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M Strauss

Publications and source records attributed to M Strauss.

At least 55 records · Page 3Linked to original sources
PubMed2000

Salivary gland ultrastructural alterations in mice inoculated with Tityus discrepans (Buthidae) venom.

In this work we have studied the possible relationships between clinical manifestations such as sialorrhea, appearing in response to the toxic aggression by Tityus discrepans venom, and the alterations or changes at cellular or subcellular levels in sub-maxillary salivary glands in the murine model. To evaluate salivary gland subcellular response to Tityus discrepans venom, male C57/Bl adult mice were randomised into two groups: a group of mice were intraperitoneally injected with Tityus discrepans venom at a dose of 5 mg/Kg of weight and controls received saline solution. In the salivary glands from the envenomed animals sub-cellular changes such as hyperchromatic nucleus, swollen rough endoplasmic reticulum, granules of different electron density, some of them practically transparent, nucleolus of low density and big size and electron-transparent cisterns were observed. Capillary wall was augmented in certain areas and thin in others. Endothelial cell infolding to the lumen was seen. The distribution of the vesicles and its density varied. Macrophages and plasmocytes were observed next to the damaged capillaries. Different electron density of cytoplasm was noticed. In conclusion, we suggest that sialorrhea is determined by salivary gland damage produced by toxins present in this venom.

Animals↗
PubMed2000

Cardiac ultrastructural alterations in mice inoculated with Tityus discrepans (Buthidae) venom.

In this work we have studied the cardiologic clinical manifestations appearing in response to toxic aggression by Tityus discrepans venom, such as hypertension, hypotension, tachycardia and pulmonary acute oedema. These depend on changes in the organisation of cellular and subcellular components of cardiac tissues and probably correspond with the damage found in envenomed humans. To evaluate cardiac tissue subcellular response to Tityus discrepans venom, male C57/B1 adult mice were randomised into two groups: envenomed mice were intraperitoneally injected at a dose of 5 mg/Kg of body weight and controls received saline solution. Samples from cardiac tissue were prepared for electron microscopy study and observed in a Hitachi-300. The most relevant cardiac ultrastructural findings in this model showed diffuse disarray of the myofibrils and abnormal pattern of the bands in the sarcomera, contractile element disorganisation, degeneration of fibres and loss of the characteristic sarcomeric structure given the appearance of a lax tissue. One of the most prominent features was the presence of a remarkable perinuclear oedema and the perinuclear cistern exhibited indentations over its whole arrangement. The vascular endothelium of the microvessels exhibited alterations with evident cytoplasmic projections toward the lumen of the vessel. Mitochondria presented a condensed conformation. All findings were degenerative signs of the contractile apparatus. We suggest that any cardiac tissue damage produced by toxins present in this venom are responsible for some of the clinical manifestations in envenomed animals and patients.

Animals↗
PubMed1999

Nonphysiological overexpression of low-density lipoprotein receptors causes pathological intracellular lipid accumulation and the formation of cholesterol and cholesteryl ester crystals in vitro.

Recent therapeutic strategies for the treatment of familial hypercholesterolemia have been based on liver-directed gene transfer of a functional low-density lipoprotein (LDL) receptor cDNA under control of viral or strong housekeeping promoters. Strong viral promoters including cytomegalovirus, Rous sarcoma virus, and simian virus 40 promoters are commonly employed to reach significant physiological effects. These promoters mediate constitutive and nonphysiological overexpression in every transduced cell, while the endogenous LDL receptor expression is controlled by a complex feedback mechanism based on intracellular cholesterol concentration. To investigate intracellular consequences of persistent LDL receptor overexpression we constructed a recombinant adenovirus encoding the human LDL receptor under control of the Rous sarcoma virus promoter. The metabolic and morphological effects of LDL receptor expression were characterized by uptake experiments with human hepatoma cells using fluorescent and radiolabeled LDL. We observed that large amounts of LDL accumulate within LDL receptor transduced cells, which eventually lead to massive intracellular lipid deposition. Kinetic experiments with LDL-supplemented medium resulted in numerous crystal shaped structures in the cytosol of transduced cells as visualized by digital interference contrast optic within 60 min after LDL supplementation. Thin layer chromatography analyses of cellular lipids suggested these crystalline structures to be dependent on intracellular cholesterol and cholesterol ester levels. Mock-infected cells showed neither cholesterol lipid accumulation nor crystal formation. In conclusion, our data demonstrate that nonphysiological overexpression of the LDL receptor can cause massive lipid accumulation, which cannot be compensated by the hepatoma cell metabolism. This phenomenon may result in negative selection of LDL receptor overexpressing cells in vitro and in vivo.

