Biomedical subjects

Z Cai

Publications and source records attributed to Z Cai.

At least 55 records · Page 3Linked to original sources

PubMed2001

Effect of heat treatment on mechanical properties of age-hardenable gold alloy at intraoral temperature.

OBJECTIVE: The aim of this study was to investigate the effect of various heat treatments on the mechanical properties of gold alloys capable of age-hardening at intraoral temperature. METHODS: Dumbbell-shaped patterns (ISO 6871) were cast with three gold alloys (Sofard; NC Type-IV; Aurum Cast, NihombashiTokuriki Co.). The Sofard alloy is age-hardenable at intraoral temperature. The castings underwent various heat treatments [as-cast (AC); solution treatment (ST); high-temperature aging (HA); intraoral aging (IA)]. After these heat treatments, ultimate tensile strength (UTS), 0.2% offset yield strength (YS), and elongation (EL) were measured at a strain rate of 1.7x10(-4)/s. Fracture surfaces of the specimens after tensile testing were observed using SEM. Vickers hardness was also measured after heat treating. RESULTS: After IA, the hardness values of the Sofard alloy increased and reached values similar to the hardness of the Sofard specimens aged at high temperature (HA). The hardness values of the NC Type-IV and Aurum Cast specimens slightly increased after IA, but did not reach the values of the specimens after HA. All the Sofard, NC Type-IV and Aurum Cast specimens showed significantly (P<0.05) greater hardness values after HA, compared with the values after any other heat treatments (AC, ST and IA). The UTS and YS of the specimens indicated a tendency similar to the results obtained for hardness. The Sofard specimens with ST showed the greatest elongation compared to the corresponding NC Type-IV and Aurum Cast specimens. However, the elongation of the Sofard specimens was abruptly reduced after intraoral aging. CONCLUSION: Intraoral aging significantly improved the mechanical properties and hardness of the Sofard alloy.

Analysis of Variance↗

PubMed2001

Porcelain adherence to dental cast CP titanium: effects of surface modifications.

OBJECTIVES: A reaction layer forms on cast titanium surfaces due to the reaction of the molten titanium with the investment material. Such a layer prevents strong adhesion between titanium and porcelain. This study characterized the effects of surface modifications on cast titanium surfaces and titanium-ceramic adhesion. METHODS: ASTM grade II CP titanium was cast into an MgO-based mold. Castings were devested by sandblasting with alumina particles, and subjected to surface modification by immersion in one of the following solutions: (1) 35% HNO3-5% HF at room temperature for 1min; (2) 50% NaOH-10% CuSO4 x 5H2O at 105 degreesC for 10min; (3) the NaOH-CuSO4 solution followed by the HNO3-HF solution; (4) 50% NaOH-10% NaSO4 at 105 degreesC for 10 min; (5) the NaOH-NaSO4 solution followed by the HNO3-HF solution; and (6) 50% NaOH solution at 105 degreesC for 10min. Surfaces only sandblasted with alumina were used as controls. Specimen surfaces were characterized by XRD and SEM/EDS, and hardness-depth profiles were determined. All specimens were sandblasted with 110 microm alumina particles before porcelain firing. An ultra-low-fusing porcelain (Vita Titankeramik) was fused on the titanium surfaces. The titanium-ceramic adhesion was characterized by a biaxial flexure test, and area fraction of adherent porcelain (AFAP) was determined by X-ray spectroscopy. RESULTS: EDS analyses revealed a substantial amount (13-17 wt%) of Al on the control, and specimens modified with Methods 2, 4, and 6. XRD revealed residual stress in the titanium surfaces and corundum on the control, and Methods 2, 4, and 6 specimens. A new Ti(Cu, Al)2 phase was identified on the titanium surfaces modified by immersion in 50% NaOH-10% CuSO4 x 5H2O aqueous solution. Reduced residual stress was observed on Method 1, 3, and 5 specimens. No corundum peaks were detected on these specimens. Compared to the control, significantly lower (P < 0.05) hardness values were found for Methods 1-3 and Method 5 specimens at 20 microm below the surfaces and for Method 1-5 specimens at 50 microm below the surfaces. Significantly higher (P < 0.05) AFAP values were found for surfaces modified with Methods 2-6 compared to the control and Method 1, and no significant differences were found among Methods 1-6, and between control and Method 1. SIGNIFICANCE: Based on the results from the present study, porcelain adherence to cast CP titanium can be improved by the caustic baths used in the study.

