SDU - software for high-throughput automated data collection at the Swiss Light Source.

Recent advances in automation have fostered the development of unattended data collection services at a handful of synchrotron facilities worldwide. At the Swiss Light Source, the installation of new high-throughput sample changers at all three macromolecular crystallography beamlines and the commissioning of the Fast Fragment and Compound Screening pipeline created a unique opportunity to automate data acquisition. Here, the DA+ microservice software stack upgrades, implementation of an automatic loop-centering service and deployment of the Smart Digital User (SDU) software for unattended data collection are reported. The SDU software is the decision-making software responsible for communications between services, sample and device safety, sample centering, sample alignment with grid based X-ray diffraction and, finally, data collection.

Patients' Reactions to Letters Communicating Collateral Findings of Pragmatic Clinical Trials: a National Web-Based Survey.

BACKGROUND: Collateral findings in pragmatic clinical trials are findings that may have implications for patients' health but were not generated to address a trial's primary research questions. It is uncertain how best to communicate these collateral findings to patients. OBJECTIVES: To determine how reactions to a letter communicating collateral findings relate to who signed the letter, the type of finding, or whether the letter specified that the finding arose from a pragmatic clinical trial. RESEARCH DESIGN: Web-based survey experiment using a between-subjects design in which respondents were randomly assigned within education strata to view and respond to 1 of 16 hypothetical scenarios. SUBJECTS: Adults recruited from an online panel constructed from a probability sample of US-based postal addresses. MEASURES: The primary outcomes were the action the respondent would take next (i.e., contact a doctor immediately or something else) and the respondent's emotional reactions (i.e., all positive, all negative, mixed, or none). RESULTS: A total of 4080 respondents had analyzable data. Although some effects were statistically significant (P < .05), none exceeded a prespecified threshold for policy relevance (15 or more percentage points). Ratings of letter clarity and level of understanding were lower for letters that included a description of the clinical trial. CONCLUSIONS: Signatory and level of detail about collateral findings did not substantially affect people's intentions to take the recommended action of contacting their doctor. Deciding whether to include a description of the pragmatic clinical trial requires a trade-off between transparency and more difficulty understanding the contents of the letter.

Interaction specificity of clustered protocadherins inferred from sequence covariation and structural analysis.

Clustered protocadherins, a large family of paralogous proteins that play important roles in neuronal development, provide an important case study of interaction specificity in a large eukaryotic protein family. A mammalian genome has more than 50 clustered protocadherin isoforms, which have remarkable homophilic specificity for interactions between cellular surfaces. A large antiparallel dimer interface formed by the first 4 extracellular cadherin (EC) domains controls this interaction. To understand how specificity is achieved between the numerous paralogs, we used a combination of structural and computational approaches. Molecular dynamics simulations revealed that individual EC interactions are weak and undergo binding and unbinding events, but together they form a stable complex through polyvalency. Strongly evolutionarily coupled residue pairs interacted more frequently in our simulations, suggesting that sequence coevolution can inform the frequency of interaction and biochemical nature of a residue interaction. With these simulations and sequence coevolution, we generated a statistical model of interaction energy for the clustered protocadherin family that measures the contributions of all amino acid pairs at the interface. Our interaction energy model assesses specificity for all possible pairs of isoforms, recapitulating known pairings and predicting the effects of experimental changes in isoform specificity that are consistent with literature results. Our results show that sequence coevolution can be used to understand specificity determinants in a protein family and prioritize interface amino acid substitutions to reprogram specific protein-protein interactions.

Combining High-Throughput Synthesis and High-Throughput Protein Crystallography for Accelerated Hit Identification.

Protein crystallography (PX) is widely used to drive advanced stages of drug optimization or to discover medicinal chemistry starting points by fragment soaking. However, recent progress in PX could allow for a more integrated role into early drug discovery. Here, we demonstrate for the first time the interplay of high throughput synthesis and high throughput PX. We describe a practical multicomponent reaction approach to acrylamides and -esters from diverse building blocks suitable for mmol scale synthesis on 96-well format and on a high-throughput nanoscale format in a highly automated fashion. High-throughput PX of our libraries efficiently yielded potent covalent inhibitors of the main protease of the COVID-19 causing agent, SARS-CoV-2. Our results demonstrate, that the marriage of in situ HT synthesis of (covalent) libraires and HT PX has the potential to accelerate hit finding and to provide meaningful strategies for medicinal chemistry projects.