Adenoviruses, Human↗
PubMed1999

Induction of apoptosis and G2/M arrest by infection with replication-deficient adenovirus at high multiplicity of infection.

Replication-deficient adenoviruses are among the most widely used vectors in gene therapy and are also becoming increasingly popular as analytical tools in basic research. However, significant toxicity of these vectors in vivo has been reported. Here, we show that in an in vitro setting, first generation adenoviruses lead to growth retardation, prolongation of the G2/M phase and induction of apoptosis if applied at a high multiplicity of infection (MOI). These findings were obtained in p53-deficient hepatocytes, derived from knock-out mice (A2 cells) and in several tumor cell lines containing wild-type (wt) or mutant p53. Apoptosis induction was correlated with increased levels of p53 and bax proteins and it was stronger in cells containing wt p53 as compared with cells lacking functional p53. Apoptosis was highly dependent on the MOI used with marked effects starting at an MOI twice as high as needed for 100% gene transfer. Expression of the adenoviral E4 ORF6 gene as well as adenoviral replication were detected in all cell lines infected with first generation adenovirus. Apoptosis could be considerably reduced but not abrogated by UV inactivation of adenovirus, which indicates proapoptotic effects caused by the infection event as well as by residual adenoviral gene expression or adenoviral replication. First generation adenoviruses apparently display proapoptotic activity if used at higher MOIs, which may be of relevance when these vectors are used as analytical or gene therapeutic tools.

Adenoviridae↗
PubMed1999

Continuous infusion of FVIII and FIX concentrates: in vitro analysis of clinically relevant parameters.

A high purity factor VIII/von Willebrand Factor (FVIII/vWF) concentrate (IMMUNATE [STIM plus]) (n = 6 batches), and a high purity factor IX (FIX) concentrate (IMMUNINE [STIM plus]) (n = 7 batches), were assessed in vitro for their applicability to continuous infusion. Parameters pertinent to continuous infusion were investigated and included stability, sterility and, in the case of FIX, the generation of potentially thrombogenic components. Four stationary or transportable mini infusion pumps, equipped with polyethylene, polypropylene or polyvinylchloride plastic components were used. The concentrates were reconstituted without extra filling volume and perfused at 12.5 mL h-1 and 1 mL h-1; sampling was carried out at the start of the experiment and for up to 48 h. The FVIII procoagulant activity (FVIII:C) was assayed by amidolytic, 1-stage and 2-stage assays; vWF was examined for ristocetin cofactor activity, antigen and multimers. The FIX coagulation activity (FIX:C) was determined by a 1-stage coagulation assay; thrombogenicity potential was assessed in vivo (Wessler stasis model in rabbits) and in vitro (FIXa and nonactivated thromboplastin time). Reconstituted concentrate incubated under the same conditions served as a control. Both concentrates remained sterile throughout the testing period. The perfused and control samples remained stable, retaining over 95% of activity for FVIII:C and over 90% for FIX:C for up to 48 h. Intermittent decrease of FVIII:C or FIX:C was not observed, suggesting no adsorption of FVIII or FIX onto plastic surfaces during either short or long-term exposure. No thrombogenic components were detected in the high purity FIX concentrate. Thus, under the in vitro conditions used, FVIII/vWF and FIX were found to be suitable for administration by continuous infusion.

