Huh WK et al. (2003) Nature "Global analysis of protein localization in budding yeast."

Huh WK, Howson RW, Weissman JS, Gerke LC, Falvo JV, Carroll AS, Desbordes SC

[Nature, 2003]

A fundamental goal of cell biology is to define the functions of proteins in the context of compartments that organize them in the cellular environment. Here we describe the construction and analysis of a collection of yeast strains expressing full-length, chromosomally tagged green fluorescent protein fusion proteins. We classify these proteins, representing 75% of the yeast proteome, into 22 distinct subcellular localization categories, and provide localization information for 70% of previously unlocalized proteins. Analysis of this high-resolution, high-coverage localization data set in the context of transcriptional, genetic, and protein-protein interaction data helps reveal the logic of transcriptional co-regulation, and provides a comprehensive view of interactions within and between organelles in eukaryotic cells.

Uchikawa R et al. (1991) J Parasitol "Antibody responses and protective immunity in rats receiving repeated inoculations of Strongyloides ratti."

Ichiki H, Uchikawa R, Komaki E

[J Parasitol, 1991]

The relationship between specific antibody responses and protective immunity against Strongyloides ratti was examined in rats receiving 10, 50, or 500 infective larvae (L3) at weekly intervals. No specific IgG response was detected in rats receiving 10-L3 inoculations for 7 wk. Fifty- and 500-L3 inoculations induced an IgG response by weeks 2 and 3, respectively, and a higher IgG response was induced in rats receiving the higher doses. All 3 inoculation doses induced high IgE responses, but the kinetics were different. IgE in the 10-L3 group continued to rise from weeks 4 to 7. In the 50- and 500-L3 groups, IgE was detected first at week 3 and increased until week 5. It then declined in the 500-L3 group and the titer at week 7 was significantly lower than that at week 5, whereas it remained the same in the 50-L3 group. The number of larvae recovered from the head 40 hr after a challenge inoculation (1,000 L3) significantly declined by weeks 7, 3, and 2 in rats receiving 10, 50-, and 500-L3 inoculations, respectively. Intestinal worm burdens increased for 7 wk in the 10-L3 group, 5 wk for the 50-L3 group, and 2 wk for the 500-L3 group. These findings indicate that repeated inoculations of low doses of L3 induce delayed and limited protective immunity to a heavy challenge and worm expulsion from the intestine. There was a temporal correlation between the levels of protection and serum IgG, whereas circulating IgE level did not seem to affect directly either the level of the resistance or expulsion of intestinal worms.

Suzuki N et al. (1999) Biol Reprod "Expression of trophinin, tastin, and bystin by trophoblast and endometrial cells in human placenta."

Nakayama J, Aoki D, Shih IM, Nozawa S, Suzuki N, Fukuda MN

[Biol Reprod, 1999]

Trophinin, tastin, and bystin comprise a complex mediating a unique homophilic cell adhesion between trophoblast and endometrial epithelial cells at their respective apical cell surfaces. In this study, we prepared mouse monoclonal antibodies specific to each of these molecules. The expression of these molecules in the human placenta was examined immunohistochemically using the antibodies. In placenta from the 6th week of pregnancy, trophinin and bystin were found in the cytoplasm of the syncytiotrophoblast in the chorionic villi, and in endometrial decidual cells at the utero placental interface. Tastin was exclusively present on the apical side of the syncytiotrophoblast. Tissue sections were also examined by in situ hybridization using RNA probes specific to each of these molecules. This analysis showed that trophoblast and endometrial epithelial cells at the utero placental interface express trophinin, tastin, and bystin. In wk 10 placenta, trophinin and bystin were found in the intravillous cytotrophoblast, while tastin was not found in the villi. After wk 10, levels of all three proteins decreased and then disappeared from placental villi.

Rajan TV et al. (1992) J Parasitol "Survival of Onchocerca volvulus in nodules implanted in immunodeficient rodents."