Dental Casting Investment↗

PubMed2001

X-ray diffraction characterization of dental gold alloy-ceramic interfaces.

X-ray diffraction (XRD) was employed to study dental alloy-ceramic interfaces. A Au-Pd-In alloy, which requires oxidation before porcelain firing, and a Au-Pt-Pd-In alloy, which does not require oxidation before porcelain firing, were selected in this study. Alloy specimens were centrifugally cast. Specimen surfaces were metallographically polished through 0.05 microm Al2O3 slurries. A thin layer (< 50 microm) of a dental opaque porcelain was fired on the alloy surfaces with and without initial oxidation. XRD was conducted at room temperature on four types of alloy specimens: polished, oxidized, porcelain fired after alloy oxidation, and porcelain fired without initial alloy oxidation. XRD was also performed on fired opaque porcelain without an alloy substrate. The detection of prominent gold solid solution peaks from alloy-ceramic specimens indicated that the incident X-ray beam reached the alloy-ceramic interface. In2O3 and beta-Ga2O3 were identified on the oxidized Au-Pd-In alloy, while In2O3 and SnO2 were detected on the oxidized Au-Pt-Pd-In alloy. Preferred orientation was observed for all the oxides formed on the alloys. Minimum lattice parameter changes (<1%) for the gold solid solutions were observed for both alloys before and after oxidation and porcelain firing. Leucite (KAlSi2O6, TiO2, ZrO2 and SnO2 were detected on the fired opaque porcelain. For both alloys, no additional oxides were identified at the metal-ceramic interfaces beyond those present in the oxidized alloys and the opaque porcelain. Similar results were obtained from alloy-ceramic interfaces where there was no prior alloy oxidation. The results indicate the critical role of alloy surface oxides in metal-ceramic bonding and support the chemical bonding mechanism for porcelain adherence.

Journal Article↗

PubMed2001

Intrauterine hypoxia-ischemia alters expression of the NMDA receptor in the young rat brain.

Effects of intrauterine hypoxia-ischemia (HI) on expression of the NMDA receptor subunits as well as on [3H]MK-801 binding of the NMDA receptor were studied in 1-day to 30-day old rat brain. Intrauterine HI conditions were achieved on gestation day 17 by clamping the uterine vasculature for 30 min followed by removal of the clamps to permit reperfusion. As determined by reverse-transcriptase polymerase chain reaction, prenatal HI significantly reduced mRNA expression of the NRI subunit of the NMDA receptor in the hippocampus of 4, 8, and 30-day old rat brains. NR2A and NR2B subunit mRNAs were expressed in the hippocampus and the cortex of both the control and the prenatal HI rat brains. Intrauterine HI did not significantly affect expression of either the NR2A or NR2B subunit mRNA. Consistent with the RT-PCR data, protein expression of the NRI subunit in the hippocampus, but not the cortex, of 21-day old prenatal HI rat brains was significantly decreased as compared to the control rat brain. Intrauterine HI also significantly reduced binding affinity, but not the number of binding sites, of the NMDA receptor to [3H]MK-801, a noncompetitive antagonist of the NMDA receptor, in the hippocampus of 21-day old rat brain. The overall results suggest that prenatal HI-induced reduction of NRI expression and the altered binding ability of the NMDA receptor in the young rat brain may contribute to other long-lasting effects of intrauterine HI that we reported previously.

Aging↗

PubMed2001

Apoptotic response to homoharringtonine in human wt p53 leukemic cells is independent of reactive oxygen species generation and implicates Bax translocation, mitochondrial cytochrome c release and caspase activation.

