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Contains fulltext : 291657.pdf (Publisher’s version ) (Open Access) BACKGROUND: Anti-TNF drugs, such as infliximab, are associated with attenuated antibody responses after SARS-CoV-2 vaccination. We aimed to determine how the anti-TNF drug infliximab and the anti-integrin drug vedolizumab affect vaccine-induced neutralising antibodies against highly transmissible omicron (B.1.1.529) BA.1, and BA.4 and BA.5 (hereafter BA.4/5) SARS-CoV-2 variants, which possess the ability to evade host immunity and, together with emerging sublineages, are now the dominating variants causing current waves of infection. METHODS: CLARITY IBD is a prospective, multicentre, observational cohort study investigating the effect of infliximab and vedolizumab on SARS-CoV-2 infection and vaccination in patients with inflammatory bowel disease (IBD). Patients aged 5 years and older with a diagnosis of IBD and being treated with infliximab or vedolizumab for 6 weeks or longer were recruited from infusion units at 92 hospitals in the UK. In this analysis, we included participants who had received uninterrupted biological therapy since recruitment and without a previous SARS-CoV-2 infection. The primary outcome was neutralising antibody responses against SARS-CoV-2 wild-type and omicron subvariants BA.1 and BA.4/5 after three doses of SARS-CoV-2 vaccine. We constructed Cox proportional hazards models to investigate the risk of breakthrough infection in relation to neutralising antibody titres. The study is registered with the ISRCTN registry, ISRCTN45176516, and is closed to accrual. FINDINGS: Between Sept 22 and Dec 23, 2020, 7224 patients with IBD were recruited to the CLARITY IBD study, of whom 1288 had no previous SARS-CoV-2 infection after three doses of SARS-CoV-2 vaccine and were established on either infliximab (n=871) or vedolizumab (n=417) and included in this study (median age was 46·1 years [IQR 33·6-58·2], 610 [47·4%] were female, 671 [52·1%] were male, 1209 [93·9%] were White, and 46 [3·6%] were Asian). After three doses of SARS-CoV-2 vaccine, 50% neutralising titres (NT50s) were significantly lower in patients treated with infliximab than in those treated with vedolizumab, against wild-type (geometric mean 2062 [95% CI 1720-2473] vs 3440 [2939-4026]; p

We have identified GpsA, a predicted glycerol-3-phosphate dehydrogenase, as a virulence factor in the Lyme disease spirochete Borrelia (Borreliella) burgdorferi: GpsA is essential for murine infection and crucial for persistence of the spirochete in the tick. B. burgdorferi has a limited biosynthetic and metabolic capacity; the linchpin connecting central carbohydrate and lipid metabolism is at the interconversion of glycerol-3-phosphate and dihydroxyacetone phosphate, catalyzed by GpsA and another glycerol-3-phosphate dehydrogenase, GlpD. Using a broad metabolomics approach, we found that GpsA serves as a dominant regulator of NADH and glycerol-3-phosphate levels in vitro, metabolic intermediates that reflect the cellular redox potential and serve as a precursor for lipid and lipoprotein biosynthesis, respectively. Additionally, GpsA was required for survival under nutrient stress, regulated overall reductase activity and controlled B. burgdorferi morphology in vitro. Furthermore, during in vitro nutrient stress, both glycerol and N-acetylglucosamine were bactericidal to B. burgdorferi in a GlpD-dependent manner. This study is also the first to identify a suppressor mutation in B. burgdorferi: a glpD deletion restored the wild-type phenotype to the pleiotropic gpsA mutant, including murine infectivity by needle inoculation at high doses, survival under nutrient stress, morphological changes and the metabolic imbalance of NADH and glycerol-3-phosphate. These results illustrate how basic metabolic functions that are dispensable for in vitro growth can be essential for in vivo infectivity of B. burgdorferi and may serve as attractive therapeutic targets.

The conserved oligomeric Golgi (COG) complex, a multisubunit tethering complex of the CATCHR (complexes associated with tethering containing helical rods) family, controls membrane trafficking and ensures Golgi homeostasis by orchestrating retrograde vesicle targeting within the Golgi. In humans, COG defects lead to severe multisystemic diseases known as COG‐congenital disorders of glycosylation (COG‐CDG). The COG complex both physically and functionally interacts with all classes of molecules maintaining intra‐Golgi trafficking, namely SNAREs, SNARE‐interacting proteins, Rabs, coiled‐coil tethers, and vesicular coats. Here, we review our current knowledge of COG‐related trafficking and glycosylation defects in humans and model organisms, and analyze possible scenarios for the molecular mechanism of the COG orchestrated vesicle targeting.

