Maxizymes and small hairpin-type RNAs that are driven by a tRNA promoter specifically cleave a chimeric gene associated with leukemia in vitro and in vivo

Many leukemias result from chromosomal translocations that lead to the generation of chimeric oncoproteins. Chimeric gene for the promyelocytic leukemia-retinoic acid receptor alpha (PML-RAR alpha) is the hallmark of acute promyelocytic leukemia. Specific gene silencing of an oncogene is desirable for the treatment of these diseases. We have previously constructed an allosterically controllable ribozyme (designated maxizyme) targeted for bcr-abl chimeric mRNA, whose cleavage activity is induced only in the presence of a specific RNA sequence of interest. It has been demonstrated recently that RNA interference induced by short hairpin-type RNAs provides a powerful method for sequence-specific gene silencing. Here we report that DNA vector-based maxizymes and short hairpin-type RNAs driven by the promoter of a human gene for tRNA(Val) specifically inhibit the expression of the chimeric gene for PML-RAR alpha both in vitro and in vivo. Our findings confirm the potential utility of maxizymes and shRNAs as therapeutic agents.     

Cadmium-induced testicular damage in a rat model of subchronic intoxication

Background: Cadmium (Cd)-induced testicular damage in relation to spermatogenesis has not been well studied. We studied the mechanism of Cd-induced testicular damage in a rat model of subchronic intoxication.
Methods:  Male Sprague–Dawley rats were subcutaneously injected with 0.6 mg Cd/kg per day for 6 weeks. The concentration of Cd in urine, serum and testes was measured by using atomic absorption spectrophotometry. Testicular damage was evaluated by counting the spermatogonia (SG) and spermatocytes (SC) on one cut-surface of five seminiferous tubules in stages VII or VIII of spermatogenesis every week. The location of intratesticular cadmium was determined by using oxine-fluorescent cytochemistry.
Results: There were no differences in the testes/bodyweight ratio between the study and control groups. The concentration of Cd in the testes increased more than 100-fold that in serum after week 2, suggesting active testicular Cd accumulation (1–3 mg/g tissue). Cadmium accumulation was detected in SG and SC. The number of SG and SC diminished significantly in the study group (week 2: SG 74%, SC 90%; week 4: SG 47%, SC 75%; week 6: SG 30%, SC 54% of the control, respectively).
Conclusions: Cadmium accumulated in SG and SC, consequently reduced the number of these cells, and disturbed the spermatogenesis in this rat model of subchronic Cd intoxication. Therefore, the number of SG decreased in this rat model of subchronic Cd intoxication. (Reprod Med Biol 2002; 1 : 59–63).     

Intraocular biodistribution of mono-L-aspartyl chlorin e6 in a primate choroidal neovascularization model.

BACKGROUND AND OBJECTIVE: To describe the biodistribution of a hydrophilic sensitizer, mono-L-aspartyl chlorin e6 (NPe6), in a primate model of choroidal neovascularization (CNV). MATERIALS AND METHODS: NPe6 (25 mg/kg) and indocyanine green (ICG, 15 mg/kg) were intravenously administered simultaneously in monkeys with laser-induced CNVs. Eyes were enucleated and examined by fluorescence microscopy immediately, 20 minutes, and 1, 4, and 24 hours after dye injection. RESULTS: Fluorescence peaked in retinal vessels immediately after dye administration; rapid washout began 1 hour postinjection. In contrast, experimental CNV demonstrated little fluorescence immediately after injection, with increased intensity at later time points. Peak dye fluorescence in CNV occurred 1 hour following dye injection. NPe6 fluorescence diminished rapidly and washed out completely by 24 hours. The dye accumulation and retention pattern of ICG dye resembled that of NPe6. CONCLUSIONS: This study demonstrated the selective accumulation of NPe6 photosensitizer and ICG dye within experimental CNV and rapid clearance from the circulation and retinochoroidal tissue.     

A congenital anterior diaphragmatic hernia with massive pericardial effusion requiring neither emergency pericardiocentesis nor operation. A case report and review of the literature

All previously reported cases of anterior, diaphragmatic hernia with massive pericardial effusion were treated by pericardiocentesis and radical surgery during the early neonatal period. However, we initially followed the course of our patient in the neonatal period. Subsequently, elective surgery was performed at 70 days of age. Including our case, cardiac tamponade has not been observed in any previously reported cases of congenital anterior diaphragmatic hernia with massive pericardial effusion. CONCLUSION: Emergency pericardiocentesis and surgery are not always required immediately after birth, even when the presence of this condition is suspected by prenatal diagnosis. Our observation may be beneficial to preterm low birth weight infants with this condition.     

