Developing New Online Course Accessibility Services for Faculty through Collaboration between Librarians and Campus Departments: A Case Study.

Literature has shown that inaccessible content is a barrier to student success and an impediment to student retention. Despite legal obligations for accessible course content, creators of course materials are often unaware of the benefits of improved accessibility and their personal liability. To address these accessibility issues, a partnership was developed between the library and two campus departments to create a formal, campus wide accessibility service that would make all online course content fully accessible on Day 1, through design initiatives rather than having faculty wait for an accommodation request, to foster student success and support faculty course development.

The effects of metabolic syndrome, obesity, and the gut microbiome on load-induced osteoarthritis.

OBJECTIVE: Metabolic syndrome is characterized by obesity, hyperglycemia, hypertension, insulin resistance, and dyslipidemia. Metabolic syndrome is associated with osteoarthritis (OA), but it is unclear if the association is attributable to increased mechanical loading on joints caused by obesity or other aspects of metabolic syndrome. Here we examined the effects of altered metabolism, obesity, and the gut microbiome on load-induced OA. DESIGN: Cartilage damage was induced through cyclic compressive loading in four groups of adult male mice: Toll-like receptor-5 deficient (TLR5KO) mice that develop metabolic syndrome due to alterations in the gut microbiome, TLR5KO mice submitted to chronic antibiotics to prevent metabolic syndrome (TLR5KOΔMicrobiota), C57BL/6J mice fed a high fat diet to cause obesity (HFD), and untreated C57BL/6J mice (WT). Loading was applied for 2 weeks (n = 10-11/group) or 6 weeks (n = 10-11/group). RESULTS: After 2 weeks of loading, cartilage damage (OARSI score) was not different among groups. After 6 weeks of loading, HFD mice had increased load-induced cartilage damage, while TLR5KO mice had cartilage damage comparable to WT mice. TLR5KOΔMicrobiota mice had less cartilage damage than other groups. HFD mice had elevated serum inflammatory markers. Each group had a distinct gut microbiome composition. CONCLUSIONS: Severe obesity increased load-induced cartilage damage, while milder changes in adiposity/metabolic syndrome seen in TLR5KO mice did not. Furthermore, the effects of systemic inflammation/obesity on cartilage damage depend on the duration of mechanical loading. Lastly, reduced cartilage damage in the TLR5KOΔMicrobiota mice suggests that the gut microbiome may influence cartilage pathology.

Recurrent hyperactive ESR1 fusion proteins in endocrine therapy-resistant breast cancer.

Background: Estrogen receptor-positive (ER-positive) metastatic breast cancer is often intractable due to endocrine therapy resistance. Although ESR1 promoter switching events have been associated with endocrine-therapy resistance, recurrent ESR1 fusion proteins have yet to be identified in advanced breast cancer. Patients and methods: To identify genomic structural rearrangements (REs) including gene fusions in acquired resistance, we undertook a multimodal sequencing effort in three breast cancer patient cohorts: (i) mate-pair and/or RNAseq in 6 patient-matched primary-metastatic tumors and 51 metastases, (ii) high coverage (>500×) comprehensive genomic profiling of 287-395 cancer-related genes across 9542 solid tumors (5216 from metastatic disease), and (iii) ultra-high coverage (>5000×) genomic profiling of 62 cancer-related genes in 254 ctDNA samples. In addition to traditional gene fusion detection methods (i.e. discordant reads, split reads), ESR1 REs were detected from targeted sequencing data by applying a novel algorithm (copyshift) that identifies major copy number shifts at rearrangement hotspots. Results: We identify 88 ESR1 REs across 83 unique patients with direct confirmation of 9 ESR1 fusion proteins (including 2 via immunoblot). ESR1 REs are highly enriched in ER-positive, metastatic disease and co-occur with known ESR1 missense alterations, suggestive of polyclonal resistance. Importantly, all fusions result from a breakpoint in or near ESR1 intron 6 and therefore lack an intact ligand binding domain (LBD). In vitro characterization of three fusions reveals ligand-independence and hyperactivity dependent upon the 3' partner gene. Our lower-bound estimate of ESR1 fusions is at least 1% of metastatic solid breast cancers, the prevalence in ctDNA is at least 10× enriched. We postulate this enrichment may represent secondary resistance to more aggressive endocrine therapies applied to patients with ESR1 LBD missense alterations. Conclusions: Collectively, these data indicate that N-terminal ESR1 fusions involving exons 6-7 are a recurrent driver of endocrine therapy resistance and are impervious to ER-targeted therapies.