In the present study, we investigated the in vitro apoptotic response of leukemic cells to the cellular stress induced by homoharringtonine (HHT), a plant alkaloid with antileukemic activity which is currently being tested for treatment of acute and chronic leukemias. A comparison of leukemic cell lines with different p53 gene status revealed a considerably higher sensitivity to HHT-induced apoptosis in the cells with a wt p53, and apoptotic events in wt p53 leukemia cells (MOLT-3 cell line) were studied in more detail. To this end, we examined components of apoptotic cascades including Bax expression and its intracellular localization, changes of mitochondrial membrane potential (MMP), reactive oxygen species (ROS) levels, cytochrome c release from mitochondria and activation of caspases. Bax protein levels did not increase despite an up-regulation of bax at mRNA level. However, Bax translocation from cytosol towards mitochondria was observed. In addition, we observed a release of cytochrome c from the mitochondria, and the localization changes of both Bax and cytochrome c were found already at the early, annexin V-negative stage of HHT-induced apoptosis. HHT-treated MOLT-3 cells revealed loss of MMP as well as activation of caspases demonstrated by DEVD-, IETD- and LEHD-tetrapeptide cleavage activity in the cell lysates. ROS levels only slightly increased in HHT-treated cells and antioxidants did not prevent apoptosis and MMP changes. Therefore, wt p53 leukemic cells respond to HHT-specific cellular stress by induction of ROS-independent apoptotic pathway characterized by translocation of Bax, mitochondrial cytochrome c release and activation of caspases.

Antioxidants↗

PubMed2001

In vitro susceptibility to TRAIL-induced apoptosis of acute leukemia cells in the context of TRAIL receptor gene expression and constitutive NF-kappa B activity.

The TNF-related apoptosis-inducing ligand (TRAIL) is currently under evaluation as a possible (co-)therapeutic in cancer treatment. We therefore examined 129 cell samples from patients with de novo acute leukemia as to their constitutive susceptibility to TRAIL-induced apoptosis In vitro. Only 21 (16%) cell samples revealed at least 10% TRAIL-susceptible cells/sample as detected by flow cytometric annexinV staining after 24 h culture compared with medium control. Precursor B cell ALL samples (11 (27%) of 41) were more TRAIL-susceptible compared with AML (5 (9%) of 54; P < 0.05) but not compared with precursor T cell ALL (5 (15%) of 34; P = 0.20). Furthermore, we examined constitutive mRNA expression levels of TRAIL receptors R1-R4 by semi-quantitative RT-PCR (n = 58). Expression levels were heterogeneous, however, there was no significant correlation between the expression of the signal-transducing receptors (R1, R2) as well as of the decoy receptors (R3, R4) and TRAIL sensitivity in this series. Constitutive NF-kappa B activity has been shown to influence TRAIL susceptibility of leukemic cells. In 39 leukemic cell samples examined, we found a generally high NF-kappa B activity as detected by electrophoretic mobility shift assay which did not differ between TRAIL-susceptible and TRAIL-resistant cases. Finally, 49 acute leukemic cell samples were coincubated with doxorubicin in vitro. Doxorubicin sensitized four of 35 initially TRAIL-resistant samples and augmented TRAIL-induced apoptosis in two of 14 TRAIL-susceptible samples. In summary, constitutive TRAIL susceptibility differs between leukemia subtypes and does not correlate with mRNA expression levels of the TRAIL receptors R1-R4 as well as constitutive NF-kappa B activation status. The observed sensitization of leukemic cells to TRAIL by doxorubicin in vitro indicates that TRAIL should be further evaluated as to its possible role as an in vivo cotherapeutic in acute leukemia.

Acute Disease↗

PubMed2001

Cyclic AMP-dependent protein kinase phosphorylates group III metabotropic glutamate receptors and inhibits their function as presynaptic receptors.

