Wnt-induced, TRP53-mediated Cell Cycle Arrest of Precursors Underlies Interstitial Cell of Cajal Depletion During Aging.

BACKGROUND & AIMS: Gastric dysfunction in the elderly may cause reduced food intake, frailty, and increased mortality. The pacemaker and neuromodulator cells interstitial cells of Cajal (ICC) decline with age in humans, and their loss contributes to gastric dysfunction in progeric klotho mice hypomorphic for the anti-aging Klotho protein. The mechanisms of ICC depletion remain unclear. Klotho attenuates Wnt (wingless-type MMTV integration site) signaling. Here, we examined whether unopposed Wnt signaling could underlie aging-associated ICC loss by up-regulating transformation related protein TRP53 in ICC stem cells (ICC-SC). METHODS: Mice aged 1-107 weeks, klotho mice, APCΔ468 mice with overactive Wnt signaling, mouse ICC-SC, and human gastric smooth muscles were studied by RNA sequencing, reverse transcription-polymerase chain reaction, immunoblots, immunofluorescence, histochemistry, flow cytometry, and methyltetrazolium, ethynyl/bromodeoxyuridine incorporation, and ex-vivo gastric compliance assays. Cells were manipulated pharmacologically and by gene overexpression and RNA interference. RESULTS: The klotho and aged mice showed similar ICC loss and impaired gastric compliance. ICC-SC decline preceded ICC depletion. Canonical Wnt signaling and TRP53 increased in gastric muscles of klotho and aged mice and middle-aged humans. Overstimulated canonical Wnt signaling increased DNA damage response and TRP53 and reduced ICC-SC self-renewal and gastric ICC. TRP53 induction persistently inhibited G1/S and G2/M cell cycle phase transitions without activating apoptosis, autophagy, cellular quiescence, or canonical markers/mediators of senescence. G1/S block reflected increased cyclin-dependent kinase inhibitor 1B and reduced cyclin D1 from reduced extracellular signal-regulated kinase activity. CONCLUSIONS: Increased Wnt signaling causes age-related ICC loss by up-regulating TRP53, which induces persistent ICC-SC cell cycle arrest without up-regulating canonical senescence markers.

Accuracy of maxillary repositioning surgery using CAD/CAM customized surgical guides and fixation plates.

The advent of three-dimensional imaging and computer-aided surgical simulation (CASS) have brought about a paradigm shift in surgical planning. The aim of this study was to assess the accuracy of maxillary repositioning surgery using computer-aided design and manufacturing (CAD/CAM) customized titanium surgical guides and fixation plates. Thirty consecutive adult patients, 13 male and 17 female, with a mean age of 29.2 years and 25.5 years, respectively, requiring Le Fort I maxillary osteotomy, with or without simultaneous mandibular surgery, were evaluated retrospectively. All orthognathic surgeries were performed by one experienced surgeon. The pre-surgical and post-surgical volumetric imaging were superimposed to assess the linear and angular differences between the planned and actual positions of the maxilla following surgery. With the use of the CAD/CAM titanium surgical guides and fixation plates, all surgical movements were within 2mm and 4° of the planned movements, which is considered clinically insignificant. The overall root mean square error between the planned and actual surgical movements was 0.38mm in the transverse dimension, 0.64mm in the anteroposterior dimension, and 0.55mm in the vertical dimension. In regard to the centroid of the maxilla, the absolute angular difference of the maxillary centroid was 1.06° in pitch, 0.47° in roll, and 0.49° in yaw. Maxillary repositioning surgery can be performed with high accuracy using CAD/CAM titanium surgical guides and fixation plates.

Rapid, deep and precise profiling of the plasma proteome with multi-nanoparticle protein corona.

