Biomedical subjects

M Huleihel

Publications and source records attributed to M Huleihel.

At least 19 recordsLinked to original sources

PubMed2006

Normal aging of offspring mice of mothers with induced inflammation during pregnancy.

Intrauterine inflammation is a major risk for offspring neurodevelopmental brain damage and may result in cognitive limitations and poor cognitive and perceptual outcomes. In the present study we tested the possibility that prenatal exposure to a high level of inflammatory factors may increase the risk for neurodegeneration in aging. The effect of systemic maternal inflammation (MI), induced by lipopolysaccharide (LPS) on offspring brain aging, was examined in 8 month old (adult) and 20 month old (aged) offspring mice. A significant effect of age was found in the distance and velocity of exploration in the open field in both groups. In addition, MI aged offspring covered longer distances and enter frequently to the center of the field compared with the aged control group. Although only little difference was found in the aged MI offspring compared with the control offspring, the overall profile of behavior of these mice differs from that of the control group, as detected by clustering analysis. The expression of the death-associated protein FAS-ligand and the amount of apoptotic cell death were examined in the brains of aged offspring. Similar levels of FAS-ligand expression and parallel density of apoptotic cells were detected in the brains of aged mice of control and MI groups. Altogether, moderate systemic MI was not found to increase the risk for cell death in the aged offspring; limited effect was found in mice profile of behavior.

Age Factors↗

PubMed2005

Remodeling of hippocampal GABAergic system in adult offspring after maternal hypoxia and magnesium sulfate load: immunohistochemical study.

A strong relationship between hypoxia and fetal brain damage has been described. Specific susceptibility of the GABAergic neurons to these conditions may be crucial to the damage induced. We have previously shown, in a mouse model, that maternal pretreatment with magnesium sulfate (Mg) partially prevented the behavioral consequences of maternal hypoxia in the adult offspring. Here, we tested the effect of maternal hypoxia and maternal Mg load on the GABAergic system of 8-month-old offspring. The immunoreactivity (IR) of several proteins expressed in GABAergic neurons and inhibitory synapses was analyzed in the following regions of the adult offspring brain: hippocampus, cortical M1, caudate putamen, and lateral globus pallidus. Maternal hypoxia reduced the density of parvalbumin (PV)-IR neurons in the hippocampus. The density of PV-IR and calbindin (CB)-IR neurons was also reduced in the deep and superficial layers of the M1. Maternal pretreatment with Mg had a prophylactic action in the superficial, but not the deep, layers of M1. Also, in offspring from the maternal hypoxia group, the vesicular GABA transporter (VGAT)-IR was enhanced in the hippocampal CA1 and hilus regions. No effect of maternal hypoxia on VGAT-IR was observed in the M1. However, maternal pretreatment with Mg enhanced VGAT-IR and glutamate decarboxylase-IR in the deep layers of the M1. In the globus pallidus, maternal hypoxia enhanced CB-IR, which was prevented by maternal pretreatment with Mg. In conclusion, maternal hypoxia induced a loss of PV-IR and CB-IR neurons; maternal pretreatment with Mg partially protected these neuron populations. An increase in proteins of inhibitory synapses, observed under hypoxic conditions in several brain regions, may be a result of some compensatory mechanism.

Animals↗

PubMed2005

Specific neurodevelopmental damage in mice offspring following maternal inflammation during pregnancy.

Intrauterine inflammation is a major risk for offspring neurodevelopmental brain damage and may result in cognitive limitations and poor cognitive and perceptual outcomes. Pro-inflammatory cytokines, stimulated during inflammatory response, have a pleotrophic effect on neurons and glia cells. They act in a dose-dependent manner, activate cell-death pathways and also act as trophic factors. In the present study, we have examined in mice the effect of short, systemic maternal inflammation on fetal brain development. Maternal inflammation, induced by lipopolysaccharide (LPS) at gestation day 17, did not affect morphogenic parameters and reflex development during the first month of life. However, maternal inflammation specifically increased the number of pyramidal and granular cells in the hippocampus, as well as the shrinkage of pyramidal cells, but not of the granular cells. No additional major morphological differences were observed in the cerebral cortex or cerebellum. In accordance with the morphological effects, maternal inflammation specifically impaired distinct forms of learning and memory, but not motor function or exploration in the adult offspring. The specific deficiency observed, following maternal inflammation, may suggest particular sensitivity of the hippocampus and other associated brain regions to inflammatory factors during late embryonic development.