In pathogenicity assessment, RNA-based analyses are important for the correct classification of variants, and require gene-specific cut-offs for allelic representation and alternative/aberrant splicing. Beside this, the diagnostic yield of RNA-based techniques capable to detect aberrant splicing or allelic loss due to intronic/regulatory variants has to be elaborated. We established a cDNA analysis for full-length transcripts (FLT) of the four DNA mismatch repair (MMR) genes to investigate the splicing pattern and transcript integrity with active/inhibited nonsense-mediated mRNA-decay (NMD). Validation was based on results from normal controls, samples with premature termination codons (PTC), samples with splice-site defects (SSD), and samples with pathogenic putative missense variants. The method was applied to patients with variants of uncertain significance (VUS) or unexplained immunohistochemical MMR deficiency. We categorized the allelic representation into biallelic (50 ± 10%) or allelic loss (≤10%), and >10% and 10% and 30% of VUS. Our method allows a standardized, systematic cDNA analysis of the MMR FLTs to assess the pathogenicity mechanism of VUS on RNA level, which will gain relevance for precision medicine and gene therapy. Diagnostic accuracy will be enhanced by detecting MMR defects in hitherto unsolved patients. The data generated will help to calibrate a high-throughput NGS-based mRNA-analysis and optimize prediction programs.

TPS12143 Background: Cancer-related cognitive impairment (CRCI) can include persistent memory symptoms, and affects many cancer survivors. Memory and Attention Adaptation Training (MAAT) is an evidence-based cognitive behavioral therapy (CBT) that improves CRCI with demonstrated efficacy in telehealth delivery. MAAT consists of 8 weekly (45-minute) video visits. The aims of this study are to confirm MAAT telehealth efficacy in a phase III RCT (MAAT versus Supportive Therapy; ST) across large catchment areas of two comprehensive cancer centers. A secondary aim is to evaluate treatment-induced brain activation as assessed by functional MRI (fMRI) in a subset of participants. We present remote treatment and data capture methods of this open NCI-sponsored (R01CA244673) randomized clinical trial (NCT 04586530). These methods have high success in participant accrual despite COVID-19 pandemic conditions, and can be readily adopted to other clinical trials to enhance rural/underserved enrollment. Methods: We are enrolling 200 adult, stage I-III breast cancer survivors 1-5 years post-chemotherapy with cognitive complaints. Individuals with CNS disease, previous brain injury, dementia or psychiatric disorder are excluded. All study procedures are completed from the participant’s home (except fMRI). Eligibility screening is a semi-structured phone interview followed by detailed informed consent online (Research Electronic Data Capture: REDCap) with staff phone guidance. Consented participants complete baseline brief phone-based neurocognitive assessment and validated patient-reported outcome measures (PROs) of cognition and quality of life via REDCap. Participants are randomized to MAAT or ST and assigned treating clinicians at respective cancer centers. All 8 visits are completed through secure telehealth platforms, followed by repeat phone/online assessment post-treatment and again at 6 months. Enrollment began in 3/2021. As of 1/2022 (9 months), 56 participants are enrolled (28% of the planned sample), 47 randomized (MAAT 24; ST 23), with 24 completing post-treatment assessments. If all assessments and treatment visits were in person, travel burden per participant is 968 miles/20.5 hours driven, and $542 (US 2021 Federal rate). Thus, study travel savings to date are $30,352. Participant feedback indicates telehealth makes participation possible, similar to previous MAAT research. The current RCT demonstrates utility, efficiency and cost-savings of telehealth and remote data capture technology in the conduct of cancer control research. Elements of methods described can also be adopted for cancer therapeutic trials. Comprehensive cancer centers, where most clinical trials are based, can enhance participation of remote and/or underserved populations that have higher rates of cancer, more disease burden and less opportunity for trial participation. Clinical trial information: NCT04586530.