Large-scale, unbiased proteomics studies are constrained by the complexity of the plasma proteome. Here we report a highly parallel protein quantitation platform integrating nanoparticle (NP) protein coronas with liquid chromatography-mass spectrometry for efficient proteomic profiling. A protein corona is a protein layer adsorbed onto NPs upon contact with biofluids. Varying the physicochemical properties of engineered NPs translates to distinct protein corona patterns enabling differential and reproducible interrogation of biological samples, including deep sampling of the plasma proteome. Spike experiments confirm a linear signal response. The median coefficient of variation was 22%. We screened 43 NPs and selected a panel of 5, which detect more than 2,000 proteins from 141 plasma samples using a 96-well automated workflow in a pilot non-small cell lung cancer classification study. Our streamlined workflow combines depth of coverage and throughput with precise quantification based on unique interactions between proteins and NPs engineered for deep and scalable quantitative proteomic studies.

Sapienza Global Bedside Evaluation of Swallowing after Stroke: the GLOBE-3S study.

BACKGROUND AND PURPOSE: Dysphagia occurs in up to 50% of all patients with acute stroke. There is debate regarding which is the most effective screening tool in identifying aspiration in patients with acute stroke. We assessed the accuracy of the Sapienza Global Bedside Evaluation of Swallowing after Stroke (GLOBE-3S), which combines the Toronto Bedside Swallowing Screening Test (TOR-BSST©) with oxygen desaturation and laryngeal elevation measurement during swallowing. METHODS: We prospectively enrolled consecutive patients with stroke within 72 h of symptom onset. All patients with stroke firstly underwent a standard neurological examination, then the GLOBE-3S evaluation and finally the fiberoptic endoscopic evaluation of swallowing (FEES). Two different assessors, a neurologist and a speech pathologist, blind to both the clinical data and each other's evaluation, administered the GLOBE-3S and FEES examination. We assessed the accuracy of the GLOBE-3S in detecting post-stroke swallow impairment with aspiration using the FEES as the standard. RESULTS: We enrolled 50 patients with acute stroke, 28 of whom (56%) had swallowing impairment with aspiration at FEES evaluation. A total of 33 patients (66%) failed the GLOBE-3S evaluation. The GLOBE-3S reached a sensitivity of 100% and a specificity of 77.3% (negative predictive value, 100%; positive likelihood ratio, 4.34). The median time required for the GLOBE-3S to be performed was 297 s. CONCLUSIONS: GLOBE-3S is quick to perform at the bedside and can accurately identify aspiration in patients with acute stroke. By including the measurement of laryngeal elevation and monitoring of oxygen desaturation, it could represent a highly sensitive instrument to avoid the misdiagnosis of silent aspirators.

Complementary medicine use by men with prostate cancer: a systematic review of prevalence studies.

Men with prostate cancer are reported as commonly using complementary and alternative medicine (CAM) but surveys have not recently been subjected to a rigorous systematic review incorporating quality assessment. Six electronic databases were searched using pre-defined terms. Detailed information was extracted systematically from each relevant article. Study reporting quality was assessed using a quality assessment tool, which demonstrated acceptable inter-rater reliability and produces a percentage score. In all, 42 studies are reviewed. All were published in English between 1999 and 2009; 60% were conducted in the United States. The reporting quality was mixed (median score = 66%, range 23-94%). Significant heterogeneity precluded formal meta-analysis. In all, 39 studies covering 11,736 men reported overall prevalence of CAM use; this ranged from 8 to 90% (median=30%). In all, 10 studies reported prevalence of CAM use specifically for cancer care; this ranged from 8 to 50% (median = 30%). Some evidence suggested CAM use is more common in men with higher education/incomes and more severe disease. The prevalence of CAM use among men with prostate cancer varies greatly across studies. Future studies should use standardised and validated data collection techniques to reduce bias and enhance comparability.

An 18-year review of sports groin injuries in the elite hockey player: clinical presentation, new diagnostic imaging, treatment, and results.