Age Factors↗

PubMed2004

Maternal hypoxia during pregnancy induces fetal neurodevelopmental brain damage: partial protection by magnesium sulfate.

Fetal low brain oxygenation may be an outcome of maternal complications during pregnancy and is associated with increased risk of cerebral palsy and periventricular leukomalacia in newborns. One treatment used for prevention of fetal brain damage is maternal treatment with MgSO(4). Although this treatment is indicated to reduce the risk of cerebral palsy in newborns, its use remains controversial. We have shown previously that pretreatment with MgSO(4) in a mouse model of maternal hypoxia prevented a delay in the development of motor reflexes induced by hypoxia. We demonstrate here that pretreatment with MgSO(4) reduces hypoxia-induced motor disabilities in adult offspring. This effect is associated with histologic protection of the Purkinje cells in the cerebellum and stabilization of brain-derived neurotrophic factor (BDNF) levels in the cerebellum. MgSO(4) did not prevent the reduction in cerebral cortex cell density and cell size induced by maternal hypoxia, however, nor did it interfere with the modulation of BDNF and nerve growth factor (NGF) expression in the cerebral cortex. MgSO(4) pretreatment also prevented the impairment of short-term memory (30 min, P < 0.05) but not long-term memory (7 days). Nevertheless, maternal pretreatment with MgSO(4) reduced CA1 cell layer width and induced alterations in both NGF and BDNF in the hippocampus. These results support the prophylactic effect of MgSO(4) against motor disabilities; however, they may also indicate possible harmful effects on the cerebral cortex and hippocampus.

Age Factors↗

PubMed2004

Involvement of tumor necrosis factor alpha in hippocampal development and function.

Tumor necrosis factor alpha (TNFalpha) is a cytokine produced mainly by cells of the immune system. It is also expressed by brain neurons and glia. The physiological role of TNFalpha in the brain is not yet fully clear. Using TNFalpha-deficient mice, we have examined its role in hippocampal development and function. We report here that TNFalpha is involved in the regulation of morphological development in the hippocampus. TNFalpha-deficient mice exhibited an accelerated maturation of the dentate gyrus region and smaller dendritic trees in CA1 and CA3 regions in young mouse. In addition to its involvement in hippocampal morphogenesis, TNFalpha deficiency specifically improved performance of affected mice in behavioral tasks related to spatial memory. Moreover, lack of TNFalpha increased the expression of nerve growth factor (NGF), but not brain-derived neurotrophic factor (BDNF), following performance of the learning task. Our results suggest that TNFalpha actively influences hippocampal development and function. In adult mice, TNFalpha may interfere with memory consolidation, perhaps by regulating NGF levels.

Aging↗

PubMed2004

Characteristic absorbance of nucleic acids in the Mid-IR region as possible common biomarkers for diagnosis of malignancy.

FTIR spectroscopy has been extensively used to understand the differences between normal and malignant cells and tissues. In the present study, FTIR microspectroscopy was performed on biopsies to evaluate parameters deduced from changes in nucleic acid absorbance monitored at various characteristic wavenumbers in the Mid-IR region. The data showed that there were differences in the spectra of normal and malignant tissues from several organs such as colon, cervix, skin and blood with respect to absorbance due to nucleic acids. Similar results were observed in the case of cell lines that were transformed to induce carcinogenesis. Of the several ratios examined for consistency in differentiating cancer and normal tissues, the I(996 cm(-1))/I(966 cm(-1)) showed promise as a distinguishing parameter and was comparable to the I(1121 cm(-1))/I(1020 cm(-1)) ratio reported in many earlier studies. The absorbance of nucleic acids is presented with an emphasis on the application of FTIR microspectroscopy for diagnosis of malignancy. Our results indicate that usage of nucleic acid absorbance yield statistically significant parameters, which could differentiate normal and cancerous tissues.

