Biomedical subjects

E Baldi

Publications and source records attributed to E Baldi.

At least 55 records · Page 3Linked to original sources

PubMed1997

2-Deoxy-D-galactose effects on passive avoidance memorization in the rat.

2-Deoxy-D-galactose (do-gal) hinders glycoprotein fucosylation. This compound was intracerebroventricularly administered to male adult Wistar rats in order to assess whether it could exert amnesic effects on a passive avoidance response (PAR) to be learned in the light-dark box apparatus. Three experiments were performed. In the first, do-gal was administered immediately after the acquisition trials at three dosages (2, 4, and 8 mumol). It was found that only the 4-mumol dosage was followed by PAR disruption. In the second, do-gal was administered at the adequate dosage (4 mumol) either 30 min before the acquisition trial or 30 min before retrieval testing. It was found that only the preretrieval administration was followed by PAR impairment. In the third, do-gal (4 mumol) was administered in postacquisition, at increasing postacquisition delays (0.25, 1.5, 4, and 6 h). It was found that there was PAR disruption only after do-gal administration at the shortest (0.25 h) delay. The results confirm that in the rat, glycoprotein fucosylation is involved in some of the phases of memory trace processing, and they are discussed in relation to other findings in the rat and the chick.

Animals↗

PubMed1997

Analysis of mnemonic processing by means of totally reversible neural inactivations.

The irreversible lesions technique precludes the analysis of the possibly critical role played by discrete brain sites in the several distinct stages of mnemonic processing (acquisition, consolidation, retrieval) during which these may be specifically but transiently active. On the contrary, the reversible functional inactivation techniques, by means of stereotaxic local microinjection of active compounds, make it possible to suppress the neuronal function of a discrete volume of nervous tissue, for a pre-determined time, with the assurance of complete functional recovery within a known duration. This technique makes it possible to block the neural activity of a chosen neural site at a given stage of memory processing without any interference with the function of the same structure either during earlier or later stages of the same process. Thus, the reversible ablation results may provide information not only on the qualitative topographical but also on the quantitative temporal dimension of learning and memory. The technique employed to cause totally reversible neural inactivation is detailed. The employment of several agents to obtain functional inactivation is discussed. Of these, perhaps the safest and most manageable is tetrodotoxin when a fairly long functional inactivation (e.g., 1 h) is desired. The effects of a reversible inactivation can be quite easily and accurately assessed by observing the severity of the amnesic disruption, if any, of a conditioned response. In order to do this as well as possible, it is advantageous to employ a very simple behavioral paradigm. The passive avoidance response in the light-dark box apparatus fulfills this requirement. Moreover, this paradigm, being one-trial, provides the necessary condition of a single well-defined temporal beginning. The present protocol has been successfully employed in learning and memory research, to assess when the functional integrity of a given neural structure is necessary in order that a conditioned response may be acquired, consolidated or retrieved. The employment of this protocol in relation to the intrinsic functional characteristics of a given subcortical neural site is discussed.

Animals↗

PubMed1996

Role of dorsal hippocampus in acquisition, consolidation and retrieval of rat's passive avoidance response: a tetrodotoxin functional inactivation study.

By means of local administration of tetrodotoxin (TTX) fully reversible functional inactivation of rat's dorsal hippocampus (DH) was obtained in order to define the role of this structure in the memorization of a conditioned passive avoidance response (PAR). In Experiment 1, on permanently cannulated animals, TTX (10 ng in 1.0 microliter saline) or saline (1.0 microliter) was injected uni- or bilaterally in the DH, respectively 1 h before PAR acquisition, immediately after PAR acquisition, and 1 h before PAR retrieval, always performed 48 h after the acquisition trial. It was shown that both pre-acquisition and pre-retrieval DH uni- or bilateral blockades were followed by significant PAR retention impairment, while in post-acquisition only the bilateral blockade determined PAR retention impairment. In Experiment 2, on three different groups of rats, TTX (10 ng in 1 microliter) saline) was bilaterally administered, under general ketamine anesthesia (100 mg/kg), into the DH at different post-acquisition delays (0.25, 1.5, 6 h). Retrieval testing, 48 h after treatment, showed that post-acquisition bilateral DH blockade caused PAR impairment only when performed 0.25 or 1.5 h after acquisition. The results indicate a well defined mnemonic role of DH during the acquisition, consolidation and retrieval of PAR engram. The experimental evidence is discussed in relation to other reports and to DH connectivity with the medial septal area and the amygdala.