Blood Coagulation Tests↗
PubMed1999

Otolaryngology care unit: a safe and cost-reducing way to deliver quality care.

OBJECTIVES: Patients undergoing treatment for head and neck cancer, obstructive sleep apnea, and potential airway obstruction are often unnecessarily admitted to an intensive care unit (ICU). This study determined the efficacy of an intermediate care unit (OtoCare Unit) for their management. METHODS: A mail survey was conducted of 110 academic institutions' experience with intermediate care units; a retrospective study was performed of our ICU use with analysis of the use of invasive monitoring, length of stay, and cost; and a retrospective study of our first 168 OtoCare Unit patients and their outcomes, complications, and charges was performed. RESULTS: There were 56 responses to 110 survey inquiries. Thirty institutions used some form of intermediate care, while five had a separate otolaryngology unit. Analysis of our 1-year ICU experience showed that of 54 patients who underwent head and neck surgery, 36 patients were admitted to the ICU. Of these 36 admissions, only 9 patients required invasive monitoring and the majority had stable clinical courses. Guidelines were established for an OtoCare Unit: patients use non-ICU beds, mobile noninvasive monitoring units are provided, and a 1:4 nurse-to-patient ratio is used. Phase I included 35 patients who required a mandatory post-anesthesia care unit (PACU) stay of 4 hours. Three minor complications occurred in this group. Phase II included 133 patients who were permitted to enter the OtoCare Unit as soon as they recovered from anesthesia. There were nine minor complications and three major complications in this group. The charge savings compared with ICU usage for such patients was $35,762.00. CONCLUSIONS: An OtoCare Unit is a safe and cost-effective means of caring for this select group of patients.

Cost Control↗
PubMed1999

Ovine adenovirus vectors overcome preexisting humoral immunity against human adenoviruses in vivo.

Recombinant human adenoviruses (hAd) have become widely used as tools to achieve efficient gene transfer. However, successful application of hAd-derived vectors in clinical trials is limited due to immunological and potential safety problems inherent in their human origin. In this study, we describe a recombinant ovine adenovirus (OAV) as an alternative vector for gene transfer in vivo. In contrast to an hAd vector, the OAV vector was not neutralized by human sera. An OAV vector which contained the cDNA of the human alpha1-antitrypsin (hAAT) gene linked to the Rous sarcoma virus promoter was generated and administered systemically to mice. The level and duration of hAAT gene expression was similar to that achieved with an hAd counterpart in both immunocompetent and immunodeficient mice. However, the tissue distribution of the OAV vector differed from that observed for hAd vectors in that the liver was not the dominant target. Significantly, we demonstrated efficient gene transfer with the OAV vector into mice immunized with hAd vectors and vice versa. We also confirm that the immune response to a transgene product can prevent its functional expression following sequential application of a vector. Our results suggest a possible solution to endemic humoral immunity against currently used hAd vectors and should therefore have an impact on the design of improved gene therapy protocols utilizing adenovirus vectors.

Adenoviruses, Human↗
PubMed1999

NF-kappaB function in growth control: regulation of cyclin D1 expression and G0/G1-to-S-phase transition.

