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Biomedical subjects

M Pagano

Publications and source records attributed to M Pagano.

At least 109 records · Page 6Linked to original sources
PubMed1997

Cell cycle regulation by the ubiquitin pathway.

In the past 2 years, two ubiquitin-dependent proteolytic pathways have been established as important players in the regulation of the cell division cycle. In S. cerevisiae, the entry into S phase requires ubiquitin-mediated degradation of a cdk inhibitor, p40Sic1, in a pathway that involves the E2 enzyme Cdc34. Recent studies reviewed herein show that the Cdc34 pathway targets phosphorylated substrates. A second pathway that regulates chromosome segregation and mitotic exit by degrading anaphase inhibitors and mitotic cyclins involves a different E2 and a large molecular weight E3 complex, called the anaphase-promoting complex or cyclosome. This pathway targets substrates containing one or more destruction box motif.

Anaphase-Promoting Complex-Cyclosome↗
PubMed1997

Kip1 degradation via the ubiquitin-proteasome pathway.

The cell cycle has been the object of extensive studies for the past years. A complex network of molecular interactions has been identified. In particular, a class of cell cycle inhibitory proteins has been identified but details of the molecular mechanism of their action have yet to be resolved. These inhibitors regulate the progression through G1 and the G1/S transition via the inhibition of the cyclin-dependent kinase (Cdk) activity. The potential function of these negative regulators as tumor suppressors provides new insights into the link between the cell cycle and oncogenesis. Kip1 is a potent inhibitor of Cdks. In quiescent cells Kip1 accumulates without an increase in mRNA or protein synthesis. We demonstrated that cell cycle regulation of Kip1 levels, both in normal and transformed human cells, occurs via the ubiquitin-proteasome pathway. In a crude in vitro system, Kip1 is ubiquitinated and degraded in an ATP dependent manner and inhibition or depletion of the proteasome blocks Kip1 degradation. Human Ubc2 and Ubc3, the homologs of yeast Rad6 and Cdc34 gene products respectively, are specifically involved in the ubiquitination of Kip1. Compared to proliferating cells, quiescent cells contain a far lower amount of Kip1 ubiquitinating activity. These results represent the first demonstration that the ubiquitin-proteasome pathway plays a role in the regulation of a cell cycle protein in human cells, namely the Cdk inhibitor Kip1. The specific proteolysis of Kip1 may be involved in the pathway of inactivation of Cdks.

Adenosine Triphosphate↗
PubMed1996

The association between youth, women, and acquired immunodeficiency syndrome.

This article compares the characteristics of women and heterosexual men with AIDS in New York City. The analysis was performed using the New York City AIDS Surveillance Database, namely, those 37,002 persons diagnosed from 1984 to 1993 between ages 15 and 64, excluding men who report sex with other men as their sole risk behavior. The median age at diagnosis was 34 years for women with heterosexually acquired disease, 36 years for women with a history of injection drug use, and 39 years for men, most of whom used injection drugs (p < 0.001). The proportion of women and the rate of increase of this proportion were greater among younger AIDS cases. By 1993 women comprised the majority of cases under age 30, and most of these young women had heterosexually acquired disease. For each decrease in 5-year age group under age 45, the odds of a case being a woman increased by 30% (95% confidence interval = 27, 33%) after adjustment for year. CD4 cell count reporting, and race/ethnicity. There was a somewhat greater youth-gender effect among black persons with AIDS (6% additional increase for each decrease in age group; 95% confidence interval = 3, 10%). Therefore, women are overrepresented among younger persons with AIDS, particularly persons of color. They are largely infected through heterosexual contact with men who have used intravenous drugs.

Acquired Immunodeficiency Syndrome↗
PubMed1996

Competition between plasminogen and procathepsin B as a probe to demonstrate the in vitro activation of procathepsin B by the tissue plasminogen activator.

The tissue plasminogen activator (tPA) was found to activate in vitro the procathepsin B purified from malignant ascitic fluids. This activation was time and dose dependent, and was associated with the processing of procathepsin B. The present study shows that tPA is a fast activator of procathepsin B in a neutral pH range, such that generation of cathepsin B activity and processing of procathepsin B are achieved after a 5-min incubation time at 37 degrees C, pH 7.4. In contrast, competition between plasminogen and procathepsin B was observed for the activation and processing by tPA. From these findings, a plasminogen activator pathway for procathepsin B activation related to the plasminogen concentration may exist. In vivo this pathway may be involved in a proteolytic cascade linked to invasion and metastasis.

Binding, Competitive↗
PubMed1996

Immunopurification and characterization of a collagenase/gelatinase domain issued from basement membrane fibronectin.