Animals↗

PubMed1996

Human sperm activation during capacitation and acrosome reaction: role of calcium, protein phosphorylation and lipid remodelling pathways.

Two processes, namely capacitation and acrosome reaction, are of fundamental importance in the fertilization of oocyte by spermatozoon. Physiologically occurring in the female genital tract, capacitation is a complex process, which renders the sperm cell capable for specific interaction with the oocyte. During capacitation, modification of membrane characteristics, enzyme activity and motility property of spermatozoa render these cells responsive to stimuli that induce acrosome reaction prior to fertilization. Physiological acrosome reaction occurs upon interaction of the spermatozoon with the zona pellucida protein ZP3. This is followed by liberation of several acrosomal enzymes and other constituents that facilitate penetration of the zona and exposes molecules on the sperm equatorial segment that allows fusion of sperm membrane with the oolemma. The molecular mechanisms and the signal transduction pathways mediating the processes of capacitation and acrosome reaction are only partially defined, and appear to involve modifications of intracellular calcium and other ions, lipid transfer and phospholipid remodelling in sperm plasma membrane as well as changes in protein phosphorylation. The human and mouse sperm receptor for ZP3 has been recently sequenced and cloned. This receptor exhibits sequence homology with proto-oncogenes that mediate proliferation and differentiation in somatic cells. This review summarizes the main signal transduction pathways involved in capacitation and acrosome reaction.

Acrosome Reaction↗

PubMed1996

Platelet-activating factor enhances production of insulin-like growth factor binding proteins in a human adenocarcinoma cell line (HEC-1A)

We have recently demonstrated the existence of an autocrine growth loop driven by platelet-activating factor (PAF) in the human endometrial adenocarcinoma. cell line HEC-1A. To investigate a possible cooperation between PAF and the insulin-like growth factor (IGF) system in this cell line, the effect of PAF on insulin-like growth factor binding protein (IGFBP) production as well as binding and biological activities of IGF-I, IGFv-II, and the analog Des(1-3)IGF-I have been evaluated. Analysis of self- and cross-displacement curves of [125I]IGF-I binding to HEC-1A cells indicates the presence of a single class of binding sites, with affinity constants of 1-4 nM for IGF-I and IGF-II and 70 nM for Des(1- 3)IGF-I, which binds to IGFBPs with lower affinity. Insulin does not apparently bind to this binding site. Moreover, the addition of increasing concentrations of IGF-I leads to a paradoxical increase of binding. These results indicate a similarity of this binding site to IGFBPs. The presence of IGFBPs has been demonstrated by Western ligand blot analysis of HEC-1A conditioned medium which shows the presence of two bands of 32-34 and 40-45 kDa. By Western immunoblotting analysis, the two bands were respectively identified as IGFBP-2 and IGFBP-3. Incubation with PAF (1 microM) highly increases the release of the two IGFBPs from the cells. Such an effect is inhibited by the PAF receptor antagonist L659,989, by the PKC inhibitor sangivamycin, and by the tyrosine kinase inhibitor genistein. PAF also induces a time-dependent increase of mRNA expression for IGFBP-3, suggesting an effect on synthesis of this protein. IGF-I and IGF-II (0.1-100 nM) are almost ineffective in inducing [3H]thymidine incorporation, whereas a slight proliferative effect is observed with Des(1-3)IGF-I which also increases PAF synthesis. These data demonstrate a modulatory action of PAF on IGFBP secretion in HEC-1A cells and indicate that the IGF system plays a minor, if any, modulatory role on proliferation of this cell line.