Nuclear factor kappa B (NF-kappaB) has been implicated in the regulation of cell proliferation, transformation, and tumor development. We provide evidence for a direct link between NF-kappaB activity and cell cycle regulation. NF-kappaB was found to stimulate transcription of cyclin D1, a key regulator of G1 checkpoint control. Two NF-kappaB binding sites in the human cyclin D1 promoter conferred activation by NF-kappaB as well as by growth factors. Both levels and kinetics of cyclin D1 expression during G1 phase were controlled by NF-kappaB. Moreover, inhibition of NF-kappaB caused a pronounced reduction of serum-induced cyclin D1-associated kinase activity and resulted in delayed phosphorylation of the retinoblastoma protein. Furthermore, NF-kappaB promotes G1-to-S-phase transition in mouse embryonal fibroblasts and in T47D mammary carcinoma cells. Impaired cell cycle progression of T47D cells expressing an NF-kappaB superrepressor (IkappaBalphaDeltaN) could be rescued by ectopic expression of cyclin D1. Thus, NF-kappaB contributes to cell cycle progression, and one of its targets might be cyclin D1.

3T3 Cells↗
PubMed1999

Protection of baculovirus-vectors against complement-mediated inactivation by recombinant soluble complement receptor type 1.

Baculovirus-based vectors are efficient means for gene transfer into hepatocytes in vitro. However, gene transfer in vivo is hampered by inactivation of baculovirus by the complement system. In this study, we demonstrate protection of baculovirus vectors against complement-mediated inactivation through recombinant soluble complement receptor type 1 (sCR1). Blocking of only the alternative complement pathway by a mutant of sCR1 did not result in baculovirus survival in human serum. The data suggest the use of sCR1 as a potent drug to facilitate baculovirus-mediated gene transfer into hepatocytes in vivo.

Animals↗
PubMed1999

Pancreas ultrastructural alterations in mice inoculated with Tityus discrepans (Buthidae) venom.

The symptoms of scorpionic envenomation in mice appear almost immediately after intraperitoneal injection and are manifested by great agitation, hair bristling, accelerated respiration, salivation and lacrimation, vomits and diarrhoea. In this work we intend to correlate those clinical manifestations appearing in response to the toxic aggression by Tityus discrepans venom, to the cellular or subcellular alterations produced in the mouse pancreas, probably similar to those damages found in envenomed humans. To evaluate pancreas subcellular response to Tityus discrepans venom, male C57/Bl adult mice were randomised into two groups: envenomed were intraperitoneally injected (hypochondrial left region) at a dose of 5 mg/Kg weight and controls received saline solution. Samples after preparation were studied in a Hitachi-300 transmission electron microscope. The most relevant ultrastructural changes in pancreatic tissues were an increase in the nuclear heterochromatin, with a corresponding decrease of euchromatin. In the cytoplasm, rough endoplasmic reticulum exhibited zones of oedema, losing its organised aspect. The secretion granules presented smaller electron density and variability in dimensions. At higher magnification a nucleus with picnotic appearance, with indentation of its perinuclear cistern was observed. There was a mitochondrial degeneration, with destruction of the mitochondrial matrix and autophagic vacuoles in its interior. At 48 h the lesions became intensified, with an evident increase in the intercellular spaces.

Animals↗
PubMed1998

p21WAF1/CIP1 mutants deficient in inhibiting cyclin-dependent kinases (CDKs) can promote assembly of active cyclin D/CDK4(6) complexes in human tumor cells.

The cyclin-dependent kinase (CDK) inhibitor p21WAF1/CIP1 is a multidomain, multifunctional protein and a candidate tumor suppressor. Here, we show that, among rationally designed and tumor-associated mutants of human p21 ectopically expressed in U-2-OS cells, those that are selectively deficient in binding to either cyclin or CDK are partially impaired in inhibiting endogenous CDK activities but efficiently promote assembly of active cyclin D/CDK4(6) complexes. These results provide mechanistic insights into the p21-cyclin/CDK interplay in vivo and suggest a functional subclassification of tumor-specific aberrations of p21. Intriguingly, the subclass exemplified by the melanoma-derived N50S mutant may promote tumorigenesis, by both attenuating CDK-inhibitory function and concomitantly activating the proto-oncogenic cyclin D-dependent kinases.

Cyclin D↗
PubMed1998

Expression of a p16INK4a-specific ribozyme downmodulates p16INK4a abundance and accelerates cell proliferation.

