The combination of HSV-tk and endostatin gene therapy eradicates orthotopic human renal cell carcinomas in nude mice

BACKGROUND AND METHODS: Gene therapy may offer a new tool for the treatment of renal cell carcinoma (RCC). We have tested a combination of cytotoxic and antiangiogenic gene therapy for wild-type orthotopic human RCC xenografts in nude mice using intratumoral adenovirus-mediated herpes simplex virus thymidine kinase (HSV-tk) and endostatin (ES) gene therapy. In vivo magnetic resonance imaging, morphometry, immunocytochemistry, and survival were used to evaluate the treatment effect. Adenovirus-mediated marker gene transfers (GFP) were used as controls. Results: In vivo transduction efficiency, measured using GFP gene transfer, was 27+/-7%. The combination gene therapy with HSV-tk and ES adenoviruses resulted in a significant antitumor effect (P<.01) compared to single HSV-tk (n.s.) or ES (n.s.). In the survival study, all tumors with single gene therapy using HSV-tk, ES, and marker gene adenoviruses showed progression in magnetic resonance imaging. In contrast, the majority of the tumors in the combination treatment group remained dormant or were eradicated (57%). Survival of these mice equaled healthy nude mice, and was significantly prolonged (P<.0001) compared to HSV-tk (P<.028) and ES (n.s.) groups. Conclusions: It is concluded that the inhibition of angiogenesis using ES gene transfer together with the cytotoxic HSV-tk gene therapy results in a significantly improved treatment effect in RCC compared to the single gene treatments.     

Haemophilus influenzae type b (Hib) antibody concentrations and vaccination responses in patients with chronic lymphocytic leukaemia: predicting factors for response

We have recently demonstrated a moderate vaccination response rate of 43% against Haemophilus influenzae type b (Hib) conjugate vaccine among adult and elderly patients with chronic lymphocytic leukaemia (CLL). We now investigated demographic and immunological factors predicting the favourable response and protective antibody concentrations for Hib conjugate vaccine in CLL. Lower age was associated with protective pre- and post-vaccination antibody concentrations. High IgG1 and IgA concentrations were also associated with the protective efficacy. High IgM, in turn, was the best predictor of a significant vaccination response. Again, lower age seemed to be involved in this outcome. Judging from these findings, it would seem beneficial to vaccinate all CLL patients with conjugate vaccines at the presentation of the disease. Investigations of a new pneumococcal conjugate vaccine in CLL are warranted.     

Relationships of in vitro sensitivities tested with nine drugs and two types of irradiation in chronic lymphocytic leukemia

Extensive research into mechanisms of cytotoxic drug and irradiation resistance have produced few clinically encouraging results. In this report, we apply correlation analyses to drug and irradiation response results from a cohort of 36 classical B chronic lymphocyte leukemia (CLL) patients. Nine drugs and two types of irradiation were selected according to their usefulness in CLL therapy or on the basis of their otherwise interesting mechanisms of action. Part of the results concerning individual drugs have been previously published, but new correlation analyses are presented in this paper. Altogether 2376 duplicate cultures were performed in order to determine ID(80) values, i.e. doses causing an 80% inhibition in 4-day cultures when leucine incorporation was used as an indicator of cells vitality. Non-parametric Spearman's rank order correlation confirmed a tight relationship between 2-chlorodeoxyadenosine and fludarabine, as expected. Surprisingly, correlation between two P-glycoprotein-dependent drugs, vincristine and doxorubicin, was not demonstrable. A number of entirely unexpected correlations were identified between drugs with very different mechanisms of action: (i) chlorambucil and gamma-irradiation; (ii) 2-chlorodeoxyadenosine and vincristine; (iii) 2-chlorodeoxyadenosine and gamma-irradiation; (iv) fludarabine and cis-platin; (v) doxorubicine and gamma-irradiation; (vi) prednisolone and cyclosporin A; (vii) vincristine and verapamil. Our findings emphasize: (i) the usefulness of fresh tumor cells instead of cell lines in cytotoxicity studies; (ii) the great variation in cytotoxicity in individual patients, i.e. tumor cell heterogeneity, as well as patient heterogeneity; and (iii) an entirely unexpected finding that there were tight relationships in drug and irradiation responses between substances supposed to act with very different mechanisms.     