Adenocarcinoma↗

PubMed1996

Amnesic effects of preacquisition, postacquisition, or preretrieval tetrodotoxin administration into the medial septal area on rat's passive avoidance memorization.

By means of local administration of tetrodotoxin (TTX) fully reversible functional inactivation of the medial septal area (MSA) of the rat was obtained in order to define the role of this structure in the retention of a conditioned passive avoidance response (PAR). In permanently cannulated animals, TTX (5 ng in 0.5 microl saline) or saline (0.5 microl) was injected in the MSA. TTX or saline was administered to six different groups of rats, respectively 1 h before PAR acquisition, immediately after PAR acquisition, and 1 h before PAR retrieval, performed 48 h after the acquisition trial. It was shown that MSA preacquisition and preretrieval TTX injections were followed by significant PAR retention impairment, while postacquisition TTX administration had no effect on PAR retention. The results indicate a well-defined mnemonic role of MSA during the acquisition and retrieval periods but not during the consolidation of PAR engram. The experimental evidence is discussed in relation to other reports and to MSA connectivity with other subcortical structures.

Animals↗

PubMed1996

Two functional assays of sperm responsiveness to progesterone and their predictive values in in-vitro fertilization.

We have recently reported, in a small cohort of subjects, that acrosome reaction (AR) and intracellular free calcium ([Ca2+]i) increase in response to progesterone were significantly correlated with in-vitro fertilization (IVF) rate. In the present study we extended these results to 90 subjects undergoing IVF. We confirm that both parameters were highly significantly correlated with the fertilization rate (P < 0.001). In particular, significantly lower responses to progesterone were detected in subjects with a fertilization rate < 50%, further enlightening the functional significance of sperm responsiveness to progesterone with respect to the process of fertilization. Moreover, we report here that both tests are highly discriminant of fertilization success, with positive predictive values > 90% for [Ca2+]i values which increase by > 1.2-fold and AR inducibility > 7% (cut-off values). Conversely, AR following challenge with the calcium ionophore A23187 was less significantly correlated with the percentage fertilization rate (P < 0.05), and showed lower predictive values than response to progesterone. All these tests ([Ca2+]i increase in response to progesterone, AR in response to progesterone and to A23187) appear highly sensitive and moderately specific. The positive predictive value may rise to > 95% when the combination of two tests ([Ca2+]i and inducibility of AR in response to progesterone) is considered. No correlation with fertilization rate has been found for spontaneous AR or basal [Ca2+]i. In conclusion, we propose that assessment of human sperm responsiveness to progesterone may be clinically useful in predicting fertilizing ability in vitro.

Acrosome↗

PubMed1996

Extracellular calcium negatively modulates tyrosine phosphorylation and tyrosine kinase activity during capacitation of human spermatozoa.

