-C12 was largely incorporated unchanged. Under these conditions, the presence of C10 and C12 in PL and TAG was shown also by gas-liquid chromatography. In the presence of either C2, C8, or C10, up to 30% of 14C-monounsaturated FA were detected in PL and TAG. With C12 and C16, the fraction of 14C-monounsaturated FA was much smaller suggesting that extensive desaturation occurred during de novo synthesis.