Biomedical subjects

R A Perugini

Publications and source records attributed to R A Perugini.

14 recordsLinked to original sources

PubMed2004

Hand-assisted laparoscopic splenectomy in the setting of splenomegaly.

BACKGROUND: Hand-assisted laparoscopic surgery (HALS) devices may be well suited to splenectomy in cases of splenomegaly. METHODS: All cases of HALS for splenectomy between 1997 and 2001 were reviewed. Patient characteristics, operative details, and morbidity and mortality were analyzed. RESULTS: HALS for splenectomy was performed in 54 patients. A total of 39 patients with massive splenomegaly (MS) (>600 g) were identified. The average weight of the MS group was 1285 +/- 505 g. There was one (3%) conversion. Operative time was 159 +/- 65 min, estimated blood loss was 257 +/- 240 ml, and length of hospital stay was 5.4 +/- 2.9 days. Morbidity was limited to 13 patients (24%), and there were two postoperative mortalities (5.1%). CONCLUSIONS: HALS for splenectomy in the setting of splenomegaly is feasible and safe. For the surgeon considering a laparoscopic approach in the setting of splenomegaly, a hand-assisted technique is ideally suited for removal of the enlarged spleen.

Adult↗

PubMed2004

Use of a laparoscopic hand-assist device for accessory splenectomy.

BACKGROUND: The use of a laparoscopic hand-assist device may aid in the identification of accessory spleens (ASs) and provide similar benefits to a conventional laparoscopic procedure. A patient with previous splenectomy for immune thrombocytopenic pupusa (ITP) and recurrent thrombocytopenia is reported. METHOD: A computed tomography scan and RBC scan identified several nodules consistent with ASs. Initial laparoscopic exploration could not identify all the ASs seen on preoperative imaging. A hand-assist device was placed and a total of five nodules of splenic tissue were identified without conversion to laparotomy. RESULTS: The patient had a brief and uncomplicated postoperative course with a return of platelet counts to 350,000 at 1-month follow-up. CONCLUSION: We propose that in the scenario of recurrent ITP following laparoscopic splenectomy, repeat laparoscopy is the first step once an AS is identified by preoperative imaging. If the AS is not identified at laparoscopy, the insertion of a hand-assist device is an alternative to a full laparotomy.

Congenital Abnormalities↗

PubMed2001

FRAP-p70s6K signaling is required for pancreatic cancer cell proliferation.

BACKGROUND: FRAP-p70s6K signaling regulates mitogenic responses to growth factors in eukaryotic cells. Constitutive p70s6K activation occurs in some human malignancies and may contribute to dysregulated cell growth. We examined whether inhibition of this pathway affects mitogen-induced proliferation and cell cycle progression of human pancreatic cancer cells in vitro. METHODS: Quiescent BxPC3 and Panc-1 human pancreatic cancer cells treated with or without 20 ng/mL rapamycin (FRAP inhibitor) were repleted with 10% FCS to induce cell cycle entry. Proliferation was measured with MTT assay. Cell cycle and apoptosis were determined by FACS analysis. Phosphorylation of p70s6K, Akt, and cdc2 was evaluated by Western blot. Statistical analysis was by two-tailed t test (P < 0.05). RESULTS: Rapamycin (Rapa) inhibited the phosphorylation of p70s6K while inducing G(1) cell cycle arrest (P < 0.005). In both cell lines, Rapa inhibited serum-induced proliferation (P < 0.05) without affecting apoptosis. Cdc2 phosphorylation was inhibited by 15 min with Rapa (not shown), consistent with cell cycle arrest. Akt phosphorylation was not affected, indicating FRAP specificity of Rapa. CONCLUSIONS: FRAP-p70s6K signaling appears to be necessary for G(1)-to-S phase progression and proliferation in pancreatic cancer cells. This supports earlier work demonstrating a similar regulatory role for PI-3' kinase, an upstream activator of FRAP-p70s6K.

Adenocarcinoma↗

PubMed2000

Pancreatic cancer cell proliferation is phosphatidylinositol 3-kinase dependent.