Metabolomic identification of diagnostic serum-based biomarkers for advanced stage melanoma.

INTRODUCTION: Melanoma is a highly aggressive malignancy and is currently one of the fastest growing cancers worldwide. While early stage (I and II) disease is highly curable with excellent prognosis, mortality rates rise dramatically after distant spread. We sought to identify differences in the metabolome of melanoma patients to further elucidate the pathophysiology of melanoma and identify potential biomarkers to aid in earlier detection of recurrence. METHODS: Using 1H NMR and DI-LC-MS/MS, we profiled serum samples from 26 patients with stage III (nodal metastasis) or stage IV (distant metastasis) melanoma and compared their biochemical profiles with 46 age- and gender-matched controls. RESULTS: We accurately quantified 181 metabolites in serum using a combination of 1H NMR and DI-LC-MS/MS. We observed significant separation between cases and controls in the PLS-DA scores plot (permutation test p-value = 0.002). Using the concentrations of PC-aa-C40:3, DL-carnitine, octanoyl-L-carnitine, ethanol, and methylmalonyl-L-carnitine we developed a diagnostic algorithm with an AUC (95% CI) = 0.822 (0.665-0.979) with sensitivity and specificity of 100 and 56%, respectively. Furthermore, we identified arginine, proline, tryptophan, glutamine, glutamate, glutathione and ornithine metabolism to be significantly perturbed due to disease (p < 0.05). CONCLUSION: Targeted metabolomic analysis demonstrated significant differences in metabolic profiles of advanced stage (III and IV) melanoma patients as compared to controls. These differences may represent a potential avenue for the development of multi-marker serum-based assays for earlier detection of recurrences, allow for newer, more effective targeted therapy when tumor burden is less, and further elucidate the pathophysiologic changes that occur in melanoma.

Zoonotic Thelazia californiensis in dogs from New Mexico, USA, and a review of North American cases in animals and humans.

Thelazia californiensis is a spirurid nematode found in the conjunctival sac of domestic and wild animals, including humans, across the western United States of America. Herein, we report two cases of thelaziosis by T. californiensis in dogs from New Mexico, United States, based on integrated morphological and molecular approaches. Nematode specimens collected from the conjunctiva of both dogs were identified as T. californiensis based on morphology. Our study substantially expands the knowledge on morphometry of this nematode species. Therefore, these data will be useful for accurate diagnosis of thelaziosis in domestic animals, wildlife and humans in North America, using classical, microscopy-based methods. We characterized for the first time the mitochondrial cytochrome c-oxidase subunit 1 (COI) and 12S genes of T. californiensis. While these markers support the validity of T. californiensis, they were not very informative for elucidating the phylogenetic relationships among Thelazia species. Nevertheless, the characterization of these diagnostic markers for T. californiensis will be useful for studies on the epidemiology, molecular xenomonitoring of fanniid vectors, and population genetics of this multi-host, zoonotic parasite.

Sparganosis due to Spirometra sp. (cestoda; Diphyllobothriidae) in captive meerkats (Suricata suricatta).