OBJECTIVE: This is a retrospective study of 98 hockey players who underwent 107 surgical explorations for refractory lower abdominal and groin pain that prevented them from playing hockey at an elite level. DESIGN: Retrospective chart review combined with a complete follow-up examination and questionnaire. SETTING: The players were treated in an ambulatory care university tertiary care centre. PATIENTS: A total of 98 elite hockey players underwent 107 surgical groin explorations for intractable groin pain preventing their play. Follow-up was 100%. INTERVENTION: Each player had repair of a tear of the external oblique muscle and fascia reinforced by a Goretex mesh. The ilioinguinal nerve was resected in each patient. OUTCOME MEASURES: There was absence of groin pain on the return to play hockey at an elite level. RESULTS: In all, 97 of 98 players returned to play after the surgical procedures. No morbidity was attributed to division of the ilioinguinal nerve. CONCLUSIONS: Surgical exploration of the involved groin with repair of the torn external oblique muscle and division of the ilioinguinal nerve has resulted in resolution of refractory groin pain and return to play in the elite hockey player. The surgical procedure is associated with a low morbidity. Recent observations on dynamic ultrasound show promise in accurately diagnosing this injury.

An expanding role for cardiopulmonary bypass in trauma.

OBJECTIVES: To analyze experience at the McGill University Health Centre with cardiopulmonary bypass (CPB) in trauma, complemented by a review of the literature to define its role globally and outline indications for its expanded use in trauma management. DATA SOURCES: All available published English-language articles from peer reviewed journals, located using the MEDLINE database. Chapters from relevant, current textbooks were also utilized. STUDY SELECTION: Nine relevant case reports, original articles or reviews pertaining to the use of CPB in trauma. DATA EXTRACTION: Original data as well as authors' opinions pertinent to the application of CPB to trauma were extracted, incorporated and appropriately referenced in our review. DATA SYNTHESIS: Overall mortality in the selected series of CPB used in the trauma setting was 44.4%. Four of 5 survivors had CPB instituted early (first procedure in operative management) whereas 3 of 4 deaths involved late institution of CPB. CONCLUSIONS: Although CPB has traditionally been used in the setting of cardiac trauma alone, a better understanding of its potential benefit in noncardiac injuries will likely make for improved outcomes in the increasingly diverse number of severely injured patients seen in trauma centres today. Further studies by other trauma centres will allow for standardized indications for the use of CPB in trauma.

Tailored therapy for aggressive non-Hodgkin's lymphoma: results of a phase II study with a long-term follow-up.

Aim of the study was to improve cure rate and survival of aggressive non-Hodgkin's lymphoma (NHL) with a tailored program of therapy based on histologic type, prognostic characteristics of patients and response to therapy, and with the use of differentiating or cytostatic agents such as Ara-C at low doses and alphaIFN. Fifty-four consecutive patients with aggressive NHL were treated in the induction phase with 4 sequential courses of a third generation regimen (modified CODBLAM IV), followed in responsive patients by 1 cycle of doxorubicin and cyclophosphamide and 1 cycle of high dose methotrexate with folinic acid rescue (AC-MTX). Patients who achieved partial response (PR) were treated with the combination of CCNU + vinblastine if affected by high grade NHL, or with low dose Ara-C plus alphaIFN if affected by intermediate grade NHL. Patients who obtained complete response (CR) with basal adverse prognostic factors were treated with alphaIFN as maintenance therapy for two years. Radiotherapy and surgery were effected in selected cases. Thirty-four patients (62.9%) achieved CR and 12 patients (22.2%) showed PR after induction therapy. Among the 12 patients who achieved PR, 6 prolonged CRs were obtained in 7 patients treated with Ara-C at low doses plus alphaIFN and 4 CRs were obtained in 5 patients treated with CCNU + vinblastine. After completion of treatment, 44 patients (81.5%) obtained CR, 2 patients (3.7%) showed PR and 8 patients (14.8%) presented progression of disease (PD). Fifteen patients received alphaIFN as maintenance therapy. The overall survival and failure-free survival rates are 53.7% and 50% respectively, with a median follow-up of 82 months: 27 patients remain alive, disease-free without relapses, and can be considered cured. This tailored program of therapy resulted effective and moderately toxic and may improve the outcome in aggressive NHL.