BACKGROUND: Genetic mutations found in pancreatic cancer (K-ras, p16, p53) lead to inappropriate cellular proliferation. Mitogens stimulate proliferation via the phosphatidylinositol 3-kinase (PI3K)- and/or the p44/42-mitogen-activated protein kinase [p44/42-MAPK or extracellular signal-regulated kinase (ERK)] signaling pathways. We examined whether inhibition of either PI3K or ERK could limit proliferation in human pancreatic cancer. METHODS: Proliferation was stimulated in quiescent human pancreatic cancer cell lines (BxPC3 and Panc-1) by 10% fetal calf serum (FCS). In certain samples, PD98059 (an ERK inhibitor) or LY294002 (a PI3K inhibitor) was also added. AKT phosphorylation (indicating PI3K activity) and ERK phosphorylation (ERK activation) were determined by Western blot. Cell viability was determined by MTT assay. Cell cycle progression and apoptosis were determined by flow cytometry. A two-tailed t test was used for statistical analysis of the data (significance P < 0.05). RESULTS: LY294002 inhibited the PI3K pathway without affecting ERK activation in response to serum. PD98059 inhibited the ERK pathway specifically. In both BxPC-3 and Panc-1 cell lines, LY294002 inhibited serum-induced proliferation. This was associated with G(1) cell cycle arrest and with an increase in the rate of apoptosis. PD98059 inhibited proliferation only in BxPC3 cells, and to a lesser degree than did LY294002. CONCLUSIONS: PI3K signaling appears to be necessary for G(1)-to-S phase progression and proliferation in pancreatic cancer cells. ERK plays a lesser role in mitogen-induced proliferation. Pharmacological inhibition of PI3K may decrease proliferation, increase apoptosis, and potentially confer therapeutic benefit in pancreatic cancer.

Cell Cycle↗

PubMed2000

NFkappaB expression during cold ischemia correlates with postreperfusion graft function.

In liver transplantation, activation of NFkappaB occurs upon reperfusion, yet few data exist regarding NFkappaB activation during cold ischemia. We hypothesized that activation of NFkappaB may initially occur during cold ischemia, prior to reperfusion, and serve as an important determinant of postreperfusion function. To test this hypothesis, serial biopsies during porcine liver harvest were obtained immediately upon laparotomy, upon completion of dissection, after 45 and 120 min of cold ischemia, and 60 and 180 min after reperfusion. Nuclear extracts were isolated for Western blot analysis of NFkappaB. Hepatic function was assessed through bile output and sorbitol dehydrogenase (SDH) activity. NFkappaB expression was maximal at 45 min of cold ischemia and decreased by 120 min. The expression at 120 min of cold ischemia correlated with markers of postreperfusion function, namely bile flow and SDH activity. During reperfusion a second distinct peak occurred at 180 min. Increased expression of NFkappaB at 180 min of reperfusion correlated directly with prior expression at 120 min during cold ischemia and with increased SDH activity. These data indicate that nuclear expression of NFkappaB demonstrate two distinct peaks of activity, one during cold ischemia and one after reperfusion. Enhanced expression of NFkappaB during cold ischemia not only correlates directly with NFkappaB expression during reperfusion, but also correlates inversely with postreperfusion graft function.

Animals↗

PubMed2000

Laparoscopic surgery and Kupffer cell activation.

BACKGROUND: Evidence tends to support a relative preservation of the systemic immune response with laparoscopy as compared with laparotomy. However, the role of hepatic macrophages, or Kupffer cells, in modulating this immune advantage is unknown. This study investigated the functions of Kupffer cells after either laparoscopy or laparotomy in a rat model. METHODS: Rats underwent laparoscopy, laparotomy, or control operations. Kupffer cells were harvested, cultured, and stimulated with lipopolysaccharide. Culture supernatants were analyzed for tumor necrosis factor (TNF-alpha) and interleukin-6 (IL-6). Cytoplasmic lysates were analyzed for activation of two mitogen-activated protein kinases (MAPKs). RESULTS: Production of TNF-alpha and IL-6 was similar in laparoscopy, laparotomy, and control groups. Both laparotomy and laparoscopy showed increased activation of p38 MAPK as compared with controls. Activation of ERK1/2 was decreased during laparotomy as compared with laparoscopy. CONCLUSIONS: Although cytokine production was similar in the laparoscopy and laparotomy groups, changes in MAPK activation suggest that intracellular pathways are more affected during laparotomy than during laparoscopy.

Animals↗

PubMed1999

Sodium salicylate inhibits macrophage TNF-alpha production and alters MAPK activation.