Recent evidence suggests that the functions of presynaptic metabotropic glutamate receptors (mGluRs) are tightly regulated by protein kinases. We previously reported that cAMP-dependent protein kinase (PKA) directly phosphorylates mGluR2 at a single serine residue (Ser843) on the C-terminal tail region of the receptor, and that phosphorylation of this site inhibits coupling of mGluR2 to GTP-binding proteins. This may be the mechanism by which the adenylyl cyclase activator forskolin inhibits presynaptic mGluR2 function at the medial perforant path-dentate gyrus synapse. We now report that PKA also directly phosphorylates several group III mGluRs (mGluR4a, mGluR7a, and mGluR8a), as well as mGluR3 at single conserved serine residues on their C-terminal tails. Furthermore, activation of PKA by forskolin inhibits group III mGluR-mediated responses at glutamatergic synapses in the hippocampus. Interestingly, beta-adrenergic receptor activation was found to mimic the inhibitory effect of forskolin on both group II and III mGluRs. These data suggest that a common PKA-dependent mechanism may be involved in regulating the function of multiple presynaptic group II and group III mGluRs. Such regulation is not limited to the pharmacological activation of adenylyl cyclase but can also be elicited by the stimulation of endogenous G(s)-coupled receptors, such as beta-adrenergic receptors.

Adrenergic beta-Agonists↗

PubMed2001

Focal myelin swellings and tomacula in anti-MAG IgM paraproteinaemic neuropathy: novel teased nerve fiber studies.

Focal myelin swellings and tomacula in teased nerve fibers from a case of IgM anti-myelin-associated glycoprotein (MAG) paraproteinaemic neuropathy were examined using a novel technique. Five different morphologic abnormalities were identified--myelin sheath outfolding, myelin sheath infolding, enlargement of the adaxonal space, myelin degeneration, multiple increased concentric loops--and a combination of these structural abnormalities often occur in association with myelin degeneration. Similar structural changes were found in externally normal segments of teased fibers without evidence of myelin swelling or tomacula from the same case. These structural abnormalities are consistent with a disturbance of the normal adhesion functions of MAG in the maintenance of axon-myelin relationships.

Aged↗

PubMed2001

The use of psychoactive substances among adolescent students in an area in the south-west of China.

AIM: To survey drug/psychoactive substance use among adolescent students in a south-west province of China. DESIGN: A cluster sample was drawn from this province of nine districts or cities. Each district/city provided two schools from grade 11 senior high school, A total of 18 schools were selected randomly. FINDINGS: A total of 2649 students completed this self-report questionnaire, mean age 17.1 +/- 0.9 years. The response rate was 92.7-95.6% for each of the specific substances or drugs. The 'life-time prevalences of regular substance use' (at least 15 times during in any one month) were, in rank order of prevalence: tobacco 6.3%, non-steroid anti-inflammatory drugs (NSAID) 2.9%, alcohol 2.9%, solvent 0.3%, sedative/hypnotic 0.2% and cannabis 0.04%. The life-time prevalences of at least some use were: alcohol 66.1%, NSAID 59.3%, tobacco 27.4%, sedative/hypnotic 5.2%, heroin 3.1%, solvents 2.8%, amphetamine-type stimulants (ATS) 0.7% and cannabis 0.3%. The 'prevalences of current regular use' (at least 15 times in the past month) were: tobacco 4.2%, alcohol 1.6%, NSAID 0.8%, sedative/hypnotic 0.1%, solvents 0.1% and cannabis 0.1%. The 'prevalences of current use at any level' were: alcohol 15.2%, NSAID 9.6%, tobacco 7.1%, sedative/hypnotic 0.5%, solvents 0.4%, cannabis 0.1%, heroin 0.1%, and ATS 0.04%. The median age at onset substance use was between 10.7 and 13.4 9.6%. CONCLUSIONS: Drug misuse has appeared among teenage students in this area. The most widely used substances were alcohol and cigarettes. The rates of solvent, tobacco and alcohol use among males were substantially higher than in females.