We report three cases of sparganosis due to plerocercoids of the tapeworm Spirometra sp. in captive meerkats (Suricata suricatta) from a zoo exhibit in the southeastern United States. Two meerkats were euthanized, one due to an uncontrollable seizure and the other due to trauma, and at necropsy cysts containing cestode larvae were observed. A third meerkat had a subcutaneous nodule surgically removed, which contained similar larvae. The third animal died years later, and had numerous cestode larvae in the pleural and peritoneal cavities. The larvae were morphologically identified as plerocercoids of diphyllobothriidean cestodes. On necropsy, multiple nodules, ranging in size from 2.5 to 3.0 cm, were observed in the subcutaneous tissue and muscles. Multifocally, separating skeletal muscle fibers were longitudinal and transversal sections of cestode larva. Histologically, parasitic cysts contained large numbers of neutrophils and macrophages, admixed with proteinaceous material. Molecular and phylogenetic analyses confirmed that specimens from one of the meerkats belonged to the genus Spirometra and was closely related to Spirometra plerocercoids isolated from a snake from the United States and wild felids from South America. Meerkats likely became infected by ingesting infected second intermediate hosts, such as amphibians and reptiles that may have entered the exhibit. Management practices that minimize access of meerkats and other susceptible hosts to intermediate hosts should be implemented.

Tryptophan operon: structural gene mutation creating a "promoter" and leading to 5-methyltryptophan dependence.

A strain carrying the mutation trpA515, which maps in the "unusual" region (between the genes trpA and trpB) of the tryptophan operon of Salmonella typhimurium is capable of utilizing anthranilic acid as a growth factor only in the presence of the analog 5-methyltryptophan, normally a potent growth inhibitor. The reason for this peculiar phenotype is the creation by trpA515 of a transcription-initiating signal in the "unusual" region.

Reduction to homozygosity of genes on chromosome 11 in human breast neoplasia.

The somatic loss of heterozygosity for normal alleles occurring in human tumors has suggested the presence of recessive oncogenes. The results presented here demonstrate a loss of heterozygosity of several genes on chromosome 11 in primary breast tumors. Restriction fragment length polymorphism analysis of these DNAs further suggests that the most frequent loss of sequences in breast tumors occurs between the beta-globin and parathyroid hormone loci on the short arm of chromosome 11. The loss of heterozygosity for chromosome 11 loci has a significant association with tumors that lack estrogen and progesterone receptors, grade III tumors, and distal metastasis.

Major pol gene progenitors in the evolution of oncoviruses.

The genetic relationships among molecularly cloned prototype viruses representing all of the major oncovirus genera were investigated by molecular hybridization and nucleotide sequence analysis. One of the major progenitors of the pol genes of such viruses gives rise to mammalian type C viruses and another gives rise to type A, B, D, and avian type C oncoviruses. Evidence of unusual patterns of homology among the env genes of mammalian type C and D oncoviruses illustrates that genetic interactions between their progenitors contributed to the evolution of oncoviruses.

Bovine leukemia virus genes in the DNA of leukemic cattle.

Reverse transcripts of the rna genome of the bovine leukemia virus (BLV) as well as 125I-labeled BLV RNA hybridize to the DNA of tissues from leukemic cattle with the adult form of the disease but not to bovine thymic lymphoma or normal bovine tissues.

A new class of endogenous human retroviral genomes.

Human DNA contains multiple copies of a novel class of endogenous retroviral genomes. Analysis of a human recombinant DNA clone (HLM-2) containing one such proviral genome revealed that it is a mosaic of retroviral-related sequences with the organization and length of known endogenous retroviral genomes. The HLM-2 long terminal repeat hybridized with the long terminal repeat of the squirrel monkey virus, a type D retrovirus. The HLM-2 gag and pol genes share extensive nucleotide sequence homology with those of the M432 retrovirus (a type A-related retrovirus), mouse mammary tumor virus (a type B retrovirus), and the avian Rous sarcoma virus (a type C retrovirus). Nucleotide sequence analysis revealed regions in the HLM-2 pol gene that were as much as 70 percent identical to the mouse mammary tumor virus pol gene. A portion of the putative HLM-2 env gene hybridized with the corresponding region of the M432 viral genome.