Cryopreservation of human sperm in patients with malignancy: First 2 years’ experience

Background  Patients with malignancy (n = 130) participated in the sperm cryopreservation program. Methods  After washing and concentrating, sperm was cryopreserved using KS-VIm cryoprotectant medium. Participant background factors such as age, marital status, underlying disease, presence or absence of previous treatment and semen findings (concentration, motility and morphology) were analyzed to determine parameters associated with the program. Results  Patients in their 20s were most common (64 cases) and 94 cases were unmarried at the first visit. The main underlying diseases were testicular tumor (53 cases), leukemia (43 cases) and malignant lymphoma (13 cases). The program was completed for 118 cases. For leukemia, all semen parameters were closer to normal in patients without previous treatment (untreated group, UG) compared with the treated group (TG). When semen findings in the UG were classified according to underlying disease, sperm concentration was lower in patients with testicular tumor compared with those who had leukemia or malignant lymphoma. Four couples underwent reproductive therapies with the cryopreserved sperm through assisted reproductive technology, and three babies were born to two couples. Conclusion  Sperm cryopreservation liberates patients with malignancy from iatrogenic infertility as a consequence of intensive therapy, allowing them to retain reproductive ability.     

Diagnosis, assessment, and treatment of alcohol withdrawal syndrome]

Alcohol withdrawal syndrome is the one of the most critical status among alcohol related disorders. Early detection, diagnosis, and appropriate treatment of alcohol withdrawal syndrome are keys to a successful outcome. This article describes the concrete method of diagnosis, assessment, and treatment of alcohol withdrawal syndrome.     

Changes in the clinical picture of Fukuyama type. Congenital muscular dystrophy in its advanced stage: effectiveness of mechanical ventilation systems

We studied the clinical courses of Fukuyama type congenital muscular dystrophy (FCMD) based on a long-term follow-up of 7 patients. Their peak motor function varied from controlling the head incompletely to standing with support. Six patients had epilepsy, which in some cases was diagnosed with difficulty because of changing seizure types and multiple joint contracture. Six patients underwent endotracheal intubation, and in 5 of them long-term mechanical ventilation began at the age of 15 to 20 years. Three of them underwent trachestomy intermittent positive pressure ventilation (TIPPV), whereas recently 2 patients were treated successfully with nasal intermittent positive pressure ventilation (NIPPV). Mechanical in-exsufflation (MI-E) was effective in clearing airway secretion of 4 patients. With disease progression, the motor, mental, nutritional, cardiorespiratory and swallowing functions of FCMD patients should be assessed to ensure adequate management by NIPPV and MI-E.     

Inhalation of platelet-activating factor increases respiratory resistance in rats: Determination by means of an astograph under nonanesthetized conditions

Intratracheal administration of platelet-activating factor (PAF) to dogs, baboons, and humans has been shown to induce hyperreactivity of the airways and contraction of the smooth muscle. However, it has not yet been reported whether intratracheal administration of PAF to rats induces hyperreactivity. In the present study, the authors estimated the respiratory resistance of rats during intratracheal administration of PAF in order to evaluate the reactivity of the airways to PAF. In both the nonsensitized group and the sensitized group of rats, intratracheal administration of PAF induced an increase in respiratory resistance. The results obtained clarify that responsiveness to PAF exists in the airways of rats.     

Long-term results after femoral vein valve repair for chronic venous insufficiency

OBJECTIVE: Our long-term results with anterior venous sinus plication (AVSP) for femoral vein reconstruction will be presented. PATIENTS AND METHODS: Between 1986 and 2001 we treated 2 100 patients in our hospital for chronic venous insufficiency. In 3.3 % of the patients (n = 70) an AVSP of the target valve, which is the highest valve of the femoral vein distally of the profundal vein branch was carried out. 50 patients could be followed for 2-15 (average 4.6) years postoperatively by phlebographic control. RESULTS: Four recovery patterns after valve repair were seen on venography. The most typical type was the plicated site stop seen in 22 of 50 patients (44 %). Here the venous reflux was stopped at the plicated site directly. The clinical results were good or excellent for all patients. No patient underwent a second procedure for recurrence of the symptoms. A profundal femoral vein reflux did not negatively influence patient outcome. CONCLUSION: AVSP is an excellent method of valve repair in strictly selected patients with chronic venous insufficiency (= no postthrombotic syndrome, no thrombotic occlusion of the femoral veins). Long-term results up to 15 years were highly satisfactory.