Capacitation of spermatozoa, a complex process occurring after sperm ejaculation, is required to obtain fertilization of the oocyte in vivo and in vitro. Although most of the biochemical/ biophysical events that occur during capacitation in vitro have been characterized, the molecular mechanisms underlying these complex events are still obscure. Increases of intracellular free Ca2+ concentrations ([Ca2+]i) and protein tyrosine phosphorylation have previously been demonstrated during in vitro capacitation of human spermatozoa. In the present study we investigated the relationship between extracellular/intracellular Ca2+, protein tyrosine phosphorylation, and tyrosine kinase and phosphatase activities during sperm capacitation. We report that the increase in tyrosine phosphorylation of several protein bands that occurs during sperm capacitation is independent of the presence of Ca2+ in the external medium and, at least partially, of the increase in [Ca2+]i occurring during the process. Indeed, the spontaneous increase in phosphorylation was still present in Ca(2+)-free/EGTA-containing-medium and in the presence of the intracellular Ca2+ chelator BAPTA/AM. Moreover, phosphorylation of proteins and protein tyrosine kinase (PTK) activity was enhanced if spermatozoa were incubated in Ca(2+)-free medium, suggesting the presence of Ca(2+)-inhibited tyrosine kinase(s) in human sperm. This hypothesis is further substantiated by the lower tyrosine phosphorylation observed after incubation with the ionophore A23187 and the endoplasmic Ca(2+)-ATPase inhibitor thapsigargin, which promote Ca2+ influx in human sperm. The ability of the cells to undergo acrosome reaction in response to progesterone, which can be considered a functional endpoint of capacitation, was highly compromised when spermatozoa were incubated in Ca(2+)-free medium or in the presence of EGTA, confirming that Ca2+ is required for sperm capacitation. Conversely, in the presence of erbstatin, a inhibitor of tyrosine kinase activity, which blunts tyrosine phosphorylation during capacitation, response to progesterone was maintained, suggesting that tyrosine phosphorylation must be kept at a low level (physiologically by the presence of Ca2+ in the external medium, or pharmacologically by the presence of erbstatin) in order to obtain response to progesterone. This mechanism may be important in vivo during sperm transit in the female genital tract to ensure appropriate timing of full capacitation in the proximity of the oocyte.

Acrosome↗

PubMed1996

Interferon-alpha downregulates expression of the oxytocin receptor in cultured human myometrial cells.

Previous studies in the endometrium of ruminants showed that type I interferon (IFN) prevents oxytocin receptor (OTR) formation. We studied the effect of IFN-alpha on human myometrial cells in culture expressing a high density of biologically active OTR. We found that IFN-alpha induced a 35-50% decrease in OTR mRNA and protein and that this inhibition was time and dose dependent. Maximal inhibition of OTR mRNA was obtained after 2-3 days, whereas 1-(beta-mercapto-beta, beta-cyclopentamethyl-enepropionic acid,2-O-Me-Tyr,Thr4,Orn8,Tyr9-amide)-[125I]vasotocin ([125I]OTA) binding reached a nadir after 3-4 days, with half-maximal inhibitory concentration (IC50) = 1,100 U/ml. Mathematical analysis of multiple homologous competition curves for [125I]OTA indicated that IFN-alpha treatment (5,000 U/ml x 3 days) reduced just the binding capacity (Bmax) without changing the binding affinity. Accordingly, the same treatment with IFN-alpha did not affect the half-maximally effective concentration (EC50) for the oxytocin-induced increase in intracellular calcium but significantly decreased maximal responsiveness (Emax) of myometrial cells to OT stimulation. In conclusion, our data demonstrate, for the first time, a negative regulation by IFN-alpha of the steady-state expression of OTR mRNA in cultured human myometrial cells obtained from nonpregnant uteri. This inhibition was followed by a parallel decrease in both the Bmax for [125I]OTA and Emax for oxytocin, suggesting a decreased OTR protein availability.

Base Sequence↗

PubMed1995

Tyrosine kinase inhibition reduces the plateau phase of the calcium increase in response to progesterone in human sperm.

Progesterone (P) has previously been shown to induce a rapid increase in [Ca2+]i as well as tyrosine phosphorylation of proteins in human spermatozoa. Both these effects are essential for induction of the acrosome reaction by P. We investigated a possible relationship between the P-induced calcium increase and tyrosine kinase activation, by evaluating the effect of the tyrosine kinase inhibitor genistein on these two effects. We found that preincubation with genistein abolished P-induced tyrosine phosphorylation of two sperm proteins of 97 and 75 kDa molecular weight and significantly inhibited the plateau phase of P-induced [Ca2+]i increase without affecting the peak phase. Conversely, the plateau phase was enhanced by the tyrosine phosphatase inhibitor Na3VO4. The effect of genistein was specific for P, since no inhibition was observed on the [Ca2+]i increase induced by thapsigargin, an inhibitor of endoplasmic Ca(2+)-ATPase previously shown to mobilize Ca2+ in spermatozoa. These results indicate that tyrosine kinase activation is involved in the generation of the plateau phase of Ca2+ influx induced by P, and suggest the possibility that two different pathways are involved in the induction of Ca2+ entry by P in human sperm.