Randomized trial to evaluate contraceptive efficacy, safety and acceptability of a two-rod contraceptive implant over 4 years in the Dominican Republic.

OBJECTIVE: Sino-implant (II) is a contraceptive implant that had a commodity price one-third of the competing products a decade ago. To make Sino-implant (II) more widely available, we conducted a trial to collect safety and efficacy data required for World Health Organization (WHO) prequalification, a quality standard allowing global donors to procure a pharmaceutical product. STUDY DESIGN: This was a randomized controlled trial allocating 650 participants to either Sino-implant (II) or Jadelle®. Participants were seen at 1 and 6 months, and then semiannually. The primary efficacy measure was the pregnancy Pearl Index [number of pregnancies per 100 women-years (WY) of follow-up] in the Sino-implant (II) group during up to 4 years of implant use. RESULTS: For the primary outcome, Sino-implant (II) had a 4-year Pearl Index of 0.74 (95% confidence interval, 0.36-1.37) compared to 0.00 (95% confidence interval, 0.00-1.04) for Jadelle®. The Sino-implant (II) pregnancy rate was significantly higher in the fourth year (3.54 per 100 WY) than in the first 3 years combined (0.18 per 100 WY; p <.001). Total levonorgestrel concentrations were equivalent between groups at month 12, but were 19%, 22% and 32% lower in the Sino-implant (II) group at months 24, 36 and 48, respectively (p <.001 at each time point). Safety and acceptability of the two products were similar, while providers documented significantly higher breakage rates during removal of Sino-implant (II) (16.3% vs. 3.1%; p <.001). CONCLUSION: Based on these results, WHO prequalified Sino-Implant (II) with a 3-year use label in June 2017, 2 years shorter than the 5-year duration of Jadelle®. IMPLICATIONS: WHO prequalification allows global donors to procure Sino-implant (II), which means women in many low resource countries will have greater access to highly effective and acceptable contraceptive implants. Our study noted important clinical differences, including shorter duration of high effectiveness with Sino-implant (II) when compared to the other available two-rod system, Jadelle®. Introduction strategies should include appropriate training on these differences.

Kit and PDGFR-alpha activities are necessary for Notch4/Int3-induced tumorigenesis.

Transgenic mice overexpressing Notch4 intracellular domain (Int3) under the control of the whey acidic protein (WAP) or mouse mammary tumor virus-long terminal repeat promoters, develop mammary tumors. Microarray analysis of these tumors revealed high levels of c-Kit expression. Gleevec is a tyrosine kinase inhibitor that targets c-Kit, platelet-derived growth factor receptors (PDGFRs) and c-Abl. This led us to speculate that tyrosine kinase receptor activity might be a driving force in the development of Int3 mammary tumors. WAP-Int3 tumor-bearing mice were treated with continuous release of Gleevec using subcutaneously implanted Alzet pumps. Phosphorylation of c-Kit, PDGFRs and c-Abl is inhibited in Int3 transgenic mammary tumors by Gleevec. Inhibition of these enzymes is associated with a decrease in cell proliferation and angiogenesis, and an induction of apoptosis. To examine the signaling mechanisms underlying Notch4/Int3 tumorigenesis, we employed small interfering RNA (siRNA) to knock down c-Kit, PDGFRs and c-Abl alone or in combination and observed the effects on soft agar growth of HC11 cells overexpressing Int3. Only siRNA constructs for c-Kit and/or PDGFR-alpha were able to inhibit HC11-Int3 colony formation in soft agar. Our data demonstrate an inhibitory effect of Gleevec on Int3-induced transformation of HC11 cells and mammary tumors and indicate an oncogenic role for c-Kit and PDGFR-alpha tyrosine kinases in the context of Int3 signaling.

A cryptic species of Onchocerca (Nematoda: Onchocercidae) in blackflies (Simulium spp.) from southern California, USA.