Patients' Views About the Disclosure of Collateral Findings in Pragmatic Clinical Trials: a Focus Group Study.

BACKGROUND: Pragmatic clinical trials (PCTs) are increasingly being conducted to efficiently generate evidence to inform healthcare decision-making. Despite their growing acceptance, PCTs may involve a variety of ethical issues, including the management of pragmatic clinical trial-collateral findings (PCT-CFs), that is, information that emerges in PCTs that is unrelated to the primary research questions but may have implications for patients, clinicians, and health systems. OBJECTIVE: We sought to understand patients' views about PCT-CF disclosure, including how, by whom, and the nature and extent of information provided. DESIGN: Prospective, qualitative focus group study. PARTICIPANTS: Focus groups were conducted in Baltimore, MD; Houston, TX; and Seattle, WA (overall N = 66), during July and August 2019. APPROACH: All groups discussed a hypothetical scenario involving the detection of a PCT-CF of contraindicated medications. Participants were asked about their reactions to the PCT-CF and issues related to its disclosure. KEY RESULTS: Reactions to learning about the PCT-CF were mixed, ranging from fear of a significant health problem, anger that the contraindicated medications had gone unnoticed and/or for being included in research without their permission, to gratitude for the information. Preferences for how such disclosures are made varied but were driven by several consistent desires, namely minimizing patient harm and anxiety and demonstrating trust and respect. Many wanted their treating clinician to be informed of the PCT-CF so that they would be prepared to answer patients' questions and to discuss treatment options. CONCLUSIONS: The detection of PCT-CFs is likely to increase with further expansion of PCTs. As such, clinicians will undoubtedly become involved in the management of PCT-CFs. Our data illustrate some of the challenges clinicians may face when their patients are informed of a PCT-CF and the need to develop guidance for disclosing PCT-CFs in ways that align with patients' preferences and values.

Feline bilateral inflammatory aural polyps: a descriptive retrospective study.

BACKGROUND: Feline inflammatory aural polyps (IP) have been reported as solitary growths, yet bilateral polyps may occur more commonly than described previously. OBJECTIVES: To identify the prevalence and risk factors associated with bilateral feline IP within a population of cats evaluated at a veterinary teaching hospital over a 10 year period. ANIMALS: Twenty-five cats with histologically confirmed IP. METHODS AND MATERIALS: A retrospective study was performed by searching computerized medical records of cats seen at The University of Tennessee Veterinary Teaching Hospital from 2005 to 2015. Keywords used for the search included "feline polyp", "ventral bulla osteotomy", "inflammatory polyp ear canal" and "nasopharyngeal polyp". RESULTS: Cats were separated into groups with unilateral or bilateral disease. Six of 25 (24%) cats had bilateral IP. Twelve cats had computed tomography performed (four of six with bilateral IP; eight of 19 with unilateral IP). Clinical signs, history of upper respiratory infection, post-treatment complications, and IP recurrence with and without local and systemic steroid administration were studied between groups and no differences were identified. In four of six cats with bilateral disease, the second polyp was identified with advanced imaging performed at the time of referral. CONCLUSIONS AND CLINICAL IMPORTANCE: These findings demonstrate that bilateral IP occur more frequently than reported previously and establish a prevalence for bilateral IP. These findings also highlight the value of advanced imaging techniques in diagnosing bilateral IP.

Antibacterial effect of N-acetylcysteine in combination with antimicrobials on common canine otitis externa bacterial isolates.