The proteolytic potential of cellular fibronectin fragments issued from a basement membrane hydrolysate was investigated. Three different gelatinase activities (47, 43 and 37 kDa), located by gelatin zymography, were isolated using successively heparin-agarose, gelatin-agarose and immunopurification with polyclonal antibodies directed against bovine plasma fibronectin. These fragments were also characterized using a monoclonal antibody directed against the extra-domain EDA of cellular fibronectin as a probe. A collagenase activity, reliably indicated by the gelatin zymography pattern, was also found using MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2, the intramolecularly quenched fluorogenic substrate of collagenases. From these results, cellular fibronectin was found to be able to exhibit a proteolytic function after limited proteolysis. This MMP-like function could be associated with tissue remodeling in both normal and pathological states, such as metastasis, angiogenesis and tissue repair.

Amino Acid Sequence↗
PubMed1996

A 39 amino acid fragment of the cell cycle regulator p21 is sufficient to bind PCNA and partially inhibit DNA replication in vivo.

The cell cycle regulator p21 interacts with and inhibits the DNA replication and repair factor proliferating cell nuclear antigen (PCNA). We have defined a 39 amino acid fragment of p21 which is sufficient to bind PCNA with high affinity (Kd 10-20 nM). This peptide can inhibit DNA replication in vitro and microinjection of a GST fusion protein containing this domain inhibited S phase in vivo. Despite its high affinity for PCNA, the free 39 amino acid peptide does not have a well-defined structure, as judged from circular dichroism and nuclear magnetic resonance measurements, suggesting an induced fit mechanism for the PCNA-p21 interaction. The association of the small peptide with PCNA was thermolabile, suggesting that portions of p21 adjoining the minimal region of contact stabilize the interaction. In addition, a domain containing 67 amino acids from the N-terminus of PCNA was defined as both necessary and sufficient for binding to p21.

Amino Acid Sequence↗
PubMed1996

Cell cycle regulation of nuclear factor p32 DNA-binding activity by novel phase-specific inhibitors.

The nuclear factor p92, originally discovered by its interaction with the human papillomavirus type 18 enhancer, is a cellular protein whose activity is restricted to S phase in human primary fibroblasts. The human papillomavirus type 18 p92 binding sequence confers enhancer activity on a heterologous promoter, suggesting that p92 acts as a transcription factor. We have identified a class of nuclear inhibitory proteins, I-92s, which noncovalently associate with p92 but not with other transcription factors such as AP1, E2F, or NF-kappaB. Different I-92s occur in G1, G2, and G0, while no I-92 is detectable in S phase. Phase-specific inhibitors, therefore, are responsible for the cell cycle dependence of p92 activity and provide a novel mechanism linking transcription factor regulation with the cell cycle.

Base Sequence↗
PubMed1996

A 67 kDa non-hormone binding estradiol receptor is present in human mammary cancers.

The presence of large amounts of a 67 kDa estradiol receptor that does not bind hormone was observed in 8 to 37 human mammary tumors (34 malignant and 3 benign). This form of receptor was detected by its conversion to hormone binding receptor by an endogenous tyrosine kinase in vitro. All 8 tumors were malignant. In these, the incubation of cytosol with ATP was seen to cause a 1- to 5-fold increase in estradiol-specific binding sites. These sites bound estradiol with physiological affinity, and their appearance was associated with tyrosine phosphorylation of estradiol receptor. The enzyme converting the non-hormone binding receptor into the hormone binding receptor is largely present in cytosol and scarce in membranes. It has been extensively purified. It is a 67 kDa protein under denaturating conditions, binds calmodulin-Sepharose in a Ca2+-dependent manner, is stimulated by Ca2+ and calmodulin, phosphorylates exogenous actin, is activated by the estradiol-receptor complex. The enzyme interacts with antibodies directed against the carboxy-terminal and catalytic domains of c-src. Therefore, it is a putative new member of the large c-src-related kinase family. Human mammary cancers with significant amounts of 67 kDa non-hormone binding receptor show relatively low levels of hormone binding estradiol receptor. The presence of non-hormone binding receptor that can be activated by in vitro tyrosine phosphorylation suggests that functional interaction of estradiol receptor with tyrosine kinases is altered in malignant tumors and has bearing on loss of hormone dependence and progression of the mammary cancer malignancy.

Adenosine Triphosphate↗
PubMed1996

Life tables.

Explore the source record for details and available documents.

Adolescent↗
PubMed1996

Screening for psychosocial problems in 4-5-year-olds during routine EPSDT examinations: validity and reliability in a Mexican-American sample.

