Use of subunit-specific antisense oligodeoxynucleotides to define developmental changes in the properties of N-methyl-D-aspartate receptors

Antisense oligodeoxynucleotides were used to determine whether alterations in the expression of N-methyl-D-aspartate (NMDA) receptor subunit mRNA are responsible for developmental changes in the sensitivity of receptors to agonists and antagonists. Xenopus laevis oocytes were injected with mRNA prepared from neonatal and adult rat cerebral cortex, and the effects of agonists and antagonists were determined under voltage-clamp conditions. Glycine-site antagonists like 7-chlorokynurenate and glutamate-site antagonists like CGP-39653 were more potent at NMDA receptors expressed from mRNA from adult rat cerebral cortex than those expressed from mRNA from 1-day-old rat. NMDA receptors from 1-day-old rat cerebral cortex were more sensitive to activation by glycine than were receptors from adult rat cerebral cortex. 7-Chlorokynurenate and CGP-39653 were more potent inhibitors of responses seen with heteromeric NR1/NR2A receptors than with NR1/ NR2B receptors. Conversely, heteromeric NR1/NR2B receptors were more sensitive to activation by glycine than were NR1/NR2A receptors. We previously described a delay in the expression of the NR2A subunit in developing rat brain. Anti-sense oligodeoxynucleotides were used to determine whether the delayed expression of the NR2A subunit underlies changes in pharmacological properties observed during development. The properties of receptors seen when adult brain mRNA was coinjected with antisense oligodeoxynucleotides against the NR2A subunit were similar to those found in receptors from 1-day-old rat brain. These data suggest that changes in the sensitivity of NMDA receptors to antagonists and to glycine seen during development are a result of alterations in the expression of different species of NR2 subunit mRNA.     

Cytomorphologic panorama of cysticercosis on fine needle aspiration. A review of 298 cases

Definitive identification of detached hooklets, scolex and fragments of the spiral wall of Cysticercus cellulosae on fine needle aspiration smears was documented. A critical review of aspirates from 200 diagnosed and 98 suspected lesions of soft tissue and intramuscular cysticercosis was done. Characteristic fragments of bladder wall cytomorphologically corresponding to viable or partially necrotic lesions were seen in 203 cases. Identification of fragments of an invaginated larva (i.e., hooklets, scolex or spiral wall) established the diagnosis in 33 of the suspected lesions. Cytomorphologically all these cases were from either necrotic or calcified lesions. In necrotic lesions with eosinophils, a careful search for fragments of the invaginated portion of the larva should be made.     

The use of trypsin and lysozyme immobilized on a cellulose carrier in treating otitis externa

In December 1991, an outbreak of Streptococcus bovis septicaemia occurred in a Belgian pigeon loft where 25 male and 25 female racing pigeons were housed. The main clinical signs included inability to fly and poor breeding results. None of the female pigeons and only one male pigeon was able to fly. Nine affected pigeons were necropsied. Histologically a tenosynovitis of the tendon of the deep pectoral muscle was observed in most of them and S bovis was isolated from the canalis triosseus or the shoulder joints of five of the nine pigeons. The pigeons were successfully treated with ampicillin administered in the drinking water for seven days.     

The pattern of residual muscle paralysis in poliomyelitis

Rural schoolchildren between the ages of 6 and 18 years were screened for residual paralysis following poliomyelitis; 503 (0.85%) out of a total of 58,592 were victims. Of these, only 16 (3.8%) had received poliomyelitis vaccine, while the remainder had not been immunised. The disease had occurred before the age of 4 years in nearly 90%, the lower limbs being affected (98%) more often than the upper (2%). In the lower limbs, the quadriceps, hip adductors and tibialis anterior were frequently affected. The muscles supplied by the L4 and L5 spinal segments were most commonly involved, while those supplied by L1 and S3 were least. Some order does exist in the apparently irregular distribution of paralysis after poliomyelitis.     

Non-medical drug use amongst non-student youth in India

In a survey of non-medical drug use by 266 non-students in the age range 10-24 years, tobacco, alcohol and cannabis were found to be the drugs more commonly used. Use was greater in males, older age groups, those with lower educational levels and those from urban areas. In comparison to students, among non-students use was practically limited to tobacco, alcohol and cannabis, the use was more regular and the age at onset lower. In a pilot trial of two additional strategies to identify drug users among non-student youth, namely cross-validation by fellow employees and verbal group administration of the questionnaire, the number of current users identified was very similar to that found by individual verbal administration, suggesting that these cheaper methods may be useful in some circumstances.     