BACKGROUND: Approved treatments for canine otitis externa are limited in variety and may contain ototoxic ingredients. With bacterial resistance an ongoing concern, it would be ideal if non-ototoxic agents combined with antibiotics resulted in a synergistic effect, requiring lower antibiotic concentrations to treat infections. Evidence of synergism and antagonism between N-acetylcysteine (NAC) and various antibiotic classes has been reported; the present research group was interested in examining these interactions. HYPOTHESIS/OBJECTIVES: To determine if NAC, an otoprotective and antimicrobial compound, has synergistic activity when combined with enrofloxacin or gentamicin in vitro against bacterial isolates causing canine otitis externa. ANIMALS: Twenty-two isolates from canine clinical cases of otitis externa were identified and tested, including seven Staphylococcus pseudintermedius, 12 Pseudomonas aeruginosa and three Corynebacterium spp. isolates. METHODS AND MATERIALS: Each isolate was grown on blood agar for 24 h and transferred to Mueller-Hinton broth (MHB), with a final concentration of 5 × 105 cfu/mL. Each well was inoculated with 50 µL of bacterial suspension. N-acetylcysteine was diluted in MHB to a starting concentration of 160 mg/mL. Enrofloxacin and gentamicin were diluted to 64 µg/mL. Individual and checkerboard serial microdilution assays were performed in triplicate with negative controls for all isolates tested. RESULTS: Interactions observed for NAC and enrofloxacin were synergistic (4.5%), indifferent (77.3%) or antagonistic (18.2%). Interactions observed for NAC and gentamicin were synergistic (4.5%), indifferent (45.5%) or antagonistic (50%). CONCLUSIONS AND CLINICAL RELEVANCE: Most interactions between NAC and enrofloxacin or gentamicin were indifferent or antagonistic at the concentrations tested in vitro.

Description and characterization of a hair coat disorder in schipperkes.

BACKGROUND: Schipperkes develop bilaterally symmetrical alopecia preceded by lightening of the hair coat not associated with systemic clinical signs. The alopecia is presumptively diagnosed as Alopecia X but has never been investigated. HYPOTHESIS/OBJECTIVES: The purpose of this study was to describe clinical features, histopathological and laboratory abnormalities associated with symmetrical, noninflammatory alopecia in schipperkes. ANIMALS: Three healthy and eight affected schipperkes. METHODS AND MATERIALS: Complete blood count (CBC), serum chemistry panel, urinalysis (UA), urine cortisol: creatinine ratio (UCCR), total thyroxine (T4 ), free thyroxine by equilibrium dialysis (fT4 ed), thyroid stimulating hormone (TSH) and pre- and post-stimulation cortisol with sex hormone analyses were performed for all dogs. A minimum of two skin biopsy sites were sampled for histopathological evaluation. RESULTS: The CBC, serum chemistry panel, UA, T4 , fT4 ed, TSH and post-ACTH cortisol concentrations were normal for all dogs. Androstenedione concentrations were increased for all dogs; two of three healthy and five of eight affected dogs had increased post-ACTH androstenedione concentrations. Estradiol concentrations were increased for all healthy and four of eight affected dogs, with post-ACTH estradiol concentrations increased for all healthy and five of eight affected dogs. Progesterone concentrations were increased for two of three healthy and four of eight affected dogs. Post-ACTH progesterone concentrations were increased for one of three healthy and four of eight affected dogs. For one of three healthy and two of eight affected dogs, 17-hydroxyprogesterone concentrations were increased; post-ACTH 17-hydroxyprogesterone concentrations were increased in three of eight affected dogs only. Histopathology results from affected dogs were consistent with hair cycle arrest. CONCLUSIONS AND CLINICAL IMPORTANCE: This disorder resembles Alopecia X clinically and histologically.

The prevalence of Dermatophilus congolensis in horses with pastern dermatitis using PCR to diagnose infection in a population of horses in southern USA.