Animals↗

PubMed2003

TPA activates p21WAF-1 promoter in human T-cells through its second most upstream Sp1 site.

The p21(WAF-1) promoter contains binding sites for a number of transcription factors which mediate its activation by a variety of external signals. Moreover, it has been reported that the transcription factors involved in p21(WAF-1) activation by certain signaling factors, like the phorbol ester TPA, may vary in different cell types. We were interested in elucidating the mechanism of p21(WAF-1) activation by TPA in human T-cells, since this activation could explain the antagonistic effect of PKC on apoptosis induction in these cells noted in our previous studies. Using the Jurkat human T-cells we found that TPA activated p21(WAF-1) expression by a PKC-dependent mechanism and that out of six Sp1 binding sites residing in its promoter the second most upstream one was critically essential for this activation. Since p21(WAF-1) is known to inhibit the onset of apoptosis, its PKC-dependent activation may likely account for the PKC antagonistic effect on apoptosis induction in these cells.

Binding Sites↗

PubMed2003

[Matrix metalloproteinases and their tissue inhibitors in malignancies of the female genital tract].

The matrix metalloproteinases (MMPs) are a family of proteolytic enzymes, which are capable of degradation of the proteins composing the extracellular matrix and basement membrane. Their proteolytic activity depends on their binding to metal Zinc and is controlled by tissue inhibitors of MMP (TIMPs). Degradation of the extracellular matrix and basement membrane is an important component of the process of tumor invasiveness, progression, angiogenesis and metastatic spread. Since MMPs may serve as markers of tumor behavior and as predictors of survival and since synthetic inhibitors of MMP may have a place in the treatment of cancer, researching MMPs and their tissue inhibitors in malignant diseases has attracted growing attention. Studies on MMPs and their tissue inhibitors in malignancies of the female genital tract have shown the following: 1) In ovarian carcinoma and cervical carcinoma, over-expression of MMP-2 and MMP-9 is associated with invasiveness, metastatic spread and poor prognosis; 2) In endometrial carcinoma, MMP-7 (matrilysin) is the main MMP associated with invasiveness, metastatic spread and poor prognosis; 3) In cervical intra-epithelial neoplasia (CIN), measuring MMP-2 can assist in identifying high-risk for progression CIN I and CIN II; 4). In vulvar squamous cell carcinoma, over-expression of MMP-13 is associated with invasiveness, metastatic spread and poor prognosis. It is speculated that using synthetic drugs that inhibit MMPs in combination with conventional chemotherapy may contribute to the improvement of treatment results in cancer patients.

Biomarkers, Tumor↗

PubMed2002

Novel methodology for the follow-up of acute lymphoblastic leukemia using FTIR microspectroscopy.

In this report, we present a novel spectroscopic method of follow-up during chemotherapy treatment for B- and T-cell childhood leukemia patients. We isolated peripheral lymphocytes from blood drawn from patients before and after the chemotherapy and collected Microscopic FTIR (FTIR-MC) spectra of the isolated lymphocytes. Our results showed that nucleic acids content decreased in both types of patients. Changes in phospholipids and proteins level could be observed. The overall effects of drugs administered to the patients can be understood at the molecular level using FTIR-MC and these results are expected to stimulate wider applications of spectroscopy in leukemia research.

Adolescent↗

PubMed2001

Effect of propolis extract on malignant cell transformation by moloney murine sarcoma virus.

An aqueous extract of propolis was found to significantly inhibit NIH/3T3 cell malignant transformation by Moloney murine sarcoma virus (MuSV-124). The inhibitory effect of propolis extract was most effective when it was added 2 h before infection or at the time of infection. The continuous presence of propolis extract in the culture medium was essential for full prevention of malignant cell transformation. When treatment with propolis extract was terminated, five to ten days post-infection, there was a significant recovery in cell transformation. These results suggest that propolis extract inhibits a late step after provirus integration into the host genome. Addition of propolis extract after infection with MuSV significantly inhibited cell transformation. The inhibitory effect of propolis appeared to be the result of the inhibition of primary--not secondary--infections, since MuSV-124 yields a virus-nonproducing infection.