INTRODUCTION: Transcriptional activation of the TNF-alpha gene in LPS-stimulated macrophages is dependent upon nuclear factor kappa-B (NF-kappaB) activity. Salicylates may interfere with NF-kappaB activity through a MAPK (mitogen-activated protein kinase)-dependent process. These studies investigate the effects of sodium salicylate (NaSal) on TNF-alpha production and MAPK activation in macrophages. METHODS: Rat peritoneal macrophages were pretreated or not with sodium salicylate or ibuprofen for 1 h and then stimulated with 100 ng/ml LPS. Six hours following stimulation, cell viability was assessed by MTT assay. At specified time intervals after LPS stimulation, supernatant TNF-alpha was measured by ELISA. Western blots of cell lysates were performed for analysis of total and activated (phosphorylated) MAPKs. RESULTS: Salicylate and LPS, alone or combined, did not significantly alter macrophage viability. Salicylate, but not ibuprofen, significantly reduced TNF-alpha production in LPS-stimulated macrophages. LPS-stimulated activation of ERK and SAPK/JNK was inhibited by NaSal pretreatment. NaSal treatment of macrophages activated p38 MAPK independent of LPS stimulation. Pretreatment of samples with the specific p38 MAPK inhibitor, SB203580, did not significantly alter TNF-alpha production in either LPS or NaSal and LPS-treated samples. CONCLUSIONS: Salicylates alter MAPK signaling and suppress TNF-alpha production in LPS-stimulated macrophages. Salicylate-induced control of inflammatory mediator production in macrophages may, in part, underlie the clinically significant anti-inflammatory effects of these compounds.

Animals↗

PubMed1999

Salicylates inhibit NF-kappaB activation and enhance TNF-alpha-induced apoptosis in human pancreatic cancer cells.

INTRODUCTION: Tumor necrosis factor (TNF-alpha)-induced apoptosis is limited by its coactivation of nuclear factor kappa B (NF-kappaB)-dependent anti-apoptotic genes. Sodium salicylate (NaSal) inhibits NF-kappaB activation by limiting phosphorylation and degradation of its bound inhibitor protein, IkappaB-alpha. We examined whether NaSal enhances TNF-alpha-induced apoptosis in cultured human pancreatic cancer cell lines. METHODS: Two cultured human pancreatic cancer cell lines were studied. PANC-1 and BxPC-3 cells were serum-starved for 12 h, pretreated or not for 1 h with NaSal (5-20 mM), and then stimulated with recombinant human TNF-alpha (400 units/ml). Western blots of cytoplasmic lysates were performed to demonstrate IkappaB-alpha phosphorylation and degradation. Western blots of nuclear extracts were performed to assess nuclear translocation of NF-kappaB. In separate cultures, apoptosis was measured 4.5 h after TNF-alpha stimulation by both ELISA detection of interhistone DNA fragments and flow cytometry with propidium iodide staining. RESULTS: TNF-alpha induced IkappaB-alpha phosphorylation and degradation, which was inhibited by NaSal in both cell lines. TNF-alpha-induced apoptosis (DNA fragmentation) increased significantly when BxPC-3 cells were pretreated with NaSal. Flow cytometry confirmed this, demonstrating increases in apoptotic cell fractions: 8.5% (untreated), 9.3% (TNF-alpha alone), 14.9% (15 mM NaSal), and 22.9% (NaSal and TNF-alpha). In contrast, no increases in apoptosis were measured in the PANC-1 cell line among the various treatment groups. CONCLUSIONS: NaSal enhances TNF-alpha-induced apoptosis while inhibiting IkappaB-alpha phosphorylation and degradation in BxPC-3 human pancreatic cancer cells.

Adenocarcinoma↗

PubMed1999

Ubiquitin-proteasome inhibition enhances apoptosis of human pancreatic cancer cells.