Nelson FK, Cupp E, Greiner DL, Rajan TV, Schultz LD

[J Parasitol, 1992]

Onchocerca volvulus is an obligate human parasite, and its study has been difficult due to an inability to maintain it outside the human host. We report the successful transplantation of onchocercomata containing living adult O. volvulus worms into immunodeficient C.B.-17.scid/scid (scid) mice or athymic rnu/rnu (nude) rats. Living, motile worms containing viable microfilariae were present in onchocercomata recovered from scid mice or nude rats for up to 20 wk, establishing a novel animal model for future investigation of O. volvulus.

Uchikawa R et al. (1989) J Parasitol "The effects of single and repeated inoculations of various larval doses on Strongyloides ratti burden and distribution in rats."

Uchikawa R, Sato A, Nojima H

[J Parasitol, 1989]

The changes in worm burden, distribution, length, and fecundity after and during single and repeated inoculations of 10, 50, or 500 larvae of Strongyloides ratti were examined in rats. Worm burden after a single inoculation of a higher larval dose reduced rapidly. Repeated inoculations of lower larval doses at weekly intervals led to a delayed peak and slower reduction of worm burden; the repeated inoculations of 10 larvae did not induce worm expulsion for at least 7 wk. In repeated inoculations at 3-wk intervals, a primary inoculation of 500 larvae induced strong resistance to reinfection at week 3, whereas no resistance was induced until week 6 in rats receiving repeated inoculations of 10 or 50 larvae. Similar dose-dependent reductions in worm length and fecundity were observed in single and repeated inoculations, and the reductions began earlier than worm expulsion. Intestinal migration of worms from the upper small intestine to the large intestine was observed during the course of single and repeated inoculations. Earlier and clearer migration was observed in rats receiving higher doses. These findings indicate that in S. ratti infection, the changes of worm burden, distribution, length, and fecundity are dependent on the inoculated larval dose.

Fong LY et al. (2000) Proc Natl Acad Sci U S A "Muir-Torre-like syndrome in Fhit-deficient mice."

Fong LY, Fidanza V, Ottey M, Lock LF, Siprashvili Z, McCue PA, Siracusa LD, Huebner K, Mancini R, Zanesi N, Druck T, Martin SE, Croce CM

[Proc Natl Acad Sci U S A, 2000]

To investigate the role of the Fhit gene in carcinogen induction of neoplasia, we have inactivated one Fhit allele in mouse embryonic stem cells and produced (129/SvJ x C57BL/6J) F(1) mice with a Fhit allele inactivated (+/-). Fhit +/+ and +/- mice were treated intragastrically with nitrosomethylbenzylamine and observed for 10 wk posttreatment. A total of 25% of the +/+ mice developed adenoma or papilloma of the forestomach, whereas 100% of the +/- mice developed multiple tumors that were a mixture of adenomas, squamous papillomas, invasive carcinomas of the forestomach, as well as tumors of sebaceous glands. The visceral and sebaceous tumors, which lacked Fhit protein, were similar to those characteristic of Muir-Torre familial cancer syndrome.

Rodriguez PH et al. (1999) J Am Mosq Control Assoc "Reduction in susceptibility to Brugia malayi of F2 progeny of Aedes togoi treated with ethyl methanesulfonate."

Wilson P, Benoit D, Castillon R, Rodriguez PH, Nasr-Schirf D

[J Am Mosq Control Assoc, 1999]

The susceptibility to Brugia malayi infection was tested in F2 female progeny derived from male and female Aedes togoi treated with ethyl methanesulfonate (EMS). Three-day-old males and females were treated with 0.025, 0.050, and 0.075, 0.10, 0.15, or 0.20% EMS by allowing them to feed for 5 days on sugar cubes containing EMS and then mated at random. Percentage of susceptibility and mean number of infective larvae (L3) in F2 females were analyzed over a 2-wk period. Reductions in susceptibility were significant in the F2 populations arising from the 3 highest EMS concentrations. F2 infections were reduced by 80%, indicating that EMS-induced mutations affect loci associated with filarial nematode susceptibility.

Joshi B et al. (2020) Evid Based Complement Alternat Med "Antibacterial, Antifungal, Antiviral, and Anthelmintic Activities of Medicinal Plants of Nepal Selected Based on Ethnobotanical Evidence."