BACKGROUND: In Thailand, there are few pediatric emergency medicine (PEM) fellowship‐trained providers, and emergency departments (EDs) are staffed by pediatricians and emergency physicians. Our pediatric ED collaborated with Thailand's largest private hospital system to develop a training program designed to improve emergency care for children. OBJECTIVE: The objective was to develop, implement, and assess the efficacy of a curriculum to improve PEM care by emergency providers in a Thai health system. METHODS: We conducted a prospective study from January 2018 to July 2019. After an initial needs assessment, we developed a curriculum with 22 modules, divided into basic and advanced courses. Each course began with baseline testing, consisting of written tests and several simulated cases led by physician–nurse teams. Each course was administered longitudinally through electronic modules over 6 months, with in‐person shadowing and skills workshops at the course midpoint, culminating in final summative examinations and repeat simulation testing. RESULTS: On written examination scores for the basic course, physicians (n = 5) improved by 37.6% (95% confidence interval [CI] = 29.8% to 45.4%, p

Although orthopoxviruses (OPXV) are known to encode a majority of the genes required for replication in host cells, genome-wide genetic screens have revealed that several host pathways are indispensable for OPXV infection. Through a haploid genetic screen, we previously identified several host genes required for monkeypox virus (MPXV) infection, including the individual genes that form the conserved oligomeric Golgi (COG) complex. The COG complex is an eight-protein (COG1&ndash, COG8) vesicle tethering complex important for regulating membrane trafficking, glycosylation enzymes, and maintaining Golgi structure. In this study, we investigated the role of the COG complex in OPXV infection using cell lines with individual COG gene knockout (KO) mutations. COG KO cells infected with MPXV and vaccinia virus (VACV) produced small plaques and a lower virus yield compared to wild type (WT) cells. In cells where the KO phenotype was reversed using a rescue plasmid, the size of virus plaques increased demonstrating a direct link between the decrease in viral spread and the KO of COG genes. KO cells infected with VACV displayed lower levels of viral fusion and entry compared to WT suggesting that the COG complex is important for early events in OPXV infection. Additionally, fewer actin tails were observed in VACV-infected KO cells compared to WT. Since COG complex proteins are required for cellular trafficking of glycosylated membrane proteins, the disruption of this process due to lack of individual COG complex proteins may potentially impair the virus-cell interactions required for viral entry and egress. These data validate that the COG complex previously identified in our genetic screens plays a role in OPXV infection.

COG is a multisubunit vesicle tethering complex in the Golgi. We demonstrate that both COG subcomplexes can be permanently attached to Golgi membranes and that major COG functions do not require cycling between the membrane and cytosol., The conserved oligomeric Golgi (COG) complex is a vesicle tether of the “complexes associated with tethering containing helical rods” family, which functions on the cytoplasmic side of Golgi. It is currently unknown whether COG function, or function of any multisubunit vesicular tether, depends on cycling between the membrane and cytosol. Therefore, we permanently anchored key subunits of COG subcomplexes (COG4, COG7, and COG8) to Golgi membranes using transmembrane protein TMEM115 (TMEM-COG). All TMEM-COG subunits tested were Golgi localized, integrated into the COG complex, and stabilized membrane association of endogenous subunits. Interestingly, TMEM-COG4 and TMEM-COG7 equally rescued COG function in organization of Golgi markers, glycosylation, and abundance of COG-sensitive proteins. In contrast, TMEM-COG8 was not as effective, indicating that N-terminal attachment of COG8 interfered with overall COG structure and function, and none of the TMEM-COG subunits rescued the abnormal Golgi architecture caused by COG knockout. Collectively, these data indicate that both subcomplexes of the COG complex can perform most of COG function when permanently attached to membranes and that the cytosolic pool of COG is not completely essential to COG function.