BACKGROUND: Dermatophilus congolensis is a facultative anaerobic actinomycete that causes papular to exudative dermatitis with crusting in horses. This organism is frequently implicated as a cause of pastern dermatitis, but few data are available validating the organism's association with this disease. HYPOTHESIS/OBJECTIVES: The aim of this study was to evaluate if D. congolensis is associated with pastern dermatitis in horses utilizing RT-qPCR. ANIMALS: Fifteen client-owned horses diagnosed with pastern dermatitis and eight client-owned unaffected control horses were utilized for this study. METHODS: A cross-sectional study was performed. History and physical examination findings were recorded, and samples were collected and tested for D. congolensis utilizing cytological evaluation and RT-qPCR. Dermatophyte culture and superficial skin scrapings were also performed. RESULTS: Ten of 15 horses with pastern dermatitis had feathered pasterns. Dermatophilus congolensis was identified by RT-qPCR from one nonfeathered horse but none with feathered pasterns. Cytological evaluation identified bacteria in all horses but failed to identify organisms resembling D. congolensis in any horse. Four of 15 horses, all feathered, were positive for Chorioptes mites. Fungal culture was negative for dermatophytes in all horses. All test results were negative for the eight control horses. CONCLUSIONS AND CLINICAL IMPORTANCE: Dermatophilus congolensis was uncommonly associated with pastern dermatitis in horses in this population. However, chorioptic mange was commonly associated with pastern dermatitis in feathered horses and represented an important differential diagnosis for this clinical presentation.

PCR analysis of a prescription vegetarian diet and use in three dogs with cutaneous adverse food reactions.

BACKGROUND: Cutaneous adverse food reaction (CAFR) is diagnosed by performing an elimination diet trial utilizing prescription or home-cooked diets followed by provocative challenge. OBJECTIVES: To report findings of PCR analysis of a prescription vegetarian diet (RCV) for undeclared proteins of animal origin, as well as to describe its utilization for diagnosis and management of dogs suspected of having CAFR. ANIMALS: Three client-owned dogs. METHODS: PCR analysis of RCV for 11 mammalian species and poultry. In three dogs, clinical examination, cytology, aerobic culture (if indicated) and at least one elimination diet trial with RCV. RESULTS: In our case series, all dogs had a history of pruritus and recurrent pyoderma that resolved with infection control and an elimination diet trial. In cases 1 and 2, a diagnosis of CAFR was made following an elimination trial with RCV and provocative challenge. Case 3 had a previously confirmed diagnosis of CAFR and RCV was successfully used to maintain remission of CAFR-related signs. PCR testing of RCV was negative for 11 mammalian species and poultry. CONCLUSIONS AND CLINICAL IMPORTANCE: The RCV diet was found not to contain any undeclared mammalian or avian proteins. In this case series, the RCV was successfully used to diagnose and maintain three dogs with CAFR.

Fluoride release and re-release from a bioactive restorative material.

PURPOSE: To compare the amount of fluoride release and re-release of three different restorative materials. METHODS: The three restorative materials included a resin-based composite (Z100TM, 3M-ESPE), a resin-modified glass ionomer cement (VitremerTM, 3M-ESPE) and a bioactive material (Activa Bioactive-RestorativeTM, Pulpdent,). Ten disks were fabricated from each material. The disks were immersed in deionized water and stored. Samples were taken from each vial on Days 1, 7, 14 and 30 for fluoride ion analysis. Each disk was then exposed to 2.0% neutral sodium fluoride gel (0.9% fluoride ion, Dentsply), immersed in deionized water and stored. Samples were taken on Days 1, 7, 14 and 30 for fluoride ion analysis utilizing a fluoride-specific ion-analyzer. RESULTS: Z100 released less fluoride on Days 1 (P< 0.001), 7 (P= 0.001) and 14 (P< 0.022) for Phase I (initial release) than Phase II (re-release). Vitremer and Activa released less fluoride on Days 7, 14 and 30 (P< 0.001) for Phase II than Phase I. For all intervals of Phase I, Vitremer released the most fluoride, Activa released the second most, and Z100 released the least. These results were the same for Days 7, 14 and 30 of Phase II. The level of fluoride release from Activa was less than that of Vitremer, and greater than that of Z100 for all intervals of Phase I. The results were the same for all but one interval of Phase II. CLINICAL SIGNIFICANCE: This in vitro study evaluated the fluoride release and subsequent re-release of fluoride following a topical fluoride treatment to analyze if the materials were truly bioactive. The results indicate the bioactive material does uptake fluoride and re-release it which could offer inhibition to caries at restoration margins.

