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M Y Hsu

Publications and source records attributed to M Y Hsu.

43 records · Page 3Linked to original sources

Further studies on the development of Wuchereria bancrofti in laboratory animals.

In an attempt to obtain a small animal laboratory model for Wuchereria bancrofti Mongolian gerbils and hamsters were infected with third-stage larvae of strains of the parasite from Indonesia and China. Gerbils were necropsied at 7 to 591 days and larvae recovered up to 219 days. The third molt occurred on or before 10 days and worms found after 14 days were in the fourth stage. More worms were recovered early in the infection (7 to 30 days) than in older infections. The worms were usually recovered from the testes, pelt and carcass and heart and lungs. An adult male worm was recovered on one occasion from the lung of a gerbil at 92 days. The parasite was also found to develop in hamsters with results similar to those in gerbils. Gerbils and hamsters were given antithymocyte sera before and after infection and more worms were recovered and the worms larger in size in some treated animals. Continued studies with geographic strains of W. bancrofti in various animals should be encouraged in order to find a small laboratory animal model for the parasite.

Animals↗

Preliminary observations on the development of larval filariae in Toxorhynchites species.

Brugia malayi and B. Pahangi microfilariae from gerbil intraperitoneal infections were inoculated into the thorax of male and female Toxorhynchites amboinesis and developed into third-stage larvae as early as 11 days. In a comparative study with Aedes togoi fed on microfilaremic gerbils, third-stage larvae were found at 10 days. Some third-stage larvae of B. malayi inoculated into gerbils developed to advanced stages. Third-stage larvae of Wuchereria bancrofti were recovered in low numbers from Tx. amboinesis and Tx. aurifluus inoculated with microfilariae recovered from human blood by membrane filtration. Development of all filarial species was similar in both male and female mosquitoes. Toxorhynchites species are plant feeders and therefore reduce the hazards of laboratory transmission of pathogenic agents. Because of their large size, manipulations with this mosquito species are easy and the size allows for a larger inoculum to be used. This group of mosquitoes should develop into useful laboratory vectors for the transmission of arthropod-borne diseases.

Animals↗

Improved separation of creatine kinase isoenzymes by use of DEAE-Sepharose Cl-6B.

We describe here a simple, rapid chromatographic procedure for quantitatively separating serum creatine kinase isoenzymes (EC 2.7.3.2), with diethylaminoethyl (DEAE)-Sepharose CL-6B as the anion-exchanger. We established the column bed height and the elution parameters by use of a simplex procedure. DEAE-Sepharose CL-6B equilibrated in tris(hydroxymethyl)aminomethane (50 mmol/L, pH 7.5, and containing 30 mmol of NaCl per liter) is packed into a miniature polystyrene column with bed dimensions of 0.7 x 1.5 cm. The DEAE-Sepharose column system was evaluated and compared with a DEAE-Sephadex A-50 column system. The results indicate that the Sepharose column yields MM, MB, and BB isoenzymes uniquely, without cross contamination. Coefficients of variation (CV's) for 10 replicate column assays were 2.8, 5.9, and 15.2% for 569 U of MM per liter, 82.3 U of MB per liter, and 9.0 U of BB per liter, respectively. The serum sample was enriched with baboon heart extract. Day-to-day reproducibility for a serum control assayed on 10 days yielded CV's of 4.6, 9.9, and 40.3% for 391, 45.3 and 1.9 U of isoenzymes MM, MB, and BB per liter, respectively. A reference interval for each isoenzymes was derived from data on 81 men and 63 women.

Animals↗

Experimental transmission of Wuchereria bancrofti to monkeys.