The effectiveness of the Pediatric Symptom Checklist (PSC) as a psychosocial screening measure to meet Federal Medicaid/Early and Periodic Screening, Diagnosis, and Treatment (EPSDT) requirements was examined in 117 low-income preschool (aged 4-5 years old) Hispanic children during well-child examinations in three clinics over an 8-month period. The PSC identified 7% of the sample as at risk for psychosocial problems. The PSC was significantly associated with parental ratings of the children's problems in functioning, with pediatric clinicians' decisions to make mental health referrals, with degrees of associations similar to those found between PSC scores, and with the same measures with school-aged children in the same clinics. Cronbach's alpha was high (r = .87) and virtually identical in English, Spanish, oral, and written formats. Although it identified a slightly lower rate of psychosocial problems in 4-5-year-olds than it had in school-aged children, the PSC appeared to provide an effective method of screening for psychosocial problems during EPSDT examinations.

California↗
PubMed1996

Cell cycle and cancer: critical events at the G1 restriction point.

In eukaryotic cells, each phase of the cell division cycle is controlled by the sequential activation of various cyclin-dependent kinases (Cdks). These kinases are known to phosphorylate various substrates whose activity is critical for cell cycle progression. As key regulators of the cell cycle, Cdks must be strictly controlled by both extracellular and intracellular signals for adequate responses to occur. There are several distinct molecular mechanisms for controlling the activity of the different Cdks: regulated synthesis and destruction of the activating subunit (cyclin), regulated synthesis and destruction of the inhibitory subunit (Cki), and posttranslational modification of the kinase subunit by highly specific kinases and phosphatases. During the G1 phase of the cell cycle, cells sense, integrate positive and negative signals, and transmit them to the cell cycle machinery. Because of this pivotal role, a vast majority of oncogenic events selectively target elements controlling the G1. In this review we discuss the elements controlling the G1 phase in relationship to the genesis of cancer.

Cyclin-Dependent Kinases↗
PubMed1996

Persistent pulmonary hypertension of the newborn and smoking and aspirin and nonsteroidal antiinflammatory drug consumption during pregnancy.

OBJECTIVE: Prenatal causation of persistent pulmonary hypertension of the newborn (PPHB) is suggested by a specific pattern of pulmonary vascular remodeling observed immediately after birth in some infants with fatal PPHN. The goal of this study was to determine whether PPHN is associated with fetal exposure to: (1) tobacco and marijuana smoking (ie, contributors to fetal hypoxemia), (2) consumption of aspirin and other nonsteroidal antiinflammatory drugs (ie, inhibitors of prostaglandin synthesis), and (3) cocaine use (ie, a contributor to vasospasm). DESIGN: Case-control interview study. SETTING: Two Harvard-affiliated newborn intensive care units. PARTICIPANTS: Mothers of case infants who had PPHN or who met criteria for the referent group. INTERVENTIONS: During July 1985 through April 1989, we interviewed mothers of 103 infants with PPHN and 298 control infants. Because of potential selection bias that might result from recruiting only inborn control infants even though two-thirds of cases were outborn, separate analyses compared the 103 total and 35 inborn infants with PPHN with the 298 inborn control infants. Multivariate analyses were used to adjust for potential confounding factors, including maternal education and Medicaid health insurance (ie, two markers of socioeconomic status), other antenatal factors found to be associated with PPHN (ie, maternal urinary tract infection and diabetes mellitus), and the infant's sex. MAIN OUTCOME MEASURES: Self-reported use or consumption of tobacco, marijuana, cocaine, aspirin, and other nonsteroidal antiinflammatory drugs during pregnancy. RESULTS: The adjusted odds ratios (and 95% confidence intervals) for maternal pregnancy exposures to the factors of principal interest among the total study population were: aspirin, 4.9 (1.6-15.3); and nonsteroidal antiinflammatory drugs, 6.2 (1.8-21.8); for the inborn group they were aspirin, 9.6 (2.4-39.0); and nonsteroidal antiinflammatory drugs, 17.5 (4.3-71.6). Although the association between tobacco smoking during pregnancy and PPHN was elevated in univariate analyses, with odds ratios (and 95% confidence intervals) of 2.0 (1.2-3.4) and 1.3 (0.6-3.3) for total and inborn populations, respectively, the relationship was not significant after adjustment for all other factors in the final logistic regression model. Acknowledged illicit drug use was too infrequent (3.2%) to evaluate. CONCLUSION: Maternal consumption of nonsteroidal antiinflammatory drugs and aspirin during pregnancy or the reasons these drugs were ingested seem to contribute to an increased risk of PPHN.

Adult↗
PubMed1996

[Wegener granulomatosis: a clinical case with parossistic positional vertigo due to involvement of the lateral semicircular canal].