Use of mycophenolate mofetil to treat immune-mediated skin disease in 14 dogs - a retrospective evaluation.

BACKGROUND: Mycophenolate mofetil (MMF) is a lymphocytotoxic immunosuppressive agent used in human and companion animal medicine for the treatment of immune-mediated disease. Mycophenolate mofetil is reported to have reduced myelotoxicity and hepatotoxicity when compared to azathioprine. OBJECTIVES: It was hypothesized that treatment with MMF as a secondary agent with glucocorticoids would be effective in treating immune-mediated skin disease. In addition, adverse effects associated with the drug are reported. ANIMALS: Fourteen dogs from a hospital population diagnosed with immune-mediated skin disease. METHODS: A retrospective review of medical records from 2010 to 2015 was used to identify dogs with immune-mediated skin disease that were treated with MMF. RESULTS: All dogs were treated with MMF (mean dose 14.7 mg/kg twice daily) in conjunction with glucocorticoids. Ten of 14 cases showed positive results, with complete remission in eight cases and partial remission in two cases. Mean time to remission was 5.7 weeks. Therapy was discontinued in one case (perianal fistula) due to lack of response. Adverse events were noted in six cases and included diarrhoea (n = 6), haematochezia (n = 2), vomiting (n = 3) and papilloma formation (n = 1). Therapy was discontinued in two cases with diarrhoea. Mycophenolate mofetil was discontinued in an additional case because of a diagnosis of neoplasia. All other adverse events were self-limiting or easily medically managed. No hepatotoxicity or bone marrow suppression was noted. CONCLUSION: This study supports the use of MMF as a second-line immunotherapeutic in immune-mediated skin disease in dogs.

Mycoplasma infection of the middle ear in three cats.

BACKGROUND: Mycoplasma spp. are commensal organisms found in association with the mucus membranes of all mammalian species and are implicated in bacterial infections of many different locations. Mycoplasma spp. as a primary pathogen associated with otitis media in cats has not been reported. OBJECTIVES: To describe three cats with Mycoplasma infection of the middle ear associated with various underlying disease processes. ANIMALS: Three client-owned cats. METHODS: Clinical examination, aerobic culture of the middle ear and computed tomography or magnetic resonance imaging of the skull. RESULTS: Mycoplasma spp. were grown on aerobic culture from the middle ear of three cats. In Case 1, concurrent neoplasia of the bulla was identified. Mycoplasma alone was cultured in Case 2 and Mycoplasma was grown in addition to Bordetella in Case 3. Case 1 was euthanized, Case 2 responded to Mycoplasma targeted therapy and Case 3 responded to Bordetella targeted therapy. CONCLUSIONS AND CLINICAL IMPORTANCE: Mycoplasma infections of the middle ear may be clinically important and require targeted treatment in some cases.

Antibacterial effect of N-acetylcysteine on common canine otitis externa isolates.

BACKGROUND: N-Acetylcysteine (NAC) has the potential to be a useful therapeutic agent for the treatment of otitis externa due to its antimicrobial and mucolytic properties, as well as its ability to disrupt bacterial biofilm. HYPOTHESIS/OBJECTIVES: To determine the antibacterial activity of NAC against common bacterial isolates associated with canine otitis externa. ANIMALS: Twenty two isolates from canine clinical cases of otitis externa were identified and tested, including five Staphylococcus pseudintermedius, six Pseudomonas aeruginosa, five Corynebacterium spp. and six ß-haemolytic Streptococcus spp. isolates. METHODS: Each isolate was grown on blood agar for 24 h and transferred to Mueller Hinton Broth (MHB) to achieve a final concentration of 5 × 10(5)  CFU/mL. NAC was diluted in MHB to a starting concentration of 160 mg/mL and serial two-fold microdilution assays were performed in triplicate with negative controls for all isolates tested. Concentrations of NAC tested ranged from 0.125 to 80 mg/mL. A 50 µL volume of bacterial suspension was used to inoculate each well. RESULTS: The minimum inhibitory concentration (MIC) of NAC for all isolates tested ranged from 5 to 20 mg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: N-Acetylcysteine inhibits clinically relevant and drug resistant bacteria in vitro, and has potential for use as a novel agent for treatment of otitis externa.