Biological Transport↗

PubMed1995

Identification and characterization of functional angiotensin II receptors in human neuroblastoma cells.

The presence of specific AII receptors in 6 different human neuroblastoma cell lines was investigated using binding, cAMP and [Ca2+]i studies. We found high affinity (0.1 nM), low capacity ((1-2).10(3) sites/cell) binding sites for [125I](Sar-1,Ile-8)AII in one half of the cell lines studied. In the positive cell lines mathematical modeling of multiple competition curves among AII and analogs strongly indicated the presence of a homogenous class of sites, i.e., AT1 receptors. The presence of AT1 receptors was further substantiated by AII-induced inhibition of VIP-stimulated cAMP levels and by AII-evoked [Ca2+]i transient. The density of AT1 receptors in neuroblastoma cells was not affected by treatment with pertussis toxin and retinoic acid but was significantly increased by subacute treatment with VIP. In neuroblastoma cells, AII does not stimulate DNA synthesis, suggesting other roles rather than mitogenesis. Neuroblastoma cells represents an interesting model to investigate the function of AII in neural crest derived tissues.

1-Sarcosine-8-Isoleucine Angiotensin II↗

PubMed1995

Stimulation of protein tyrosine phosphorylation by platelet-activating factor and progesterone in human spermatozoa.

Tyrosine phosphorylation of proteins is involved in several sperm functions, including capacitation, motility, and acrosome reaction of spermatozoa. This study was undertaken to determine changes of tyrosine phosphorylation during 'in vitro' capacitation as well as the ability of platelet-activating factor (PAF) and progesterone (P), two known activators of sperm functions, to stimulate tyrosine phosphorylation of human sperm proteins. Spermatozoa were capacitated in BSA-containing medium and incubated with PAF (10-1000 nM) and progesterone (0.1-1 microgram/ml). After SDS-PAGE, sperm proteins were transferred to nitrocellulose and tyrosine phosphorylated proteins immunodetected by reacting with anti-phosphotyrosine antibody. The antibody mainly reacted with two proteins of approximately 97 and 75 kDa. The level of phosphorylation increased in these two proteins as a function of capacitation time, with a maximum between 120 and 180 min. In addition, phosphorylation in these two proteins was increased in capacitated spermatozoa by treatment with progesterone and PAF and was greatly reduced by pre-incubation with the tryosine kinase inhibitor erbstatin. Furthermore, pre-incubation with the two tyrosine kinase inhibitors erbstatin and genistein inhibited the induction of acrosome reaction by progesterone and, partially, by PAF. Our results suggest a role for tyrosine kinase(s) in the mechanism of capacitation and activation of human spermatozoa by PAF and progesterone.

Acrosome↗

PubMed1995

Time-dependent deficits of rat's memory consolidation induced by tetrodotoxin injections into the caudate-putamen, nucleus accumbens, and globus pallidus.

In order to better define corpus striatum involvement in memory processing, tetrodotoxin functional ablation was employed to assess the respective roles of the caudate-putamen as a whole and of its anterior, median, and posterior regions, of the nucleus accumbens, and of the globus pallidus during early passive avoidance response consolidation of the rat. Under general ketamine anesthesia (100 mg/kg) by means of microinjections, tetrodotoxin (10 ng/microliter) was bilaterally administered at the appropriate stereotaxic locations of the chosen sites and subsites at different postacquisition delays (0.25, 1.5, and 6 h). Saline solution was administered to control subjects in the same sites. Rats always underwent retrieval testing 48 h after treatment, i.e., when tetrodotoxin blockade had completely disappeared. The results show that both total caudate-putamen and median caudate-putamen blockades caused passive avoidance responding impairment when performed at 0.25- and 1.5-h postacquisition delays, that caudate-putamen anterior and caudate-putamen posterior blockades were never followed by passive avoidance impairment, and that both nucleus accumbens and globus pallidus blockades were followed by passive avoidance impairment when performed at 0.25- and 1.5-h postacquisition delays. In all structures, tetrodotoxin administration caused no impairment at the 6-h delay. The results are discussed with reference both to previous results obtained after reversible or irreversible lesions of the same structures and to the functional mnemonic relationships of the corpus striatum and its components with other subcortical structures, particularly the substantia nigra.