MR perfusion, diffusion and BOLD imaging of methotrexate-exposed swine brain

PURPOSE: To evaluate the methotrexate (MTX)-exposed swine brain, functional magnetic resonance imaging (MRI), including perfusion, diffusion, and blood-oxygen-level-dependent (BOLD) contrast imaging, was used. MATERIAL AND METHODS: Juvenile pigs received either 2 x 5 g/m(2), or 5 x 2 g/m(2) MTX intravenously within one month. MRI was performed (sedative: propofol) before (14-17 kg, N = 6) and after (21-27 kg, N = 4) the MTX exposure. Also, age-matched controls (22-27 kg, N = 4) were imaged. RESULTS: After the MTX exposure, reduced (from 2%-4% to 0%-1%) or negative (-2% to -3%) BOLD responses were detected; apparent diffusion coefficient (ADC) or relative perfusion values did not change. CONCLUSION: This study suggests that MTX-related changes in the brain may be detected as changes in flow-metabolism coupling as reduced or negative response (for somatosensory activation) in the BOLD contrast MRI. The contrast agent perfusion MRI, without absolute quantification, may not show global damage in brain perfusion related to the MTX exposure in the swine model used. ADC (in one direction) may not indicate MTX-related changes in the brain.     

PATHOLOGY OF LETHAL FETAL GROWTH RETARDATION SYNDROME WITH AMINOACIDURIA, IRON OVERLOAD, AND LACTIC ACIDOSIS (GRACILE)

Autopsy study of 17 newborn infants with lethal autosomal recessive disease presenting as growth retardation with lactic acidosis, Fanconi aminoaciduria, and hepatic hemosiderosis is reported. The patients succumbed between day 1 and 4 months of life; 9 patients died within the first month. All patients showed severe pathologic changes of liver with cholestasis in all livers. Extensive accumulation of stainable iron of the hepatocytes was present in 9/17 autopsy tissues and in two biopsy specimens. Moderate to abundant iron storage in the Kupffer cells was seen in all liver specimens. The amount of hepatocytic iron was high in livers up to 1 month of age and decreased thereafter. The general features and liver findings of this disorder suggest the name Growth Retardation Aminoaciduria Cholestasis Iron Overload, Lactacidosis and Early Death (GRACILE, OMIM 603358). Calcified concrements were seen in the medulla of 13/16 kidney specimens. Pancreas of 13/14 patients showed interstitial fibrosis and exocrine atrophy. Various pathologic findings such as renal tubular dysgenesis, paucity of hepatic bile ducts and iron storage in the macrophages of spleen and pulmonary alveoli were observed in some cases. Previous extensive clinical genetic and laboratory investigations have revealed that the patients had a previously unrecognized genetic disease. It is inherited as an autosomal recessive trait. The gene locus is 2q 33-37. The basic defect of the disease remains unknown.     

The effect of ketoprofen on recovery after tonsillectomy in children: a 3-week follow-up study.

OBJECTIVE: To evaluate recovery after tonsillectomy in children, and to determine the safety and efficacy of ketoprofen in pain treatment after discharge. STUDY DESIGN: A prospective, longitudinal study in 102 children undergoing tonsillectomy. METHODS: All children underwent tonsillectomy under a same general anesthesia. At discharge, all patients were prescribed ketoprofen capsules at a dose of 3-5 mg(-1) kg(-1) per 24 h for postoperative pain control at home, with paracetamol or paracetamol-codeine tablets for rescue analgesia. At home, the patients recorded pain and analgesic consumption each day for the first week after surgery. At 3 weeks, patients recorded the total analgesic requirement, duration of pain, and all adverse events during recovery and return to normal daily activities. RESULTS: The median of pain cessation was 9 days (range 1-20 days) and the median duration of analgesic treatment was 10 days (4-19 days). More than 50% of the patients needed rescue analgesic daily during the first week after tonsillectomy. Ketoprofen combined with paracetamol or paracetamol-codeine provided sufficient analgesia for most children. However, the analgesic action of drugs was too short to achieve pain relief, which allow undisturbed sleep during the first postoperative nights. A return back to normal daily activities took place after 9 days (2-26 days). The influence of age for pain pattern was negligible. Five patients needed electrocautery to stop postoperative bleeding. No other serious adverse-events occurred. CONCLUSIONS: The main problem after tonsillectomy is significant pain that may last 9 days or longer after surgery. Ketoprofen combined with paracetamol-codeine seems to provide a sufficient analgesia, but before ketoprofen may be recommended for children during tonsillectomy a larger study is needed to show whether or not ketoprofen increases the hemorrhage rate.     