BACKGROUND: Entomological surveillance for pathogens based on molecular screening of putative arthropod vectors such as blackflies (Diptera: Simuliidae) is becoming increasingly important. Surveillance provides a means to understand host and geographical patterns of underestimated biodiversity among North American species of Onchocerca and a pathway to identify and track expanding emergence of the zoonotic Onchocerca lupi. Herein, we have screened two blackfly species, Simulium tescorum and Simulium vittatum (s.l.), from Los Angeles County, southern California, USA for DNA of filarioid nematodes to better understand species richness and limits within the genus Onchocerca. METHODS: A total of 1056 and 378 female blackflies was collected using CO2-baited mosquito traps from March to November of 2015 and 2016, respectively. All blackflies during 2015 were individually processed for DNA extraction and PCR targeting of the cytochrome c oxidase subunit 1 (cox1) of the mitochondrial DNA (mtDNA). Specimens of S. tescorum collected in 2016 were processed individually with heads and bodies extracted separately, whereas those of S. vittatum (s.l.) were processed in pooled samples with heads and bodies extracted separately. A subset of filarioid-positive samples from 2015 and all samples from 2016 were screened using a PCR targeting the NADH dehydrogenase subunit 5 (nad5) gene (mtDNA). RESULTS: In 2015, 356 S. tescorum (33.7%) and 683 S. vittatum (s.l.) (64.7%) were collected, and an additional 17 specimens were not assessed morphologically. In 2016, a total of 378 blackflies was collected. Of these, 43 (11.6%) were S. tescorum and 327 (88.4%) were S. vittatum (s.l.), and an additional 8 specimens were not assessed morphologically. In 2015, Onchocerca sequences were detected in 4.8% (n = 17) of S. tescorum samples, and only one S. vittatum (0.15%). In 2016, only a single S. vittatum pool was positive for the same cryptic Onchocerca species. In phylogenetic comparisons based on nad5, the Onchocerca sequences from California formed a clade with those isolates in white-tailed deer from upstate New York, suggesting these belong to a single widespread cryptic species. CONCLUSIONS: An uncharacterized species of Onchocerca associated with cervid hosts was found in blackflies from southern California. Sequence data demonstrated it is likely conspecific with an unnamed species of Onchocerca previously found in white-tailed deer from upstate New York. Current data support recognition of a broad geographical distribution across North America for an apparently cryptic species of Onchocerca that is discrete from O. cervipedis, considered to be a typical filarioid among cervids. Our data suggest that this cryptic species of Onchocerca may infect subspecies of white-tailed deer (Odocoileus virginianus), and mule and black-tailed deer (Odocoileus hemionus) at temporal latitudes. The blackflies Simulium tescorum and S. vittatum (s.l.) (presumably, S. tribulatum) are putative vectors. Discovery of a cryptic complex indicates that species diversity and putative associations for definitive hosts and vectors of Onchocerca species in North America must be reassessed.

Targeting risk factors for type 2 diabetes in American Indian youth: the Tribal Turning Point pilot study.

BACKGROUND: American Indian (AI) youth are at high risk for type 2 diabetes. OBJECTIVES: To partner with Eastern Band of Cherokee Indians and Navajo Nation to develop a culturally sensitive behavioural intervention for youth (Tribal Turning Point; TTP) and assess feasibility in an 8-month randomized pilot study. METHODS: We enrolled 62 overweight/obese AI children (7-10 years) who participated with ≥1 parent/primary caregiver. Intervention participants (n = 29) attended 12 group classes and five individual sessions. Control participants (n = 33) attended three health and safety group sessions. We analysed group differences for changes in anthropometrics (BMI, BMI z-score, waist circumference), cardiometabolic (insulin, glucose, blood pressure) and behavioural (physical activity and dietary self-efficacy) outcomes. RESULTS: Study retention was 97%, and intervention group attendance averaged 84%. We observed significant treatment effects (p = 0.02) for BMI and BMI z-score: BMI increased in control (+1.0 kg m-2 , p < 0.001) but not intervention participants (+0.3 kg m-2 , p = 0.13); BMI z-score decreased in intervention (-0.17, p = 0.004) but not control participants (0.01, p = 0.82). There were no treatment effects for cardiometabolic or behavioural outcomes. CONCLUSIONS: We demonstrated that a behavioural intervention is feasible to deliver and improved obesity measures in AI youth. Future work should evaluate TTP for effectiveness, sustainability and long-term impact in expanded tribal settings.