Animals↗

PubMed1995

Actions of progesterone on human sperm: a model of non-genomic effects of steroids.

Non-genomic actions of steroids have been extensively studied in the last few years. Among these actions, the non-genomic effect of progesterone (P) on human spermatozoa appears to be very promising, in view of the dramatic effect of this steroid on intracellular calcium, activation of tyrosine kinase, and induction of acrosome reaction. We have shown that the ability of spermatozoa to respond to P increases during the process of capacitation and is not counteracted by the P-receptor antagonist RU486 nor by the GABAA antagonists bicuculline and picrotoxin. We have also shown that P increases tyrosine phosphorylation of a sperm protein of about 97 kDa, suggesting activation of tyrosine kinase(s). In addition, we found that P induces a perturbation of sperm membrane phospholipid metabolism resulting in an increase of synthesis of platelet-activating factor and liberation of arachidonic acid. Results of these biochemical studies indicate that P is able to stimulate several signal transduction pathways in human sperm. We have also investigated responsiveness to P in sperm of oligozoospermic subjects as well as of men undergoing an in vitro fertilization (IVF) program. Our results show that the percentage increases of intracellular calcium and acrosome reaction in response to P is significantly reduced in oligozoospermic men as well as in subjects with reduced fertilization rate. Moreover, in the latter subjects response to P is highly significant correlated to fertilization rate of oocytes. These studies indicate that a biochemical alteration of sperm in their capacity to respond to P might be responsible for reduced fertilizing ability.

Acrosome↗

PubMed1995

Intracellular calcium increase and acrosome reaction in response to progesterone in human spermatozoa are correlated with in-vitro fertilization.

In this study we have investigated responsiveness to progesterone in spermatozoa from a group of unselected male partners of couples undergoing in-vitro fertilization (IVF). We evaluated progesterone-stimulated intracellular Ca2+ ([Ca2+]i) and percentage increase in acrosome reaction in the same sperm sample used for oocyte inseminations. [Ca2+]i was measured with a fluorimetric method, while the acrosome reaction was assessed using a fluorescent probe (fluorescein isothiocyanate-labelled peanut lectin). The average percentage [Ca2+]i as well as the rate of increase in the frequency of acrosome reaction following progesterone challenge were significantly lower (P < 0.005) in the group of patients with a fertilization rate < 50%. In addition, significant correlations between the fertilization rate and the progesterone-stimulated [Ca2+]i and acrosome reaction increases (r = 0.78 and r = 0.79 respectively) were observed. Furthermore, in cases of fertilization failure, no increase of [Ca2+]i or acrosome reaction was observed in response to progesterone with the exception of one case. Our results indicate that [Ca2+]i and acrosome reaction increases in response to progesterone can be of value in the prediction of sperm fertilizing ability. As the two parameters were significantly correlated to each other (r = 0.86), the two assays have similar IVF predictive value and might be used interchangeably as a diagnostic tool in the assignment of male patients to the different kinds of assisted fertilization techniques.

Acrosome↗

PubMed1995

Transitory effects of intracerebral administration of protein synthesis inhibitors on rat's spontaneous behavior.