Infective larvae of Wuchereria bancrofti from laboratory-raised Culex pipiens fatigans and Aedes togoi mosquitoes fed on human volunteers in Jakarta, Indonesia (J strain) and Kinmen Island, China (K strain) were introduced into Taiwan monkeys (Macaca cyclopis) by subcutaneous inoculation, by foot puncture, or by permitting infected mosquitoes to feed weekly on the monkeys. Some animals were splenectomized and others were treated with varying regimens of immunosuppressants. Necropsy was done on monkeys that died or were killed and the entire bodies were examined for worms. A total of 78 monkeys (43 males and 35 females) were exposed to infection and parasites were found in 29% of the females and 63% of males. In infections of 38 days or less worms were recovered from the testes of males and the pelt, carcass and lymph nodes of both sexes, but after 42 days of infection most worms were in the testes of males, and a few were recovered from lymph nodes and carcasses of females. Worms recovered at 8-11 days were third-stage, those found between 14 and 38 days fourth-stage, and ones found between 42 and 103 days were young adults. After 148 days most were adults and microfilariae were seen in the uteri of female worms at 160 days and later. The parasites continued to grow in size with time. Microfilariae were detected in the blood of nine monkeys between 8 and 18 months and the patent period varied from 5-21 months. Microfilarial densities were low and erratic, and periodicity could not be determined. The effectiveness of methods of administering infections and the value of various treatment regimens seem uncertain; monkey antilymphocytic sera, however, appeared to have some influence. Parasites were found in 36% of the Taiwan monkeys given the J strain and 54% of those given the K strain. A limited number of M. mulatta (3), M.irus (fascicularis) (3) and Aotus trivirgatus (4) were also given infective larvae and adult W. bancrofti were recovered from the testes of one male M. mulatta and one male M. irus; uterine microfilariae were found in one female worm from the latter monkey. A. trivirgatus were negative. Low numbers of infective larvae recovered from mosquitoes fed on patent monkeys were introduced intermittently into seven clean monkeys and one became microfilaremic between 11 and 17 months postinoculation.

Aedes↗

Separation of five isoenzymes of serum lactate dehydrogenase by discontinuous gradient elution from a miniature ion-exchange column.

We describe a simple, fast chromatographic technique for quantitatively separating the five isoenzymes of lactate dehydrogenase (LD; EC 1.1.1.27) in serum. A 250-microL serum sample is applied to a 6.0 x 0.7 cm column of QAE-Sephadex A-50 and eluted stepwise with five different buffers. The isoenzyme fractions, assayed by the method of Wroblewski and LaDue [Proc. Soc. Exp. Biol, Med. 90, 210 (1955)], are stable at room temperature for 24 h. For all five isoenzymes the average within-day coefficient of variation is 4.3%; the day-to-day CV for 20 days is 6.3%. Of some common potentially interfering substances tested, only sodium fluoride (238 mmol/L) was found to do so, by slowing the elution from the column and making the fractions turbid. The expected range in international (IUB) united and percent for each isoenzyme was determined from data on 73 men and 70 women. From these data we calculate, by a percentile estimate of a nongaussian distribution, a normal range of LD-1/LD-2 ratio of 0.55--0.87 for men and 0.52--0.91 for women. The LD isoenzyme patterns in both normal and above-normal samples of 147 sera, as evaluated by the present column method and by electrophoresis, correlated well.

Brain↗

Heparin/endothelial cell growth supplement regulates matrix gene expression and prolongs life span of vascular smooth muscle cells through modulation of interleukin-1.

Vascular smooth muscle cells produce and respond to interleukin-1, a cytokine which modifies inflammation-associated vascular activities including the synthesis of extracellular matrix proteins. We have established vascular smooth muscle cells culture conditions in which heparin, in the presence of endothelial cell growth supplement, promotes cell proliferation and inhibits interleukin-1 and matrix protein expression. To test whether interleukin-1 mediates growth and matrix modulation by heparin/endothelial cell growth supplement, vascular smooth muscle cells were transfected with an Epstein-Barr virus-derived expression vector designed to express interleukin-1 antisense transcripts. RNase protection and ELISA assays demonstrated a complete block of interleukin-1 transcription and protein synthesis. Northern blot analysis also showed that interleukin-1 antisense decreased the expression of matrix genes such as type I collagen, fibronectin, and decorin similar to downregulation after heparin/endothelial cell growth supplement treatment. In contrast, the expression of versican was not affected, indicating a selective suppression of matrix proteins. In addition, interleukin-1 antisense significantly prolonged the life span of vascular smooth muscle cells in culture. Our data suggest that heparin/endothelial cell growth supplement induces matrix remodeling and controls growth and senescence of vascular smooth muscle cells through down-regulation of interleukin-1.

Adult↗