The conserved oligomeric complex (COG) is a multi-subunit vesicle tethering complex that functions in retrograde trafficking at the Golgi. We have previously demonstrated that the formation of enlarged endo-lysosomal structures (EELSs) is one of the major glycosylation-independent phenotypes of cells depleted for individual COG complex subunits. Here, we characterize the EELSs in HEK293T cells using microscopy and biochemical approaches. Our analysis revealed that the EELSs are highly acidic and that vATPase-dependent acidification is essential for the maintenance of this enlarged compartment. The EELSs are accessible to both trans-Golgi enzymes and endocytic cargo. Moreover, the EELSs specifically accumulate endolysosomal proteins Lamp2, CD63, Rab7, Rab9, Rab39, Vamp7, and STX8 on their surface. The EELSs are distinct from lysosomes and do not accumulate active Cathepsin B. Retention using selective hooks (RUSH) experiments revealed that biosynthetic cargo mCherry-Lamp1 reaches the EELSs much faster as compared to both receptor-mediated and soluble endocytic cargo, indicating TGN origin of the EELSs. In support to this hypothesis, EELSs are enriched with TGN specific lipid PI4P. Additionally, analysis of COG4/VPS54 double KO cells revealed that the activity of the GARP tethering complex is necessary for EELSs’ accumulation, indicating that protein mistargeting and the imbalance of Golgi-endosome membrane flow leads to the formation of EELSs in COG-deficient cells. The EELSs are likely to serve as a degradative storage hybrid organelle for mistargeted Golgi enzymes and underglycosylated glycoconjugates. To our knowledge this is the first report of the formation of an enlarged hybrid endosomal compartment in a response to malfunction of the intra-Golgi trafficking machinery.

A nine-year-old castrated male miniature dachshund presented for chronic pustular dermatitis, lethargy, fever and weight loss. There was no history of dermatological disease prior to lesion development which occurred within days following administration of afoxolaner (NexGard) and worsened with each of the five subsequent doses. Physical examination revealed multifocal alopecia, diffuse crusting and peripheral lymphadenomegaly. Multiple intact panfollicular pustules were present in the inguinal region, concave pinnae and ear canals. Cytology from intact pustules revealed non-degenerate neutrophils, acantholytic keratinocytes with no bacterial or fungal organisms. Deep skin scrape and dermatophyte culture were negative. Full-thickness cutaneous punch biopsy samples of intact pustules were obtained, and histopathological findings included subcorneal acantholytic pustular dermatosis. This was consistent with the clinical findings and the diagnosis of pemphigus foliaceus. Treatment was initiated with immunosuppressive prednisone and cyclosporine and remission was achieved. Medications were slowly tapered, though low doses were required to maintain long-term remission.

A 4-year-old American Quarter Horse gelding was evaluated for acute non–weight-bearing lameness of the right thoracic limb with swelling in the right shoulder region. Physical examination revealed radial nerve paralysis of unknown etiology. The primary differential diagnosis was musculoskeletal trauma. Ultrasonography of the right shoulder region identified a heterogeneous mass that extended from the point of the shoulder to the thoracic inlet. Cytologic analysis of fluid collected by fine needle aspirate of the mass was consistent with large cell lymphoma. Based on the cytological findings, locally invasive neoplasia was diagnosed and considered the likely cause of the radial nerve paralysis. Because of the poor prognosis, the horse was euthanized, and postmortem examination confirmed the diagnosis of a nonclassified large cell lymphoma that extended from the deep tissues of the right pectoral muscle group into the thoracic inlet and pleural cavity, as well as the right brachial plexus. The mass in the region of the brachial plexus encompassed and mechanically compressed all of the nerves within the area, resulting in the clinical sign of radial nerve paralysis. Although neoplasia as a cause of radial nerve paralysis is rare, it should be considered as a differential diagnosis, regardless of age.

Background Although intermediate alleles (27–35 CAG repeats) do not fall within the range to cause Huntington’s Disease (HD), they are susceptible to paternal germline instability, and so may expand into the reduced penetrance or full mutation range upon transmission to the next generation. The risk of IAs expanding into the HD disease causing range has been difficult to establish. Aims and methods At the EHDN conference in 2016, we presented two cases, where low risk intermediate alleles unexpectedly expanded into the full mutation range. Having discussed these cases locally, we had differing opinions whether to offer predictive testing to family members and whether prenatal testing or PGD would be offered. We recommended the need to develop a consensus guideline for the counselling of IAs between different genetic centres throughout the EHDN. In order to study this, we developed an online survey to determine practice between centres. This survey uses the two case examples we presented previously, to ascertain how different genetics health professionals would counsel each family regarding the risk of the IA expanding, and the implications for family members. Results and conclusions To date, we have received 25 survey responses from genetic counsellors, geneticists and HD researchers working in different centres worldwide. We present the results of this questionnaire which highlight the ongoing challenges associated with genetic counselling for IAs and the difference in practice between centres. The survey identifies a need for consensus guidelines, with over 60% of participants expressing a need for further guidance in this area.