Thymostimulin treatment of hepatocellular carcinoma on liver cirrhosis.

Hepatocellular carcinoma (HCC) on liver cirrhosis (LC) is a neoplasm with a poor prognosis. Only 10-36% of patients have resectable tumors at presentation, systemic chemotherapy is seldom useful and loco-regional treatments are feasible only in selected cases. On the basis of the possible role of depression of immune function in the pathogenesis of HCC on LC, we employed thymostimulin in the treatment of patients with unresectable HCC on LC, with the aim of influencing the course of the disease. Forty-six consecutive patients with HCC were enrolled from January 1987 to December 1993. All patients, 36 males and 10 females, with a median age of 67 years (range 58-82) had coexisting LC. All patients were treated with thymostimulin (TP-1 Serono) at doses of 70 mg/day i.m. five times a week until disease progression. Eleven patients achieved an objective response, with an overall response rate of 24%. Six patients obtained a CR (13.0%) with a median duration of 19+ months (range 5+/-23). Five patients (11%) achieved PR with a median duration of 10 months (range 7+/-22). Six patients (13.0%) presented MR (median duration: 12.5 months) and 12 (26.0%) had SD (median duration: 8 months). In 17 patients (36.9%) the disease progressed. Thymostimulin treatment was well tolerated and no major side effect was observed. Thymostimulin treatment can be considered a new promising therapy for advanced HCC.

Could thymostimulin prevent hepatocellular carcinoma occurrence in patients with liver cirrhosis?

On the basis of a possible pathogenetic role of immunodepression in the development of hepatocellular carcinoma in patients with chronic hepatitis B/C viral infection and liver cirrhosis, we treated 34 liver cirrhosis patients (M/F 24/10; age: 14 pts less than or equal to 60 years, 20>60; Performance status: 22 0-1, 12 2-3; Childs' grade: 21 A, 13 B; ascites yes/not: 4/30) employing thymostimulin (TST), at the dosage of 50 mg/m(2) i.m. 3 times a week, until death or severe toxicity occurrence. Etiology of cirrhosis was viral in all cases. The patients were followed up every 3 months by means of clinical examination and biochemical analyses; every 6 months by checking viral serum markers, alpha-fetoprotein and by means of ultrasounds. To date, 34 patients have been treated, with a median follow-up of 6 (1-8) years. No occurrence of hepatocellular carcinoma has been observed, with a statistically significant difference between observed and expected HCC (p<0.05). Thymostimulin treatment was well tolerated. Our data seem to suggest that the immunomodulating treatment could significantly reduce the risk of HCC occurrence in patients with liver cirrhosis.

Maintenance therapy with recombinant interferon alpha-2B (alpha IFN) in prognostically unfavourable aggressive non-Hodgkin's lymphomas (NHL).

Fifteen patients from a series of 37 consecutive patients with aggressive NHL who achieved a complete remission (CR) with first line chemotherapy were defined at high or high-intermediate risk of relapse according to International Prognostic Risk Index and were treated with alpha IFN as maintenance therapy for two years. After a median follow-up of 62 months only 3/15 patients (20%) relapsed and 2/15 (13.3%) died in the alpha IFN treated group, favourably comparing with the expected relapse and death rate in that setting; on the other hand 40.9% of 'low risk' patients from the same series, who did not receive alpha IFN, had relapsed. alpha IFN maintenance therapy appears to result in prolonged response duration and survival in patients with aggressive and prognostically unfavourable NHL.

Relationship between icam-1 serum levels and thymostimulin therapy in patients with liver-cirrhosis associated with hepatocellular-carcinoma.