The pl6INK4a tumor suppressor negatively regulates progression through the G1 phase of the mammalian cell cycle. To mimic the downmodulation of p16INK4a commonly seen in cancer, we designed and characterized a hammerhead ribozyme against exon E1alpha of the murine pl6INK4a transcript. Stable expression of the ribozyme in murine erythroleukemia (MEL) cells reduced the endogenous pl6INK4a protein by more than 70% and significantly accelerated cell cycle progression. The specificity and efficiency of our new ribozyme suggest its possible application in elucidating the role of p16INK4a in fundamental biological processes including homeostatic tissue renewal, protection against oncogenic transformation, and cellular senescence.

Animals↗
PubMed1998

Cyclin D3: requirement for G1/S transition and high abundance in quiescent tissues suggest a dual role in proliferation and differentiation.

The mammalian D-type cyclins D1, D2, and D3 activate the cyclin-dependent kinases CDK4 and CDK6 in G1 and thereby promote the cell's commitment to enter S phase. To elucidate the extent of functional overlap among the D-type cyclins, we have examined several aspects of the least characterized member of this subfamily of G cyclin proteins, cyclin D3. Microinjection of cyclin D3-neutralizing antibody inhibited G1/S transition in human (IMR-90) and rat (R12) diploid fibroblasts, indicating that analogous to cyclins D1 and D2, cyclin D3 is essential for timely progression through G1. In contrast to cyclins D1 and D2, cyclin D3 was (i) ubiquitously expressed among a panel of 70 human cultured cell types; (ii) strongly upregulated upon induction of HL-60 leukaemia cells to differentiate; and (iii) accumulated to high levels in a wide range of quiescent cell types in mouse and human differentiated tissues. Complementary analyses of human biopsies and mouse tissues at different stages of foetal and postnatal development revealed lineage-dependent transient or long-term accumulation of the cyclin D3 protein, correlating with initiation/establishment or maintenance of the mature phenotypes, respectively. Our data support the notion that the biological roles of the individual D-type cyclins are not fully redundant, and suggest a possible dual role for cyclin D3 in cell proliferation and induction and/or maintenance of terminal differentiation.

Antibodies, Monoclonal↗
PubMed1998

Carnitine promotes heat shock protein synthesis in adriamycin-induced cardiomyopathy in a neonatal rat experimental model.

In order to evaluate carnitine protective strategy and its relationship with heat shock protein induction, female Sprague-Dawley neonatal rats, body weight 40 g, were randomized into four groups: control, adriamycin, carnitine and carnitine-adriamycin. Adriamycin was injected i.v. at a dose of 27 mg/kg (0.1 ml). Carnitine was administered i.v. (20 mg/0.1 ml) before each subdose of adriamycin and then per os (180 mg/kg) daily for 12 weeks. Body weight was recorded weekly. Ventricular wall thickness and cellular damage percentage were morphometrically and ultrastructurally determined, respectively. The determinations were realized monthly until the third month after treatment. The heat shock protein 25 content in the supernatant of the homogenized heart tissue was determined by Western blot analysis. Eight and 12 weeks after treatment, body weight and ventricular wall thickness decreased much more in adriamycin groups than in control and carnitine ones. At the same time, electron microscopic analysis of adriamycin left ventricular wall samples showed loss of myofibrils, swollen mitochondria and vacuoles. Carnitine-adriamycin treated rats resemble control groups more than adriamycin treated samples. Moreover, de-novo synthesis of heat shock protein was three times more induced in carnitine-adriamycin rats than in adriamycin ones. Carnitine may enhance the cell-protecting mechanism based on an induction of shock protein, and this first cellular response could reduce the severity of late adriamycin-cardiomiopathy.

Animals↗
PubMed1998

Biocompatibility and blood-aqueous barrier impairment in at-risk eyes with heparin-surface-modified or unmodified lenses.