Copper coating on carbon fibres and their composites with aluminium matrix

A uniform and continuous coating of copper was given to carbon fibres by cementation or electroless techniques. In both cases, when coating thicknesses were less than 0.2 m, copper deposition was discontinuous over the fibres, and above 0.2 m, coatings were continuous. In electroless coating, about 75% of the continuously coated fibres had a coating thickness range 0.2–0.5 m and above this showed isolated dendrite deposits of copper. In the cementation process, about 75% of the continuously coated fibres had a coating thickness range 0.2–0.6 m, and above this thickness, fine crystallite-type copper deposition was found over smoothly coated copper. The ultimate tensile strength of continuously electroless-coated fibres were nearer to the uncoated fibres, suggesting defect-free coating, while fibres coated by the cementation process exhibited lower ultimate tensile strength values. The tensile fracture of both electroless- and cementation-coated fibres showed delamination of the coating, suggesting poor bonding between coating and the fibre. In composites, prepared by dispersing the coated chopped fibres in a pure aluminium matrix, uniform and random distribution of the fibres were observed without appreciable fibre-metal interaction. The CuAl₂ intermetallics were largely found in the matrix and only very small amounts were observed at fibre/matrix interfaces. Additions of about 2 wt% Mg to the matrix prior to the fibre dispersion did not appreciably change the distribution pattern of the fibres, but in addition to CuAl₂ phase, Mg₂Si phases were observed in the matrix as well as at the interface.     

Phase I study of a five-day dose schedule of 4-Ipomeanol in patients with non-small cell lung cancer

The mammalian pulmonary toxin 4-ipomeanol (IPO) is activated by the cytochrome P450 system in bronchial Clara cells in animals. The resulting metabolites bind rapidly to macromolecules, producing localized cytotoxicity. IPO has in vitro and in vivo antitumor activity in non-small cell lung cancer (NSCLC) and thus was proposed as a lung cancer-specific antitumor agent. We have completed a directed Phase I trial in patients with NSCLC. Forty-four patients (34 men and 10 women) with NSCLC were treated with IPO. All but two patients had an Eastern Cooperative Oncology Group performance status of 0 or 1. They received 91 courses of therapy with i.v. IPO; 82 courses were administered daily for five days, and 9 were single bolus doses. The dose-limiting toxicity of elevated serum transaminases was observed in three of seven patients at 922 mg/m2/day. The maximum tolerated dose was 693 mg/m2/day on 5 consecutive days every 3 weeks. One patient developed grade 4 pulmonary toxicity at 167 mg/m2/day. There was no significant hematological or renal toxicity. No objective antitumor responses were observed. Pharmacokinetic analysis of 39 patients from day 1 of IPO administration showed biexponential elimination with mean half-lives of 8.6 (alpha half-life) and 76 min (beta half-life). There was a linear relationship between the area under the plasma drug concentration-time curve and the dose of IPO. There was no significant difference between the pharmacokinetic parameters measured on day 1 and day 5. Using a 4-day in vitro cytotoxicity assay, two tumor cell lines established from patients treated at 693 mg/m2/day had IC50s of approximately 6 mM, a concentration more than 75-fold higher than the plasma levels measured in these patients. Thus, although the total amount of drug administered per cycle on a daily times five dose schedule is more than 2.5-fold higher than the recommended single daily dose, IPO is unlikely to be a useful drug for patients with lung cancer.     

Disulfide bond mapping and structural characterization of spruce budworm antifreeze protein

We have previously reported that bryostation 1 (Bryo 1) induces differentiation of chronic lymphocytic leukemia (CLL) in vitro to a hairy cell (HC) stage. This study tests the hypothesis that Bryo 1-differentiated CLL cells are more susceptible to 2-chlorodeoxyadenosine (2-CdA) than parent CLL cells. A recently established EBV-negative CLL line (WSU-CLL) from a patient resistant to chemotherapy including fludarabine was used to test this hypothesis. Both Bryo 1 (10-1000 nM) and 2-CdA (5.6-22.4 microM) exhibited a dose-dependent growth inhibitory effect on the WSU-CLL cell line. In vitro, the sequential exposure to Bryo 1 (100 nM for 72 h) followed by 2-CdA (11.2 microM) resulted in significantly higher rates of growth inhibition than either agent alone. Changes in immunophenotype, enzymes, lipids, proteins, and the DNA of WSU-CLL cells were studied before and after Bryo 1 treatment. Bryo 1 induced a positive tartrate-resistant acid phosphatase reaction and two important markers, CD11c and CD25, after 72 h of culture, confirming the differentiation of CLL to HC. The Fourier transformation infrared spectroscopic analysis showed that the amount of membrane lipids significantly increased in Bryo 1-treated cells compared to controls after 24 h, whereas the protein content, as well as the DNA content, decreased. This finding supports the change of CLL to HC. To evaluate the in vivo efficacy of Bryo 1 and 2-CdA, we used a xenograft model of CLL in WSU-CLL-bearing mice with severe combined immune deficiency. s.c. tumors were developed by injection of 10(7) WSU-CLL cells, and fragments were then transplanted into a new batch of severe combined immunodeficient mice. Bryo 1 and 2-CdA at the maximum tolerated doses (75 micrograms/kg i.p. and 30 mg/kg s.c., respectively) were administered to the mice at different combinations and schedules. The survival in days, the tumor growth inhibition ratio, the tumor growth delay, and the log10 kill of the mice treated with Bryo 1 followed by 2-CdA were significantly better than the control and other groups. We conclude that the sequential treatment with Bryo 1 followed by 2-CdA resulted in higher antitumor activity and improved animal survival.