The conserved oligomeric Golgi (COG) complex controls membrane trafficking and ensures Golgi homeostasis by orchestrating retrograde vesicle trafficking within the Golgi. Human COG defects lead to severe multisystemic diseases known as COG-congenital disorders of glycosylation (COG-CDG). To gain better understanding of COG-CDGs, we compared COG knockout cells with cells deficient to 2 key enzymes, Alpha-1,3-mannosyl-glycoprotein 2-beta-N-acetylglucosaminyltransferase and uridine diphosphate-glucose 4-epimerase (GALE), which contribute to proper N- and O-glycosylation. While all knockout cells share similar defects in glycosylation, these defects only account for a small fraction of observed COG knockout phenotypes. Glycosylation deficiencies were not associated with the fragmented Golgi, abnormal endolysosomes, defective sorting and secretion or delayed retrograde trafficking, indicating that these phenotypes are probably not due to hypoglycosylation, but to other specific interactions or roles of the COG complex. Importantly, these COG deficiency specific phenotypes were also apparent in COG7-CDG patient fibroblasts, proving the human disease relevance of our CRISPR knockout findings. The knowledge gained from this study has important implications, both for understanding the physiological role of COG complex in Golgi homeostasis in eukaryotic cells, and for better understanding human diseases associated with COG/Golgi impairment.

We examine psychometric properties and validity of the Perceptions of Attraction scale (POA). Responses of undergraduate students (N = 510) indicate that the POA has high reliability and a coherent factor structure comprising two dimensions of feelings of attraction toward others and perceptions of attraction from others that explain 67% of response variance. POA scores are correlated with social skills and extraversion, but not related to other personality dimensions. POA scores are predictive of outcomes in current romantic relationships (n = 160).

Many intracellular pathogens exploit host secretory trafficking to support their intracellular cycle, but knowledge of these pathogenic processes is limited. The bacterium Brucella abortus uses a Type IV secretion system (VirB T4SS) to generate a replication-permissive Brucella-containing vacuole (rBCV) derived from the host endoplasmic reticulum (ER), a process that requires host early secretory trafficking. Here we show that the VirB T4SS effector BspB contributes to rBCV biogenesis and Brucella replication by interacting with the Conserved Oligomeric Golgi (COG) tethering complex, a major coordinator of Golgi vesicular trafficking, thus remodeling Golgi membrane traffic and redirecting Golgi-derived vesicles to the BCV. Altogether, these findings demonstrate that Brucella modulates COG-dependent trafficking via delivery of a T4SS effector to promote rBCV biogenesis and intracellular proliferation, providing mechanistic insight into how bacterial exploitation of host secretory functions promotes pathogenesis.

Background Our program's most recent ACGME Resident Survey revealed that residents felt inadequately prepared to perform many required procedures without supervision. Moreover, current residents noted a poor understanding about which procedures are considered “essential” by the ACGME and independently voiced a desire for more dedicated training in emergent stabilization of critically ill children. Objective Improve procedural competency in pediatric residents through education, dedicated procedural time, and simulation. Methods Our program pursued the following methods: 1) Design and distribute a procedure passport for resident reference. This credit card sized tool delineates the 13 “essential” ACGME procedures and includes a target number of procedures to complete on certain rotations. 2) Develop a half day experience where residents work with a nurse completing procedures such as intravenous catheter placement, venipuncture, and bladder catheterization in the ED. 3) Organize intensive small-group sessions to emphasize bag valve mask ventilation and pediatric intubation. Each resident completes these half day simulated code experiences annually in addition to our monthly mock code curriculum. Results We obtained survey responses from residents about the utilization of the passport, and 47% felt it helped them better understand ACGME requirements. Additionally, 24% felt the passport helped them take initiative to complete procedures independently. Of the residents paired with nurses, 92% felt it was valuable. In addition to the previously listed procedures, residents also completed arterial draws, intraosseous lines, incisions and drainage, laceration repairs, and nasogastric tube placements. Conclusions A clear and tangible delineation of ACGME requirements helps educate and support residents independently completing procedures. Furthermore, increasing clinical time in areas with high procedural volume seems to be both well regarded and clinically beneficial. The expanded use of simulation has promise as an educational tool for procedural practice, and we are currently evaluating its utility as well as launching a procedural elective.