Adolescent↗

PubMed2001

[Involvement of mitochondrial membrane potential in the homoharringtonine induced apoptosis of leukemic T-cells].

OBJECTIVE: Investigation of the role of mitochondrial membrane potential (MMP) in the homoharringtonine (HHT)-induced apoptosis. METHODS: Annexin V staining, flow cytometry and confocal laser scan microscopy were used to observe the relationship between Bax, cytochrome C and MMP in the HHT-induced apoptosis of leukemic T lymphocytic line Molt-3. RESULTS: The induction of apoptosis by HHT resulted in the translocation of Bax from cytosol to mitochondrial membrane and the decrease of cellular MMP, followed by the release of cytochrome C from mitochondria to cytosol. CONCLUSIONS: Changes of mitochondrial membrane potential might play a critical role in the HHT-induced apoptosis of leukemic T-cells.

Apoptosis↗

PubMed2001

[Aminopeptidase inhibitor Bestatin induces HL-60 cell apoptosis through activating caspase 3].

OBJECTIVE: To study the variation and significance of caspase 3 activity in the process of amino-peptidase inhibitor--bestatin (BS) inducing human leukemic cell apoptosis. METHODS: Cell apoptosis was evaluated by light microscopy, TUNEL labeling and flow cytometry (FCM). Caspase 3 activity was detected by colorimetry. The mitochondrial transmembrane potentials (DeltaPsi(m)) were detected by Rhodamine123 staining. RESULTS: The apoptotic morphology, apoptotic peak on FCM and positive Annexin V(FITC) on cell membrane showed that BS could induce HL-60 cell apoptosis in a dose- and time-dependent manner. Caspase 3 activity was significantly higher in the apoptotic cells than in control cells. The apoptosis induced by BS was inhibited by AC-DEVD-CHO. The DeltaPsi(m) of cells treated with BS declined. CONCLUSION: BS induces apoptosis of human acute leukemic cells through activation of caspase 3.

Aminopeptidases↗

PubMed2001

[Applied study on computerized quantitative facial nerve functional estimating system].

OBJECTIVE: To prove the clinical applied value of quantitative facial nerve functional estimating system (QFES) by comparing the estimating results of the computerized QFES with manual QFES. METHODS: 50 volunteers with normal facial nerve function were used. Every volunteer was taken 7 photos with digital camera, which including static, and forehead wrinkle, eyebrows knit tight, eyes closed tight, nose wrinkle, smile, pout and mouth open to maximal seven expressions. The facial nerve function was estimated by computerized QFES, and the estimating results were compared with manual estimations by QFES in our study during 1991-1994. RESULTS: All estimating indices in this study were in the range of 0.70-0.90 as well as the manual estimating results. However, the stand deviation in this study showed significantly smaller than the previous study. CONCLUSIONS: QFES is an ideal quantitative facial nerve functional estimating system, after computerization the efficiency is increased and the systemic error is decreased. It will benefit to apply computerized QFES in the facial nerve function estimating fields.

Diagnosis, Computer-Assisted↗

PubMed2001

[A comparative morphometrical study on development of anterior septonasal accessory organs in the cleft palate mice].

OBJECTIVE: The aim of this study was to investigate whether cleft palate is associated with the mal-development of the vomer, Jacobson's organ and para-septal cartilage. METHODS: Sixty craniomaxillary complexes, which came from 30 pairs of trisomy 18 with cleft palate (Ts + CP) and euploid mouse fetuses were precisely orientated in the coronal plane, and serially sectioned at 7 microns thickness. With the aid of computer imaging analysis system the vomer in coronal and sagittal direction, Jacobson's organ and para-septal cartilage in the boundary plane of the primary and secondary palate were measured and compared quantitatively. RESULTS: The hard tissues (including the vomer and para-septal cartilage) of the anterior nasal septum in Ts + CP foetuses show serious developmental hypoplasia, however, the Jacobson's organ had no significant different between the cleft and non-cleft samples. CONCLUSION: Development hypoplasia of these hard tissues may be more closely related with the autosomal trisomic condition more than the cleft palate.