3T3 Cells↗

PubMed2001

Increased production of tumor necrosis factor-alpha TNF-alpha by IUGR human placentae.

OBJECTIVE: To evaluate the effect of pathological placental conditions such as intrauterine growth restriction (IUGR) or exposure to angiotensin II (AII) on TNF-alpha secretion in the vasculature of isolated human placental cotyledons. STUDY DESIGN: Isolated placental cotyledons from 10 normal and four intrauterine growth restricted fetuses were dually perfused. Perfusate samples from the fetal circulation were collected every 30 min during 120 min. TNF-alpha levels in the fetal-placental perfusate were evaluated using specific commercial ELISA kits. In three additional normal placentae, bolus injections of angiotensin II (10(-9)-10(-4) mol/l) were given into the fetal-placental circulation and perfusate samples were collected. Statistical significance of difference TNF-alpha levels between different conditions was determined by analysis of variance (ANOVA) and paired t-test. RESULTS: TNF-alpha levels were significantly higher in the perfusate of IUGR placentae as compared with normal placentae after 120 min of perfusion (mean 410+/-121 vs. 39+/-14 pg/ml, P=0.005). There was a significant dose-dependent increase in TNF-alpha levels in the placental perfusate after a bolus injection of AII 66 pg/ml with AII 10(-9) mol/l vs. 97 pg/ml with AII 10(-5) mol/l (P=0.004), respectively. CONCLUSIONS: Placental pathology related to condition IUGR might induce the secretion of proinflammatory cytokines such as TNF-alpha, which may enhance the vasoconstriction of the fetal placental vascular bed.

Angiotensin II↗

PubMed2001

Indomethacin activity in the fetal vasculature of normal and meconium exposed human placentae.

OBJECTIVE: To study the effect of indomethacin on the vasculature of isolated perfused human placental cotyledon in normal and meconium pretreated placentae. STUDY DESIGN: Isolated placental cotyledons were dually perfused and fetal perfusion pressure was used as an index of vascular resistance. Meconium-stained amniotic fluid (MSAF) was collected from patients after artificial rupture of membranes, diluted 1:2, 1:4, 1:16 and 1:32 and a spectrophotometric determination of meconium concentration in amniotic fluid was performed. Only MSAF with an optical density of 20.0 units per gram was used in this study. In five placentae, the effect of indomethacin (100 microg/ml continuous perfusion from the fetal site) on basal pressure of the fetal-placental vasculature was established. In five more placentae, the effect of indomethacin on MSAF-induced vasoconstriction was established when a bolus injections of 1 ml MSAF was made into the fetal circulation. The statistical significance of response to MSAF injection was determined by paired t-test and ANOVA repeated measurements. RESULTS: A significant vasoconstrictor response to MSAF was achieved in normal placentae. Bolus injections of MSAF into the fetal circulation resulted in a significant increase in perfusion pressure (P=0.0026). Indomethacin was capable of significantly reducing the basal perfusion pressure (P=0.03). Significant attenuation of vasoconstrictor response to MSAF occurred in the presence of indomethacin (P=0.0016). CONCLUSION: Indomethacin causes a significant reduction in basal pressure of fetal placental vasculature in the human placental circulation in vitro and is capable of attenuating the vasoconstrictory activity of MSAF. The mechanism of such activity may be explained partially by the inhibitory effect of indomethacin on the PG-mediated pathways.

Amniotic Fluid↗

PubMed2001

Immunohistochemical staining of IL-1 alpha and IL-1 receptor antagonist but not IL-1 beta in cultures of sertoli cells.