PUF family translational repressors are conserved developmental regulators, but the molecular function provided by the regions flanking the PUF RNA-binding domain is unknown. In C. elegans, the PUF proteins FBF-1 and FBF-2 support germline progenitor maintenance by repressing production of meiotic proteins and use distinct mechanisms to repress their target mRNAs. We identify dynein light chain DLC-1 as an important regulator of FBF-2 function. DLC-1 directly binds to FBF-2 outside of the RNA-binding domain and promotes FBF-2 localization and function. By contrast, DLC-1 does not interact with FBF-1 and does not contribute to FBF-1 activity. Surprisingly, we find that the contribution of DLC-1 to FBF-2 activity is independent of the dynein motor. Our findings suggest that PUF protein localization and activity are mediated by sequences flanking the RNA-binding domain that bind specific molecular partners. Furthermore, these results identify a new role for DLC-1 in post-transcriptional regulation of gene expression.

The conserved oligomeric Golgi (COG) complex is a peripheral membrane protein complex which orchestrates tethering of intra-Golgi vesicles. We found that COG1-4 (lobe A) and 5–8 (lobe B) protein assemblies are present as independent sub-complexes on cell membranes. Super-resolution microscopy demonstrates that COG sub-complexes are spatially separated on the Golgi with lobe A preferential localization on Golgi stacks and the presence of lobe B on vesicle-like structures, where it physically interacts with v-SNARE GS15. The localization and specific interaction of the COG sub-complexes with the components of vesicle tethering/fusion machinery suggests their different roles in the vesicle tethering cycle. We propose and test a novel model that employs association/disassociation of COG sub-complexes as a mechanism that directs vesicle tethering at Golgi membranes. We demonstrate that defective COG assembly or restriction of tethering complex disassembly by a covalent COG1-COG8 linkage is inhibitory to COG complex activity, supporting the model.

The Conserved Oligomeric Golgi complex is an evolutionarily conserved multisubunit tethering complex (MTC) that is crucial for intracellular membrane trafficking and Golgi homeostasis. The COG complex interacts with core vesicle docking and fusion machinery at the Golgi; however, its exact mechanism of action is still an enigma. Previous studies of COG complex were limited to the use of CDGII (Congenital disorders of glycosylation type II)-COG patient fibroblasts, siRNA mediated knockdowns, or protein relocalization approaches. In this study we have used the CRISPR approach to generate HEK293T knock-out (KO) cell lines missing individual COG subunits. These cell lines were characterized for glycosylation and trafficking defects, cell proliferation rates, stability of COG subunits, localization of Golgi markers, changes in Golgi structure, and N-glycan profiling. We found that all KO cell lines were uniformly deficient in cis/medial-Golgi glycosylation and each had nearly abolished binding of Cholera toxin. In addition, all cell lines showed defects in Golgi morphology, retrograde trafficking and sorting, sialylation and fucosylation, but severities varied according to the affected subunit. Lobe A and Cog6 subunit KOs displayed a more severely distorted Golgi structure, while Cog2, 3, 4, 5, and 7 knock outs had the most hypo glycosylated form of Lamp2. These results led us to conclude that every subunit is essential for COG complex function in Golgi trafficking, though to varying extents. We believe that this study and further analyses of these cells will help further elucidate the roles of individual COG subunits and bring a greater understanding to the class of MTCs as a whole.

The conserved oligomeric Golgi (COG) complex is a key evolutionally conserved multisubunit protein machinery that regulates tethering and fusion of intra-Golgi transport vesicles. The Golgi apparatus specifically promotes sorting and complex glycosylation of glycoconjugates. Without proper glycosylation and processing, proteins and lipids will be mislocalized and/or have impaired function. The Golgi glycosylation machinery is kept in homeostasis by a careful balance of anterograde and retrograde trafficking to ensure proper localization of the glycosylation enzymes and their substrates. This balance, like other steps of membrane trafficking, is maintained by vesicle trafficking machinery that includes COPI vesicular coat proteins, SNAREs, Rabs, and both coiled-coil and multi-subunit vesicular tethers. The COG complex interacts with other membrane trafficking components and is essential for proper localization of Golgi glycosylation machinery. Here we describe using CRISPR-mediated gene editing coupled with a phenotype-based selection strategy directly linked to the COG complex's role in glycosylation homeostasis to obtain COG complex subunit knockouts (KOs). This has resulted in clonal KOs for each COG subunit in HEK293T cells and gives the ability to further probe the role of the COG complex in Golgi homeostasis.