BACKGROUND: Tumor necrosis factor (TNF-a)-induced apoptosis is limited by coactivation of nuclear factor kappa B (NF-kb)-dependent antiapoptotic genes. Nuclear translocation of NF-kB requires degradation of ubiquitinated phospho-IkB-a by the 26S proteasome. We examined whether inhibition of the ubiquitin-proteasome pathway enhances TNF-a-induced apoptosis in BxPC-3 human pancreatic cancer cells. METHODS: Serum-starved BxPC-3 cells (12 hours) were pretreated or not for 50 minutes with PSI (30 m mol/L), a peptide aldehyde known to inhibit specifically the chymotrypsin-like activity of the 26S proteasome. Cells were subsequently stimulated with recombinant human TNF-a (400 units/mL). Western blots were performed using antibodies to IkB-a and phospho-IkB-a. Level of apoptosis was determined by two methods: enzyme-linked immunosorbent assay detection of interhistone DNA fragments and flow cytometry with propidium iodide staining. RESULTS: TNF-a-induced degradation of IkB-a was inhibited by PSI. Phospho-IkB-a accumulation was observed 20 minutes after TNF-a stimulation. Apoptosis relative to constitutive levels was significantly increased after PSI pretreatment, as measured by DNA fragmentation (P < or = .05 by Student t test). Percent apoptosis by flow cytometry confirmed marked increases in apoptotic cell fractions from 5.9% (untreated) to 6.8% (TNF-a alone), 16.4% (PSI alone), and 18.9% (PSI and TNF-a). CONCLUSIONS: PSI enhances both constitutive and TNF-a-induced apoptosis through inhibition of IkB-a degradation in BxPC-3 human pancreatic cancer cells.

Apoptosis↗

PubMed1999

Endotoxin fails to stimulate inositol triphosphate production in macrophages.

UNLABELLED: Inositol Triphosphate (IP3) production is an early cell signaling event which leads to mobilization of intracellular calcium (Ca++). We examined whether bacterial endotoxin (lipopolysaccharide, LPS) stimulates IP3 production in macrophages pretreated with LPS (tolerant) or not. METHODS: RAW 264.7 macrophages were cultured at 5 x 10(6) cells in RPMI supplemented with 10% FCS. LPS tolerance was induced by pretreating macrophages (Tol) for 19 h with 10 ng/ml of LPS. Non-tolerant (Non-Tol) macrophages received no LPS pretreatment. Macrophages were next washed, repleted with fresh media, and stimulated with 100 ng/ml LPS. Paired cultures were stimulated with 1 microM platelet activating factor (PAF), a known stimulant of IP3 production. Following 1, 10, and 15-min stimulation intervals, IP3 was extracted with trichloroacetic acid and measured by receptor displacement assay. RESULTS: LPS did not stimulate IP3 production in either Non-Tol or Tol macrophages. In contrast, PAF stimulated significant increases in IP3 levels within 1 min in both Non-Tol (9.5 +/- 3.0 pmol/ml) and Tol (9.5 +/- 2.4 pmol/ml) macrophages. Non-Tol IP3 levels returned to baseline by 10 min, while Tol IP3 levels remained significantly elevated (8.2 +/- 1.7 pmol/ml). CONCLUSIONS: Unlike PAF, bacterial LPS fails to stimulate IP3 production in macrophages. Furthermore, IP3 production could not be elicited in cultured macrophages repetitively stimulated with LPS.

Cell Line↗

PubMed1998

The molecular and cellular biology of pancreatic cancer.

Adenocarcinoma of the pancreas carries a grave prognosis for affected patients. Certain oncogenes (K-ras and HER-2/neu) are mutated in a large proportion of these aggressive tumors. Adenocarcinoma of the pancreas has also been associated with loss of tumor suppressor genes (p53, DPC4, p16/MTS), either by deletion or by mutation and loss of function. Growth factors (EGF, TGF-alpha, HGF) and growth factor receptors (EGF-R, c-met, CCK) are expressed at levels not found in the normal pancreas. Finally, factors important for angiogenesis (FGF, integrins, selectins) are likely to play an important role in the growth and metastasis of clinically relevant tumors. This review attempts to summarize and assimilate current research into the molecular and cellular biology of pancreatic cancer.

Adenocarcinoma↗

PubMed1997

Gastrointestinal complications following cardiac surgery. An analysis of 1477 cardiac surgery patients.