Intercellular adhesion molecule-1 (ICAM-1), is a cell surface molecule involved in many immunological processes. Recently, ICAM-1 has also been detected as a soluble form in sera of patients affected by different pathologies and often associated with severity of the disease. However, all these studies were carried out in patients before drug treatment. Since in vitro experiments and recent evidence in humans has indicated the possibility that some cytokines may be responsible of ICAM-1 release, we have considered the possibility of quantitating ICAM-1 in sera of patients undergoing therapy with drugs influencing cytokine regulation. Thymostimulin, which seems to belong to this category, has been used in a pilot study by some of us in the treatment of patients with liver cirrhosis associated with hepatocellular carcinoma. By using an enzymatic sandwich assay, we found significantly high circulating ICAM-1 mean values in patients affected by liver cirrhosis alone or associated with hepatocellular carcinoma or non-Hodgkin lymphoma compared to control subjects (p<0.001), Moreover, in patients with hepatocellular carcinoma or liver cirrhosis who were treated with thymostimulin, serial detection of c-ICAM-1 was carried out and patients responsive to the drug showed significantly increased values of c-ICAM-1 (p=0.006), Our study suggests that an evaluation of c-ICAM-1 levels needs to be considered in the context of the therapeutic approach, with particular attention to those drugs which may affect the regulation of the cytokine pathway.

Mitoxantrone, Etoposide and bleomycin (meb) chemotherapy in non-hodgkins-lymphoma patients non-elegible for standard cyclophosphamide, Doxorubicin, vincristine and prednisone (chop) combination.

The antitumor activity and toxicity profile of a new therapeutic combination was investigated for patients with non-Hodgkin's lymphoma (NHL). The regimen consisted of mitoxantrone (10 mg/m(2)/day by intravenous (i.v.) bolus injection on day 1), etoposide (100 mg by 24 hours continuous i.v. infusion on days 1, 2, 3) and bleomycin (4 mg by i.v. bolus injection on day 1 followed by 24 hours continuous i.v. infusion at 4 mg/m2/day dose on days 1, 2, 3) (MEB). MEB chemotherapy was administered to 22 patients affected by intermediate/high grade or clinically symptomatic low grade NHL who were considered non-elegible for standard cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) chemotherapy. Major responses were achieved in 11/22 (50%) patients with 5 (23%) complete responses. Grade 3-4 neutropenia occurred in 59% of patients. The results of this study demonstrate that MEB chemotherapy possesses good antitumor activity and a manageable toxicity in a prognostically unfavourable subset of lymphoma patients.

Cytosine arabinoside (Ara-C) plus alpha-interferon (alpha IFN) determine prolonged complete remissions in patients with aggressive non-Hodgkin's lymphoma partially responsive to first-line doxorubicin-containing regimens.

Ten patients with aggressive NHL who failed to achieve a complete remission with first-line chemotherapy were treated with Ara-C and alpha IFN. Ara-C was administered subcutaneously at 100 mg on day 1, 150 mg on day 2 and 200 mg on days 3, 4 and 5 of a 28 d cycle; alpha IFN was given at 3 million International Units three times a week and continued for 2 years in CR patients. Six CR were attained with a median duration of 36+ months. Toxicity was mild. A new approach to second-line therapy for aggressive NHL is proposed.

The enterocyte-like differentiation of the Caco-2 tumor cell line strongly correlates with responsiveness to cAMP and activation of kinase A pathway.

We have investigated the expression of protein kinase C (PKC) and protein kinase A (PKA) during the phases of growth and differentiation of the human colon carcinoma Caco-2 cells. We studied whether differentiation correlated with the responsiveness to cAMP and with an increased transport of the catalytic subunit of PKA into the nucleus. Also, we evaluated whether this phenomenon was affected by PKC activity. High levels of activated PKC were found in the plasma membranes of replicating cells. When the cells began to differentiate, plasma membrane-activated PKC decreased, while the cytosolic fraction increased. On the contrary, PKA holoenzyme increased during differentiation, along with the transport of its catalytic subunit into the nucleus. Both types I and II kinase A holoenzymes increased during differentiation, with maximal type II activity found when cells were fully differentiated. In replicating preconfluent cells, the inhibition of PKC by high dose phorbol 12-myristate 13-acetate or sphingosine increased the amount of both PKA catalytic subunit in the nucleus and sucrase activity. During differentiation, 8-Bromo-cAMP increased PKA catalytic subunit in the nucleus and apoliprotein A1 mRNA levels. These effects were inhibited by low-dose phorbol 12-myristate 13-acetate, which activates PKC in the plasma membranes. Our data suggest that PKC is activated in proliferating Caco-2 cells. The inhibition of PKC induces the transport of PKA catalytic subunit into the nucleus and the expression of the differentiation markers. Differentiated Caco-2 cells show a lower activation of PKC and an increased transport of the catalytic subunit of PKA into the nucleus.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiovascular monitoring of drug-resistant lymphoma patients treated with epoch chemotherapy plus high-dose verapamil in continuous-infusion.