The ear is involved in more than 20% of all cases of Wegener Granulomatosis (W.G.): such involvement takes place either through direct proliferation of the necrotic-granulomatosis tissue in the middle ear, or is a consequence of perieustachian tube infiltration from the rhinopharnyx. The inner ear is affected later and it is not clear as to exactly how. The present paper deals with the case of a 63-year-old woman who had a year been suffering from bilateral ear pain, and persistent otorrhea with typical signs of purulent chronic middle-ear otitis. During the course of the illness the patient complained of atypical paroxymal positional vertigo, compatible with cupulolithiasis of the semi-circular horizontal canal (as per E.N.G. recording). Transoral biopsy of the rhinopharynx and of the left nasal fossa revealed histopathological signs compatible with W.G.. In the light of post-mortem temporal bone studies performed by Friemann and Blatt on W.G. patients the present case could lead to further studies regarding the advancement of this pathological process within the middle and inner ears and the etiopathogenetic mechanisms involved. In particular, during the course of W.G., the appearance of benign paroxymal positional vertigo through cupulolithiasis of the semicircular horizontal canal may be justified: a) by the progressive involvement of the posterior labyrinth structures affecting the semicircular canals prior to the lateral canal and running on to the posterior canal; b) by the discovery of proteiform material in the labyrinthine fluids.

Cerebrospinal Fluid Otorrhea↗
PubMed1995

Identification of a Myc-dependent step during the formation of active G1 cyclin-cdk complexes.

Activation of conditional alleles of Myc can induce proliferation in quiescent cells. We now report that induction of Myc in density-arrested fibroblasts triggers rapid hyperphosphorylation of the retinoblastoma protein and activation of both cyclin D1- and cyclin E-associated kinase activities in the absence of significant changes in the amounts of cyclin-cdk complexes. Kinase activation by Myc is blocked by inhibitors of transcription and requires intact DNA binding and heterodimerization domains of Myc. Activation of cyclin E-cdk2 kinase in serum-starved cells occurs in two steps. The first is induced by Myc and involves the release of a 120 kDa cyclin E-cdk2 complex from a 250 kDa inactive complex that is present in starved cells. This is necessary, but not sufficient, to generate full kinase activity, as cdc25 phosphatase activity is limiting in the absence of external growth factors. In vivo cdc25 activity can be supplied by the addition of growth factors. In vitro recombinant cdc25a strongly activates the 120 kDa, but only poorly activates the 250 kDa cyclin E-cdk2 complex. Our data show that two distinct signals, one of which is supplied by Myc, are necessary for consecutive steps during growth factor-induced formation of active cyclin E-cdk2 complexes in G(o)-arrested rodent fibroblasts.

Animals↗
PubMed1995

Inhibition of chondrocyte cathepsin B and L activities by insulin-like growth factor-II (IGF-II) and its Ser29 variant in vitro: possible role of the mannose 6-phosphate/IGF-II receptor.

Lysosomal enzymes and IGF-II both bind to the mannose 6-phosphate (M6P)/IGF-II receptor. This receptor targets newly synthesized lysosomal enzymes to lysosomes. The functional meaning of IGF-II binding to this receptor is not well known. We have postulated that IGF-II, the Ser29 IGF-II variant (vIGF-II) and IGF-I on lysosomal cathepsin B and L activities from post-natal rabbit chondrocytes in vitro. This effect was compared with the ability of each peptide to stimulate chondrocyte-sulfated proteoglycan synthesis. The sulfating dose-response relationship of the IGF peptides corresponded to their relative binding affinities for the type I-IGF receptor (IGF-I > IGF-II > vIGF-II). The intracellular cathepsin B and L activities were inhibited in a time- and dose-dependent manner by IGF-II or vIGF-II. Maximal inhibition of cathepsin B and L activities (40 and 30% below controls, respectively) was found after an 8 h treatment with 100 ng/ml IGF-II or vIGF-II. By contrast, IGF-I up to 1 micrograms/ml or insulin up to 2 micrograms/ml had no inhibitory effect. The relative potency pattern corresponded to the binding profile of each ligand for the M6P/IGF-II receptor. A treatment of chondrocytes with IGF-I or insulin transiently increased the binding of radiolabelled IGF-II at the cell surface to approximately 120% of controls, whereas IGF-II or vIGF-II had no effect. Thus, it is unlikely that the inhibition of lysosomal enzyme activities by IGF-II peptides could result from a redistribution of M6P/IGF-II receptors from intracellular compartments to the plasma membrane. We hypothesize that internalized IGF-II peptides could occupy the intracellular M6P/IGF-II binding sites required for targeting of cathepsins B and L to lysosomes.

Animals↗