OBJECTIVE: To determine preoperative and perioperative risk factors for gastrointestinal (GI) complications following cardiac surgery. DESIGN: A database including records of patients who underwent cardiac surgery was reviewed, with univariate analysis of several variables thought to be relevant to GI complications. Using a risk-adjusted model, preoperative stratification was used to fit a logistic regression model including operative features. SETTING AND PATIENTS: All patients undergoing cardiac surgery from January 1, 1991, to December 31, 1994, at a university-affiliated teaching hospital. MAIN OUTCOME MEASURES: Incidence of GI complications, postoperative mortality, length of hospital stay, and relative risk of GI complications based on multivariate analyses. RESULTS: Gastrointestinal complications occurred in 2.1% of patients and had an associated mortality of 19.4%; this was higher than the mortality in patients without GI complications (4.1%; P < .001). Length of hospital stay was significantly longer in patients with GI complications (43 vs 13.4 days; P < .001). In patients who underwent coronary artery bypass grafting only, cardiopulmonary bypass time was significantly longer in patients with GI complications (166 vs 138 minutes; P = .004). In patients who underwent valve replacement, bypass time was not associated with GI complications. Use of a left internal mammary artery graft was associated with a lower incidence of GI complications. CONCLUSIONS: Patients who have GI complications after cardiac surgery have a higher mortality and a longer hospital stay. The use of a left internal mammary artery seems to have a protective effect against GI complications. Based on these observations, patients may be stratified into low-, medium-, and high-risk groups.

Aged↗

PubMedDate not supplied

26S proteasome inhibition induces apoptosis and limits growth of human pancreatic cancer.

The 26S proteasome degrades proteins that regulate transcription factor activation, cell cycle progression, and apoptosis. In cancer, this may allow for uncontrolled cell division, promoting tumor growth, and spread. We examined whether selective inhibition of the 26S proteasome with PS-341, a dipeptide boronic acid analogue, would block proliferation and induce apoptosis in human pancreatic cancer. Proteasome inhibition significantly blocked mitogen (FCS) induced proliferation of BxPC3 human pancreatic cancer cells in vitro, while arresting cell cycle progression and inducing apoptosis by 24 h. Accumulation of p21(Cip1-Waf-1), a cyclin dependent kinase (CDK) inhibitor normally degraded by the 26S proteasome, occurred by 3 h and correlated with cell cycle arrest. When BxPC3 pancreatic cancer xenografts were established in athymic nu/nu mice, weekly administration of 1 mg/kg PS-341 significantly inhibited tumor growth. Both cellular apoptosis and p21(Cip1-Waf-1) protein levels were increased in PS-341 treated xenografts. Inhibition of tumor xenograft growth was greatest (89%) when PS-341 was combined with the tumoricidal agent CPT-11. Combined CPT-11/PS-341 therapy, but not single agent therapy, yielded highly apoptotic tumors, significantly inhibited tumor cell proliferation, and blocked NF-kappaB activation indicating this systemic therapy was effective at the cancer cell level. 26S proteasome inhibition may represent a new therapeutic approach against this highly resistant and lethal malignancy.

Adenocarcinoma↗

PubMedDate not supplied

Sodium salicylate inhibits proliferation and induces G1 cell cycle arrest in human pancreatic cancer cell lines.

The mutations most common in pancreatic cancer decrease the ability to control G1 to S cell cycle progression and cellular proliferation. In colorectal cancer cells, nonsteroidal anti-inflammatory drugs inhibit proliferation and induce cell cycle arrest. We examined whether sodium salicylate, an aspirin metabolite, could inhibit proliferation in human pancreatic cancer cell lines (BxPC3 and Panc-1). Quiescent cells were treated with medium containing 10% fetal calf serum, with or without salicylate. Cellular proliferation was measured by MTT assay and bromodeoxyuridine incorporation. The fractions of cells in G0/G1, S, and G2/M phases of the cell cycle were quantitated by fluorescence-activated cell sorting. Results were compared between groups by two-tailed t test. Cyclin D1 expression was determined by Western blot analysis and prostaglandin E2 expression by enzyme-linked immunosorbent assay. Serum-starved cells failed to proliferate, with most arrested in the G1 phase. Salicylate significantly inhibited serum-induced progression from G1 to S phase, cellular proliferation, and the expression of cyclin D1. The concentrations at which 50% of serum-induced proliferation was inhibited were 1.2 mmol/L (Panc-1) and 1.7 mmol/L (BxPC3). The antiproliferative effect of sodium salicylate was not explained by inhibition of prostaglandin E2 production. This study provides further evidence in a noncolorectal cancer model for the antineoplastic effects of nonsteroidal anti-inflammatory drugs.

Adenocarcinoma↗