Animals↗

PubMed2001

[Sequences of 5S-rRNA gene spacer region and comparison of alkaloid content in Fritillaria thunbergii from different habitats].

OBJECTIVE: To study the genetic basis of the formation of indigenous Chinese medicine materials. METHODS: The 5S-rRNA gene spacer regions in F. thunbergii from different habitats were amplified with AS and AS-1 as primers, and then sequenced. Total alkaloid contents were assayed by acid dye colorimetry, and 2 main alkaloid contents were assayed by pre-column derivatization and gas chromatographic method. RESULT: The sequenues of 5S-rRNA gene spacer regions in F. thunbergii from different habitats were same, and the length of them was 588 bp. They had same content total alkaloid. The results of gas chromatography showed that they had same kinds of monomer alkaloids, but the contents of different monomer alkaloids were different. CONCLUSION: The difference of alkaloid content in F. thunbergii from various habitats isn't resulted from base sequence variation, but from microenvironment.

Alkaloids↗

PubMed2001

National survey on prevalence of cancer pain.

OBJECTIVE: To collect nationwide basic data about cancer related pain. METHODS: Sixty cancer patients in each province were randomly selected to participate in this survey. The subjects represented all stages of cancer, tumor sites, and different demographic characteristics. Two self-designed structured questionnaires including reasons, types of pain and pain management were used by patients and physicians respectively. Subjects were asked to report whether he/she had experienced any type of cancer related pain and filled out the equivalent questionnaire. The severity of pain was assessed by using "visual analogue scale". Original data input and analysis were using EPI-INFO software package. RESULTS: The result showed that 61.6% (958/1555) of patients had different types of cancer related pain. Majority of pain (85.1%) were caused by advanced cancer. The major reasons (64.4%) for poor management or impedimental factors of pain care are due to patient including over-concern on opioid analgesic addiction, reluctance to report pain or refused to use opioid analgesic until at times when pain is intolerable; 26.8% belonged to physician's reasons including fear to cause addiction on opioid and lack of knowledge about cancer pain management; 16.2% are due to lack of different kinds of opioid analgesic for use and 16.1% belonged to drug regulation. CONCLUSIONS: The results showed that majority of patients (61.6%) had different types of cancer related pain. In most of patients, cancer pain was relieved when they were treated. The major reason for under-treatment or impeded factors for effective relief of cancer pain was fear of opioid addiction by both medical professionals and patients.

Adolescent↗

PubMed2000

Resistance to TNF-induced cytotoxicity correlates with an abnormal cleavage of cytosolic phospholipase A2.

To investigate the mechanism underlying the absence of arachidonic acid (AA) release by TNF in TNF-resistant cells, we first performed comparative analysis of phospholipid pools in both TNF-sensitive (MCF7) and their equivalent resistant cells (C1001). Quantification and incorporation studies of [(3)H]AA indicated that TNF-resistant cells were not depleted in AA. Furthermore, distribution of this fatty acid in different phospholipid pools was similar in both sensitive cells and their resistant counterparts, ruling out a defect in phospholipid pools. Since phospholipase A(2) (PLA(2)) are the main enzymes releasing free AA, we investigated their relative contribution in the acquisition of cell resistance to TNF-induced cell death and AA release. For this purpose, we used two PLA(2) inhibitors, methylarachidonyl fluorophosphate (MAFP) and bromoenol lactone (BEL), which selectively and irreversibly inhibit the cytosolic PLA(2) (cPLA(2)) and the Ca(2+)-independent PLA(2), respectively. Although a significant inhibitory effect of MAFP on both TNF-induced AA release and PLA(2) activity in MCF7 was observed, BEL had no effect. The inhibitory effect of MAFP on cPLA(2) activity correlated with an inhibition of TNF-induced cell death. Western blot analysis revealed that TNF induced a differential cleavage of cPLA(2) in TNF-sensitive vs TNF-resistant cells. Although the p70 (70-kDa) form of cPLA(2) was specifically increased in TNF-sensitive cells, a cleaved form, p50 (50 kDa), was selectively observed in TNF-resistant C1001 cells in the presence or absence of TNF. These findings suggest that the acquisition of cell resistance to this cytokine may involve an abnormal cPLA(2) cleavage.