In order to evaluate whether the effects of the local administration of protein synthesis inhibitors are transient or permanent, small volumes of cycloheximide, 2-deoxy-d-galactose, tetrodotoxin, and saline solution were stereotaxically administered in the amygdala, substantia nigra, and nucleus basalis magnocellularis of naive male adult Wistar rats. Three spontaneous activities (feeding, drinking, locomotion) were continuously recorded. 12 groups of animals were housed in activity cages for 6 consecutive days with a light/dark schedule (LD 12:12). The active compounds at doses sufficient to cause only limited effects were administered at the beginning of the light period of the fourth day. During the initial three days spontaneous activities remained constant, following the normal circadian distribution of rodents. Cycloheximide and tetrodotoxin administrations were followed by transient site-specific and activity-specific changes, which occurred only during the fourth and fifth day. The results show that protein synthesis inhibitors can be locally administered into the CNS at doses affecting only limited subcortical structures. The observed modifications of the three spontaneous activities were all transient and were always followed by complete recovery within 48 hours, thus confirming previous biochemical data. Therefore, the local injection of protein synthesis inhibitors, like tetrodotoxin, lidocaine or other compounds, can be employed to induce transient functional effects on well delimited brain structures in order to study the time-course of the mechanism of mnemonic trace formation.

Amygdala↗

PubMed1995

Energy expenditure (EE) and substrate utilization (SU) in the perioperative period in orthotopic liver transplantation (OLTX).

12 patients were investigated with IC (Datex, Deltatrac) preoperatively and during the surgical procedure of OLTX, VO2, VCO2 were continuously measured and RQ, EE and SU were calculated considering the different periods of the procedure: preoperative resting EE: (PREE), anaesthesiological procedures (ANEE), liver preparation EE (LPEEE), liver removal EE (LREE), anhepatic phase EE (APEE), reperfusion, EE (RPEE) and end of operation EE (EOEE). EE were expressed as % respect value calculated with H.B. (Harris-Benedict) formula. Data were analyzed with Student T-test and p < 0.01** or < 0.05* PREE is typical in end stage liver disease with low RQ values and increased EE. Energy production depends on lipid utilization since liver gluconeogenesis and glycogen stores are impaired. Anesthesia reduces energy needs and production up to 50% of the preoperative values, reducing VO2 more than VCO2 and therefore an augmented RQ value over 1.0. SU analysis indicates an increased glucose and aminoacid utilization coupled with high nitrogen catabolism that continues in the postoperative period (from 0.08 0.01 gN/kg b.w. to 0.20 0.06 gN/kg b.w.). When the new liver is reperfused, VO2 increases more than VCO2 indicating the risk of reperfusion injury.

Calorimetry, Indirect↗

PubMed1994

Platelet-activating factor mediates an autocrine proliferative loop in the endometrial adenocarcinoma cell line HEC-1A.

We investigated the synthesis and biological effects of platelet-activating factor (PAF) in the human endometrial cancer cell line HEC-1A. We found that HEC-1A cells actively synthesize and release PAF, as demonstrated by both [3H]acetate incorporation into PAF and gas chromatography-mass spectrometry studies. HEC-1A cells not only synthesize but also respond to PAF. Indeed, in fura-2-loaded cells, PAF stimulates [Ca2+]i increase with a median effective concentration of 5.6 nM. Furthermore, PAF induces a time-dependent expression increase of the nuclear protooncogene c-fos with a median effective concentration of 130 nM and stimulates DNA synthesis (median effective concentration, 700 nM). All of these effects are inhibited by the PAF receptor antagonist L659,989. Radioligand binding studies indicated the presence of two populations of PAF receptors with affinity constants in the nanomolar and micromolar range. Since the PAF antagonist per se inhibits DNA synthesis and cell proliferation, we suggest that PAF supports an autocrine growth circuit in HEC-1A cells. On the contrary, in the uterine leiomyosarcoma cell line SK-UT-1, which does not express specific binding sites for PAF, neither this phospholipid nor its receptor antagonist affect DNA synthesis. Our results provide evidence for the existence of an autocrine proliferative loop involving PAF in the endometrial cancer cell line HEC-1A.

Acetyl-CoA C-Acetyltransferase↗