Neyts J, Parajuli N, Jouneghani RS, Luyten W, Liu M, Joshi B, Panda SK, Leyssen P

[Evid Based Complement Alternat Med, 2020]

Background: Infections by microbes (viruses, bacteria, and fungi) and parasites can cause serious diseases in both humans and animals. Heavy use of antimicrobials has created selective pressure and caused resistance to currently available antibiotics, hence the need for finding new and better antibiotics. Natural products, especially from plants, are known for their medicinal properties, including antimicrobial and anthelmintic activities. Geoclimatic variation, together with diversity in ethnomedicinal traditions, has made the Himalayas of Nepal an invaluable repository of traditional medicinal plants. We studied antiviral, antibacterial, antifungal, and anthelmintic activities of medicinal plants, selected based upon ethnobotanical evidence. Methods: . Also, cytotoxicity was assessed on human hepatoma (Huh), rhabdosarcoma (RD), and Vero (VC) cell lines. Results: motility, comparable to levamisole. Conclusions: In countries like Nepal, with a high burden of infectious and parasitic diseases, and a current health system unable to combat the burden of diseases, evaluation of local plants as a treatment or potential source of drugs can help expand treatment options. Screening plants against a broad range of pathogens (bacteria, viruses, fungi, and parasites) will support bioprospecting in Nepal, which may eventually lead to new drug development.

Cox-Paulson EA et al. (2012) CBE Life Sci Educ "Studying human disease genes in Caenorhabditis elegans: a molecular genetics laboratory project."

Batzli JM, Grana TM, Cox-Paulson EA, Harris MA

[CBE Life Sci Educ, 2012]

Scientists routinely integrate information from various channels to explore topics under study. We designed a 4-wk undergraduate laboratory module that used a multifaceted approach to study a question in molecular genetics. Specifically, students investigated whether Caenorhabditis elegans can be a useful model system for studying genes associated with human disease. In a large-enrollment, sophomore-level laboratory course, groups of three to four students were assigned a gene associated with either breast cancer (brc-1), Wilson disease (cua-1), ovarian dysgenesis (fshr-1), or colon cancer (mlh-1). Students compared observable phenotypes of wild-type C. elegans and C. elegans with a homozygous deletion in the assigned gene. They confirmed the genetic deletion with nested polymerase chain reaction and performed a bioinformatics analysis to predict how the deletion would affect the encoded mRNA and protein. Students also performed RNA interference (RNAi) against their assigned gene and evaluated whether RNAi caused a phenotype similar to that of the genetic deletion. As a capstone activity, students prepared scientific posters in which they presented their data, evaluated whether C. elegans was a useful model system for studying their assigned genes, and proposed future directions. Assessment showed gains in understanding genotype versus phenotype, RNAi, common bioinformatics tools, and the utility of model organisms.

Miller JA et al. (2009) CBE Life Sci Educ "A laboratory-intensive course on RNA interference and model organisms."

Miller JA, Carson S, Witherow DS

[CBE Life Sci Educ, 2009]

RNA interference (RNAi) is a powerful method to silence gene expression in a variety of organisms and is generating interest not only as a useful tool for research scientists but also as a novel class of therapeutics in clinical trials. Here, we report that undergraduate and graduate students with a basic molecular biology background were able to demonstrate conceptual knowledge and technical skills for using RNAi as a research tool upon completion of an intensive 8-wk RNAi course with a 2-h lecture and 5-h laboratory per week. Students were instructed on design of RNAi experiments in model organisms and perform multiweek laboratory sessions based on journal articles read and discussed in class. Using Nicotiana benthamiana, Caenorhabditis elegans, and mammalian cell culture, students analyzed the extent of silencing using both qualitative assessment of phenotypic variations and quantitative measurements of RNA levels or protein levels. We evaluated the course over two semesters, each with a separate instructor. In both semesters, we show students met expected learning outcomes as demonstrated by successful laboratory experiment results, as well as positive instructor assessments of exams and lab reports. Student self-assessments revealed increased confidence in conceptual knowledge and practical skills. Our data also suggest that the course is adaptable to different instructors with varying expertise.