The interleukin-1 (IL-1) system has been suggested to be involved in the cell cell cross talk within the testis. To identify a testicular cell source of IL-1 alpha, IL-1 beta and IL-1 receptor antagonist (IL-1ra), immature mouse Sertoli cells were isolated, purified, cultured and examined for the cellular compartment localization of these cytokines by immunohistochemical staining. Our results show that both Germ cells and Sertoli cells in unpurified Sertoli cell cultures (before hypotonic shock) and purified culture of Sertoli cells (after hypotonic shock) were stained for IL-1 alpha. The levels of this cytokine were increased in Sertoli cells when the purified cultures were stimulated with lipopolysaccharide (LPS) (5 microg/mL). However, we could not identify a positive staining for IL-1 beta when Sertoli cell cultures were stained for this cytokine, even after stimulation with various concentrations of LPS (0.1-10 microg/mL). On the other hand, immunohistochemical staining of isolated Sertoli cells without treatment with hypotonic shock (cultures containing Sertoli cells and Germ cells) for IL-1ra showed constitutive positive staining of both cell types (Sertoli cells and Germ cells). Our results, using immunohistochemical staining, may indicate the different expression of IL-1 alpha, IL-1 beta and IL-1ra in Sertoli cells. These results may suggest the involvement of IL-1 system in the autocrine and paracrine regulation of testicular cell functions.

Animals↗

PubMed2001

Albumin-dependent digoxin transfer in isolated perfused human placenta.

OBJECTIVE: To determine the effects of albumin (BSA) concentration in perfusion medium on digoxin transfer in isolated perfused human placental cotyledon. STUDY DESIGN: Isolated placental cotyledons from 13 normal human placentas were dually perfused after cannulating artery and vein of the chorionic plate and piercing 4 catheters through the corresponding basal plate with M199 medium enriched with BSA and glucose. Flow rates were 12 and 6 ml/min in the maternal and fetal circuits, respectively. Digoxin was added to the maternal reservoir at a final concentration of 5.51 +/- 1.00 ng/ml. BSA in maternal and fetal perfusate was kept at 3 concentrations: 1, 3 and 5 mg/ml (Groups I, II, III). Transplacental passage of digoxin was calculated from repeated fetal and maternal perfusate samples collected over 3 hours in the 3 groups. Digoxin levels were measured by FPIA (TDx, Abbott). RESULTS: There was no transfer of digoxin from the maternal to fetal compartment when the concentration of BSA was 1 mg/ml. Increasing the concentration of BSA led to a substantial increase in the transfer of digoxin to the fetal compartment. Steady state levels of digoxin in the fetal compartment were 0.61 +/- 0.19 ng/ml at 3 mg/ml of BSA. CONCLUSION: Maternal and fetal serum concentration of BSA affect digoxin transfer in isolated perfused human placentas. Three mg/ml are considered to be the optimal albumin concentration.

Analysis of Variance↗

PubMed2001

Fetal death associated with asymptomatic intrauterine Candida albicans infection and a retained intrauterine contraceptive device.

We present a case of intrauterine fetal death at 18 weeks of gestation associated with a retained intrauterine contraceptive device and asymptomatic intraamniotic and fetal infection by Candida albicans. The infection was verified by histopathologic examination of the placenta and umbilical cord, growth of C. albicans in samples of amniotic fluid and the presence of high levels of IL-6 in the amniotic fluid.

Adult↗

PubMed2000

Production of interleukin-1-like molecules by human sperm cells.

OBJECTIVE: To characterize and localize interleukin (IL)-1alpha and IL-1beta in human sperm cells. DESIGN: Prospective and comparative study. SETTING: Andrology clinic of a university hospital. PATIENT(S): Two groups of normogonadotropic men: 17 fertile men (donors with proved fertility) and 8 oligoteratoasthenospermic infertile men. INTERVENTION(S): None. MAIN OTUCOME MEASURE(s): Evaluation of IL-1alpha and IL-1beta levels and expression in sperm cells by immunohistochemical staining, immunoassay, and Western blot analysis. RESULT(S): Both types of IL-1-like molecules (IL-1alpha and IL-1beta) were expressed in the tail, neck, and head of sperm cells of fertile men and patients with oligoteratoasthenospermia. Swim-up sperm cells from fertile men and patients with oligoteratoasthenospermia secreted similar levels of IL-1-like molecules. The levels of IL-1beta-like molecules were higher than those of IL-1alpha-like molecules in both groups. The expressed IL-1-like molecules were characterized by the presence a 60-kd protein for both IL-1alpha-like and IL-1beta-like molecules. In some samples of both fertile men and infertile men with oligoteratoasthenospermia, 17-kd, 33-kd, and 45-kd IL-1beta-like molecules were detected. Impairment of sperm function, such as decreased sperm count and motility and/or impaired morphology, was not related to the capacity of sperm cells to produce IL-1-like molecules. CONCLUSION(S): IL-1 molecules originating in sperm cells may play a role in the physiologic functions of sperm cells (autocrine effect) and/or in cell-cell interactions within the testis (paracrine effect).