PURPOSE: To evaluated the influence of heparin-surface-modified (HSM) versus unmodified poly(methyl methacrylate) (PMMA) intraocular lenses (IOLs) on the blood-aqueous barrier (BAB) in at-risk eyes. SETTING: Department of Ophthalmology, Bundesknappschaft's Hospital, Sulzbach Germany. METHODS: This study comprised 100 patients with predisposing risk factors for BAB destabilization (e.g., diabetes mellitus with or without retinopathy, glaucoma, pseudoexfoliation, uveitis). One eye in each patient received an HSM IOL and the fellow eye, a conventional unmodified PMMA IOL after phacoemulsification by the same surgeon. Anterior chamber flare was measured with the Kowa 500 laser flare meter 1 day before and 1 day, 1 and 6 weeks, and 3 months after surgery. RESULTS: For most risk factors, mean flare was lower in the HSM group than in the PMMA group at most follow-ups. Significantly lower flare values (difference between postoperative and preoperative mean values) were seen in eyes with the HSM IOL at 6 weeks (P < .004) and 3 months (P < .003; Student's t-test). In the group with preoperative elevated flare values, the eyes with the HSM IOL had significantly better results 6 weeks (P < .0006) and 3 months (P < .01) postoperatively. The values in the HSM IOL eyes were also significantly higher in the diabetic with retinopathy group at 3 months (P < .003). CONCLUSION: The results confirm the efficacy of IOL surface modification in reducing postoperative intraocular reaction in at-risk eyes.

Biocompatible Materials↗
PubMed1998

Transient immunosuppression with 15-deoxyspergualin prolongs reporter gene expression and reduces humoral immune response after adenoviral gene transfer.

A strong immune response against transgenic cells is one important limitation for long-term expression after adenoviral gene transfer in mammals. Continuous pharmacological immunosuppression has been shown to ameliorate immune reactions and to prolong reporter gene expression. In this study, we explored the effect of short-term immunosuppression for long-term gene expression and its impact on antibody formation. Immunosuppression with FK 506 (1 mg/kg/day), cyclosporin A (20 mg/kg/day) and 15-deoxyspergualin (10 mg/kg/day) was performed in NMRI mice. Expression of the reporter gene human alpha-1-antitrypsin (hAAT) and antibody formation was monitored for 7 months. A 5-day course of 15-deoxyspergualin (15-DSG) markedly slowed the decline of reporter gene expression and a positive effect was still detectable 200 days after gene transfer. At the same time, antibody production was reduced by 50-60%. Continuous treatment with 15-DSG (10 mg/kg twice weekly) led to a further small increase of gene expression but reduced antibody formation by 80-90%. A short course of FK 506 and cyclosporin A (CsA), had conferred a negative effect on gene expression. Both groups showed an even faster reduction in gene expression compared with the control group. The results of this investigation suggest that 15-DSG could serve as an effective supplement for viral gene therapy protocols.

Adenoviridae↗
PubMed1998

Baculovirus-mediated gene transfer in the presence of human serum or blood facilitated by inhibition of the complement system.

Baculovirus vectors are efficient tools for gene transfer into hepatocytes in vitro. However, gene transfer is strongly reduced in the presence of native sera, providing an explanation for the failure of direct application of the virus in vivo. In this study, we define the role of the complement (C) system (C) as a major cause for baculovirus inactivation in human serum. Baculoviruses most likely activate the classical pathway of the C system and assembly of very late C components is required for inactivation of the vector. We demonstrate the survival of baculovirus vectors in human serum through treatment with a functional blocking antibody against C component 5. Inactivation of baculovirus in human plasma and whole blood was prevented by treatment with cobra venom factor. The data reveal various interactions of baculovirus vectors with the C system and will lead to facilitation of baculovirus-mediated gene transfer into hepatocytes in vivo by protection of the vector from C inactivation.

Antibodies, Monoclonal↗