The study purpose was to identify practitioner competencies occupational therapists perceive as important for handwriting evaluation and intervention in school-age children. Examination of the influence of practice setting and years of experience were also investigated. A national sample of 376 occupational therapists participated via survey. A total of 68 of 80 items were perceived to be of high importance. The top five identified items were eye-hand coordination, motor planning, review of work samples/direct observation, visual motor integration, and provision of education to teachers, parents and administrators. Analysis indicated minimal difference due to practice setting in the domains of handwriting and assessment of legibility. No significant differences based on practitioner experience were revealed. Eighty-five percent of the items were perceived as highly important. Practitioner perceptions of competency were mainly similar regardless of practice setting or experience.

Background Dermatophytic pseudomycetoma is an atypical form of dermatophytosis where the infection is located in the deep dermal and subcutaneous tissues. Although rare, it is most commonly associated with Microsporum canis or Trichophyton sp. It has been reported in cats, dogs and horses. Objectives To describe the clinical and pathological findings of dermatophytic pseudomycetoma caused by M. canis and Trichophyton sp. in two domestic ferrets. Animals Two pet ferrets (Mustela putorius furo). Method and materials Case report. Results Two ferrets were diagnosed with dermatophytic pseudomycetoma confirmed by histological examination of tissue and fungal culture. In both cases, ferrets presented with several cutaneous firm nodules 0.6-3 cm in diameter. Microscopic lesions revealed multifocal nodular pyogranulomatous inflammation with intralesional fungi. Conclusions and clinical importance To the best of the authors' knowledge, this is the first description of dermatophytic pseudomycetoma in domestic ferrets. This disease should be included in the differential diagnosis of nodular dermatopathies in this species.

Cooperative storage involves two or more libraries sharing space within a facility. Kansas State University leases space at the Library Annex, a remote storage facility owned and operated by the University of Kansas. Various challenges are involved in maintaining access to remotely stored materials for resource sharing and document delivery purposes. The authors, staff members of the Interlibrary Services Unit at Kansas State University, discuss variables affecting access to remote materials and share their experience working within a cooperative storage model.

This article examines the evolving role of HIM professionals in research. In terms of both growth and credibility, it is important that HIM as a profession become more integrally involved in the research process. Specifically, this article examines the expanding role of the HIM professional in research-related activities and data management activities. For illustrative purposes, examples from research in the areas of medication error detection, utilization of web technologies in QI processes, and creation of an HIT infrastructure to connect healthcare providers in which HIM faculty members are actively involved in collaborative research projects with pharmacy, nursing, and medicine faculty in an academic medical center setting are provided.

A description of 7 consecutive patients admitted for various forms of neurological impairment. All these patients were found to have cerebrospinal fluid positive for West Nile virus IgM, and routine bacterial cultures were negative. Six of these patients did not present with the typical lymphocytic pleocytosis often quoted when discussing a viral meningitis/encephalitis; rather most presented with a cerebrospinal fluid neutrophilia.

In recent years, microscopy systems have become more capable of capturing, transferring, and manipulating electronic images. In the last issue of this publication, there was a summary of different image format types and their flexibility for use with microscopy. This article will briefly discuss converting images among different formats, which often becomes necessary when transferring images from one software application to another or creating hardcopy of the image. In order to transfer the image, either the original software has to save the image to a format which the new software or output device can read, or that new software or device has to read a format which is native to the image generating software. Sometimes (happy days), this all works out straightforwardly. Other times, transferring an image is impossible. The user then has the choice of writing his own conversion software, finding freeware, or purchasing a commercial conversion utility.

Because the growing and exporting of fresh bananas is one of the few comparative advantages of some Latin American countries, it is imparative that the maximum revenue is generated from these operations. The procedures for setting prices through the channels of distribution and the price elasticity of demand at the retail level are estimated from market information acquired in the study. With the application of an equal export tax, the exporting countries could then realize the increased revenues from a small price increase. By using marketing research techniques rather than the traditional econometric methods, the authors develop a strategic approach that should increase the export earnings of the banana producing countries by as much as $73-80 million annually.