Arachidonic Acid↗

PubMed2000

Effects of lipopolysaccharide on oligodendrocyte progenitor cells are mediated by astrocytes and microglia.

Oligodendrocytes are the primary cells injured in periventricular leukomalacia (PVL), a predominant form of brain white matter lesion in preterm infants. To explore the possible linkage between white matter injury and maternal infection, purified rat O-2A progenitor (Oligodendrocyte-type 2 astrocyte progenitor) cell cultures were used as a model in studying the effects of lipopolysaccharide (LPS), an endotoxin, on survival and differentiation of oligodendrocytes and the involvement of other glial cells in the effects of LPS. O-2A progenitor cells were cultured from optic nerves of 7-day-old rat pups in a chemically defined medium (CDM). Astrocyte and microglia cell cultures were prepared from the cortex of 1-day-old rat brains in the CDM. Direct treatment of LPS (1 microg/ml) to O-2A cells had no effect on viability or differentiation of these cells. When O-2A progenitor cells were cultured in the conditioned medium obtained from either astrocyte or microglial cell cultures for 48 hr, survival rate and differentiation of O-2A cells into mature oligodendrocytes were greatly enhanced as measured by the MTT assay and immunocytochemistry. The conditioned medium obtained from astrocytes or microglia treated with LPS for 48 hr, however, failed to show such a promotional effect on viability and differentiation of O-2A cells. When 5 microg/ml LPS was used to stimulate astrocytes or microglia, the conditioned medium from these glial cell cultures caused O-2A cell injury. The overall results indicate that astrocytes and microglia may promote viability and differentiation of O-2A progenitor cells under physiological conditions, but they may also mediate cytotoxic effects of LPS on oligodendrocytes under an infectious disease biochemical environment.

Animals↗

PubMed2000

Freshly isolated astrocyte (FIA) preparations: a useful single cell system for studying astrocyte properties.

Astrocytes are cell constituents of the mammalian CNS whose intricate relationships with neurons, blood vessels and meninges in situ are well documented. These relationships and their complex morphologies imply numerous functions. Over the past quarter century or so, however, the main experimental basis for determining which roles are likely have been derived from studies on primary astrocyte cultures, usually prepared from neonatal rodent brains. We list a number of examples where these cultures have shown quantitative and qualitative differences from the properties exhibited by astrocytes in situ. The absence of an adequate reliable database makes proposals of likely hypotheses of astrocyte function difficult to formulate. In this article we describe representative studies from our laboratory showing that freshly isolated astrocytes (FIAs), can be used to determine the properties of astrocytes that seem more in concordance with the properties exhibited in situ. Although the cells are most easily isolated from < or =15 day old rat hippocampi they can be isolated from up to 30 day old rats. The examples we describe are that several different types of K(+) currents can be determined by patch clamp electrophysiology, of all the mGluRs only mGluR3 and 5 were detected by single cell RT-PCR, and that single cell Ca(2+) imaging shows that the mGluR5 receptor is functional. It was found that the frequency of cells expressing mGluR5 declines with the age of the animal with the mGluR5b type splice variant replacing the mGluR5a type, as occurs in the intact brain. It is concluded that FIAs can be used to determine the individual characteristics of astrocytes and their properties without the problems of indirect effects inherent in a heterogeneous system such as the slice, and without the problem of cultures unpredictably reflecting the in situ state. The FIAs obviously cannot be used to study interactions of astrocytes with the other CNS components but we propose that they will provide a good database on which hypotheses regarding such interactions can be tested in slices. FIAs can also be isolated from brain slices or intact brain after various pharmacological or electrophysiological perturbations to determine the changes in astrocyte properties that correlate with the perturbations.

Animals↗