Mutations in human breast cancer: an overview.

Studies of mammary tumorigenesis in mice infected with the mouse mammary tumor virus and in certain strains of transgenic mice with an activated oncogene have provided strong evidence that multiple mutations contribute to the initiation and progression of malignancies in the breast. The increasing availability of recombinant DNA probes that detect various proto-oncogenes, growth factor genes, and growth factor receptor genes, as well as restriction fragment length polymorphisms in the human population, have made possible a molecular approach for the identification of frequently occurring mutations in primary human breast tumor DNA. The aim of studies using this molecular approach has been to investigate whether specific mutations are highly associated with various clinical parameters, including disease prognosis. Eight mutations have been identified, including amplification of c-myc, c-erbB2, and int-2, as well as loss of heterozygosity on five chromosomes (1q, 3p, 11p, 13q, and 17p). Loss of heterozygosity is thought to unmask recessive mutations of tumor-suppressor genes. In some studies, amplification of either c-myc, c-erbB2, or int-2 has been found to have a significant association with high risk of relapse or poor survival. The current status of these mutations as potentially useful prognostic indicators for the management of the disease is controversial and points to the need for further research in this area.

Presence of two members of c-erbA receptor gene family (c-erbA beta and c-erbA2) in smallest region of somatic homozygosity on chromosome 3p21-p25 in human breast carcinoma.

The loss of heterozygosity of genes on the short arm of chromosome 3 (3p) in human breast carcinomas occurs in a region involved in other malignancies, including renal cell carcinoma, lung cancers, and von Hippel-Lindau disease. This finding suggests the presence of a gene(s) that plays a crucial role in multiple cancers. In our study of 84 informative (heterozygous) primary breast tumors, 30% showed losses of heterozygosity on chromosome 3. The shortest region of homozygosity in primary human breast tumor is located between the DNF15S2 and RAF1 loci in the 3p21-p25 region on the short arm of chromosome 3. This region includes at least two members of the c-erbA steroid/thyroid hormone receptor family (c-erbA beta and c-erbA2) that may be of special relevance to breast cancer. Furthermore, tumors with a loss of heterozygosity of genes on chromosome 3 were previously reported to have frequent allelic deletions on chromosome 11p and amplification of the c-myc proto-oncogene. These results highlight the occurrence of multiple genetic alterations in breast tumors.

Detection of novel murine mammary tumor viruses by interspecies immunoassays.

Radioimmunoassays were developed that can detect antigenic determinants common to mammary tumor viruses (MTV's) of four distinct Mus species: M. musculus, M. cervicolor, M. cookll, and M. caroll. The radioimmunoassays were based on the immunologic cross-reactivity observed between the murine mammary tumor viruses (MuMTV) of M. musculus and type B retrovirus isolated from M. cervicolor. Both of the glycoproteins of MuMTV (gp52, gp36) shared antigenic determinants with virions of M. cervicolor mammary tumor virus. Interspecies radioimmunoassays for gp52 and gp36 were developed and used to detect viruses in the milk of noninbred feral Mus species and MuMTV-related translational products in mammary tumors in these species. Type C and type D retroviruses, as well as the M432 retrovirus of M. cervicolor, did not react in either assay. Both interspecies immunoassays were therefore specific for the detection of distinct MuMTV-related antigenic determinants.