Blotting, Western↗

PubMed2000

Induction of interleukin-1alpha production in murine Sertoli cells by interleukin-1.

In the present study we examined the involvement of interleukin (IL)-1alpha, -1beta, FSH, and lipopolysaccharide (LPS) in the regulation of IL-1alpha and -1beta production by Sertoli cells under in vitro conditions. Sertoli cell cultures from immature mice produced constitutively basal levels of intracellular IL-1alpha. Stimulation of Sertoli cell cultures with LPS (5 microgram/ml) resulted in a maximal production of intracellular IL-1alpha 2 h after the stimulation. Thereafter, these levels decreased but remained significantly higher within 24 h after stimulation than those in control cultures. The effect of LPS on IL-1alpha production was dose dependent. FSH did not show any effect on intracellular IL-1alpha production by Sertoli cells. IL-1alpha could not be detected in supernatants of unstimulated or stimulated Sertoli cell cultures. Sertoli cell cultures stimulated with recombinant IL-1alpha induced optimal intracellular levels of IL-1alpha within 2 h of stimulation. These levels remained high 24 h after stimulation. However, stimulation of Sertoli cell cultures with IL-1beta induced a peak of IL-1alpha production 8 h after stimulation. These levels decreased 24 h after the stimulation but were still found to be significantly higher than those in control cultures. The addition of IL-1 receptor antagonist (IL-1ra) to Sertoli cell cultures did not significantly alter their capacity to produce IL-1alpha. However, the stimulatory effects of recombinant IL-1alpha on IL-1alpha production by Sertoli cell cultures were reversed by the concomitant addition of recombinant IL-1ra. No immunoreactive IL-1beta could be detected in lysates or conditioned media of immature murine Sertoli cells under any of the stimulatory conditions outlined. Our results may suggest the involvement of physiological (IL-1) and pathophysiological factors (LPS) in the regulation of spermatogenesis and spermiogenesis processes and male fertility.

Animals↗

PubMed2000

Involvement of serum and lipopolysaccharide in the production of interleukin-1- and interleukin-6-like molecules by human sperm cells.

PROBLEM: To examine the capacity of sperm cells from fertile and infertile men to secrete interleukin (IL)-6, and the involvement of serum factors and lipopolysaccharide (LPS) in the regulation of IL-6 and IL-1 production by sperm cells. METHODS: Swim-up sperm cells from fertile (donors) and oligoteratoasthenospermic (OTA)-infertile men were incubated with or without 5% fetal calf serum (FCS) and LPS (10 microg/mL) for 2-24 hr. After incubation, IL-6 and IL-1 bioactivities were measured in supernatants and lysates by specific bioassays (B9 cell proliferation assay and 1A-5 system, respectively). RESULTS: IL-6- and IL-1-like activities were observed to be produced by swim-up sperm cells from both study groups. Stimulation of swim-up sperm cells with either LPS or FCS or both together did not affect their capacity to produce IL-1. However, LPS, but not serum increased the secretion levels of IL-6 by swim-up sperm cells. CONCLUSIONS: Swim-up sperm cells from both study groups constitutively produce IL-6 and IL-1, and serum components did not affect this capacity. However, LPS was shown to increase the capacity of swim-up sperm cells of both study groups to secrete IL-6, but not IL-1. Cytokines may be involved in the physiology and pathophysiology of sperm functions and, thus, may affect male fertility.

Animals↗