Loss of time-organized sympathetic skin responses in acute psychosis

The aim of this study was to differentiate the response pattern characteristics of sympathetic skin responses (SSR) between unmedicated first psychotic subjects and healthy controls. We recorded SSR to novel auditory stimuli in 13 psychotic subjects and in 19 controls. There was no constant delay between a sudden change in the acoustic surroundings and SSR in psychotic subjects, whereas regularly this delay was 1.3 to 1.8 seconds in controls. The validity of the test was evaluated by blind rating responses to two categories. The lack of a regular time delay between a novel auditory stimulus and SSR suggests a timing disturbance in the neural networks regulating the autonomic nervous system responses in acute psychosis. Our finding and method may have future implications in evaluation of subtle autonomic nervous system alterations related, e.g., to drug effects.     

Cyst fluid tumor-associated trypsin inhibitor may be helpful in the differentiation of cystic pancreatic lesions

In clinical practice it is important to differentiate pseudocysts from cystic pancreatic tumors, especially potentially malignant mucinous cystic tumors. We investigated three new markers—tumor-associated trypsin inhibitor (TATI) and the free α and β subunits of human choriogonadotropin (hCGα and hCGβ, respectively)—in the cyst.uid of patients with cystic pancreatic lesions and compared the concentrations of these markers to those of carcinoembryonic antigen (CEA), CA 19-9, CA 242, CA 125, CA 15-3, alpha-fetoprotein, and tissue polypeptide antigen in order to distinguish benign cysts from malignant cysts. Between 1995 and 2001, a total of 34 patients operated on for cystic pancreatic lesions at Tampere University Hospital were included. Cyst fluid was aspirated at operation and stored at_70 C. Thehistologic diagnosis was pseudocyst in 23 patients, serous cystadenoma (SCA) in four patients, benign mucinous cystadenoma (MCA) in four patients, cystic papillary neoplasm (CPN) in one patient, glucagonoma in one patient, and malignant endocrine islet cell carcinoma (EC) in one patient. Significantly higher concentrations of TATI were found in patients with MCA and EC (2239 ± 149 μg/L [mean ± SEM]) than in patients with pseudocyst (55 ± 29 μg/L; P = 0.001) and in patients with SCA (36 ± 23 μg/L; P = 0.01). The patient with CPN and the patient with glucagonoma had relatively low levels of TATI (30.7 and 46.5 μg/L). Mean CEA was higher in patients with MCA compared to those with pseudocysts (19,993 ± 9418 vs. 53 ± 20 μg/L, P = 0.002) and SCA (0.4 ± 0.1 μg/L; P = 0.02), but in the patient with malignant EC, the patient with CPN, and the patient with glucagonoma, CEA was normal. HCGα, hCGβ, CA 19-9, CA 242, CA 125, CA 15-3, alpha fetoprotein, and tissue polypeptide antigen could not distinguish between MCA vs. pseudocyst or SCA, because both normal and elevated values were seen in all groups. To our knowledge, this is the first time that TATI has been quantitated in the cyst fluid of patients with cystic pancreatic lesions. It appears to be a potential marker in the differential diagnosis of benign from malignant cystic pancreatic lesions. Supported by the Medical Research Fund of Tampere University Hospital.     

Tolerability and Kinetics of a Solvent-Detergent-Treated Intravenous Immunoglobulin Preparation in Hypogammaglobulinaemia Patients

The tolerability and kinetics of a solvent-detergent-treated 6% intravenous immunoglobulin (IVIG) preparation were studied in 15 hypogammaglobulinaemia patients during 3–4 regular substitution infusions of 9–18 g, the mean dose being 359 mg/kg. The infusions were well tolerated, and the trough serum IgG levels achieved were comparable to two commercial IVIG preparations. The stepwise increase of the infusion rate up to 5 mg/kg/min and the use of this IVIG as a 12% solution were possible without serious adverse events in all the 6 studied hypogammaglobulinaemia patients. This greatly reduced the time needed for the infusions.     

Failure of hand disinfection with frequent hand washing: a need for prolonged field studies

In a prolonged field trial a 4% chlorhexidine digluconate detergent scrub (Hibiscrub^R), that had earlier proved to be an effective hand disinfectant, was studied in hospital wards. Finger tips were found to harbour more bacteria than the hand dorsum and the samples collected from them yielded more information on the bacteriological and dermatological effects of hand disinfectants in practice.