High dose Verapamil (VP) infusion has been incorporated into cytotoxic chemotherapy in order to circumvent tumor cell drug-resistance. We have evaluated the cardiovascular side-effects produced by high dose VP associated to EPOCH chemotherapy in 12 patients with chemorefractory lymphoma. Continuous monitoring of right ventricular and pulmonary pressure and cardiac index was performed in three patients by a Swan-Ganz catheter. A slight reduction in cardiac index was observed 6 h after the beginning of VP infusion and was followed by spontaneous recovery within 12 h. First degree atrioventricular (AV) block was detected in 6/12 patients. Premature Ventricular Beats (PVB) occurred in one patient, and promptly disappeared after xylocaine administration. All patients experienced mild and transient hypotension, while severe hypotension was observed only in 1 patient, who promptly recovered when VP administration was discontinued. Hypokalemia was detected in 6 patients possibly as a consequence of transient activation of the renin-angiotensin system.

90k is a serum marker of poor-prognosis in non-hodgkins-lymphoma patients.

Monoclonal antibody SP-2, which binds to a 90,000 daltons tumor-associated antigen termed 90K, was generated by mouse immunization with proteins released by human breast cancer cells into the culture medium (Iacobelli et al: Cancer Res 46: 3005-3010, 1986). Elevated 90K levels have been previously reported in the serum of patients with various malignancies. We investigated whether the circulating levels of 90K antigen might be related to prognosis of patients with Non-Hodgkin's Lymphomas (NHL). Serum samples were obtained from 50 apparently healthy blood donors and 81 patients with NHL. Circulating serum 90K concentrations (U/ml) were determined by a solid-phase immunoradiometric assay (IRMA) by a two-step procedure. Serum 90K levels were significantly higher in patients with NHL than in healthy controls (p=0.004). The Kaplan-Meier analysis of overall survival showed that patients with 90K-negative (serum 90K levels less than or equal to 16 U/ml) survived longer than patients with 90K-positive sera (less than or equal to 16 U/ml) (p=0.004). Multivariate regression analysis revealed that serum levels of LDH and 90K were the two independent prognostic variables for predicting overall survival. We propose that an elevated 90K antigen level in sera is a predictor of poor prognosis in NHL.

Low-dose dopamine induces early recovery of recombinant interleukin-2--impaired renal function.

Recombinant interleukin-2 (rIL-2) can produce impairment of renal function with hypotension, fluid retention, elevated blood urea nitrogen, oliguria and low fractional sodium excretion; these side-effects are a common cause of reduction or interruption of rIL-2 infusion. The aim of this study was to investigate the control and treatment of renal toxicity induced by rIL-2 therapy. Here we show that dopamine, at a low dose of 2 micrograms/kg/min, completely prevented renal toxicity induced by rIL-2. While continuing rIL-2 therapy, 24-h continuous infusion of low-dose dopamine produced a rapid normalisation of urine output and a significant decrease in serum creatinine levels and body weight (P < 0.01), with an early and complete recovery of the rIL-2--impaired renal function: mean recovery time of renal function in patients treated with dopamine was significantly lower (P < 0.05) than in nontreated patients (4.8 days vs. 10 days, respectively).