Erythema multiforme associated with zonisamide in a dog.

This report describes a dog that developed erythema multiforme in temporal association with administration of the sulphonamide-based anticonvulsant drug zonisamide. Similar adverse drug reactions have been associated with sulphonamide antimicrobial drugs. Caution should be exercised when prescribing this medication for dogs with known hypersensitivity to sulphonamides.

Prevalence of ear disease in dogs undergoing multidetector thin-slice computed tomography of the head.

Previous reports describing the prevalence of ear diseases in dogs have primarily been based on dogs presenting with clinical signs of disease. The prevalence of subclinical ear disease remains unknown. The purpose of this cross-sectional retrospective study was to describe the prevalence of lesions consistent with middle and external ear disease in dogs presented for multidetector computed tomography (CT) of the head and/or cranial cervical spine at our hospital during the period of July 2011 and August 2013. For each included dog, data recorded were signalment, CT findings, diagnosis, and treatment. A total of 199 dogs met inclusion criteria. Nineteen dogs (9.5%) were referred for evaluation of suspected ear disease and 27 dogs (13.5%) had histories or physical examination findings consistent with otitis externa. A total of 163 dogs (81.9%) had CT lesions consistent with external ear disease (i.e. ear canal mineralization, external canal thickening, and/or narrowing of the external canal). Thirty-nine dogs (19.5%) had CT lesions consistent with middle ear disease (i.e. soft tissue attenuating/fluid material in the tympanic bullae, bulla wall thickening or lysis, and/or periosteal proliferation of the temporal bone). Findings from this study indicated that the prevalence of external and middle ear disease in dogs could be higher than that previously reported.

Brief Report: Challenges in Obtaining the Informed Perspectives of Stakeholders Regarding HIV Molecular Epidemiology.

BACKGROUND: HIV molecular epidemiology (HIV-ME) is now being used in a variety of ways, including molecular HIV surveillance to help identify and respond to emerging HIV transmission clusters as specified in the Ending the HIV Epidemic in the U.S. initiative. However, HIV-ME in general, and its use for cluster detection and response, in particular, raises significant ethical and social concerns, which have spurred vigorous debates. Nevertheless, there is a paucity of information regarding how these potential benefits and concerns are perceived among people living with HIV and people without HIV at an increased risk. SETTING: Virtual engagement with US participants. METHODS: We rigorously developed a brief informational video about HIV-ME and conducted a series of in-depth interviews with people living with HIV and people without HIV at an increased risk. RESULTS: Through extensive stakeholder engagement during the video development process and subsequent in-depth interviews (N = 24), several preliminary findings surfaced. In contrast to the high level of concern raised by some critics of HIV-ME, our data appear to show broad support for it. In addition, we observed conflation of perspectives about HIV-ME with concerns about HIV public health surveillance more generally. CONCLUSION: Our experiences reveal substantial communication challenges related to the nature of HIV-ME that need to be overcome to ensure that it is properly understood, which is necessary for meaningfully engaging stakeholders in discussions about its use. Moreover, ongoing, responsive, engagement efforts are critical. Additional systematic data are needed to help inform policy making and practice regarding HIV-ME.

Fast fragment and compound screening pipeline at the Swiss Light Source.

Crystallography-based fragment screening is a highly effective technique employed in structure-based drug discovery to expand the range of lead development opportunities. It allows screening and sorting of weakly binding, low molecular mass fragments, which can be developed into larger high-affinity lead compounds. Technical improvements at synchrotron beamlines, design of innovative libraries mapping chemical space efficiently, effective soaking methods and enhanced data analysis have enabled the implementation of high-throughput fragment screening pipelines at multiple synchrotron facilities. This widened access to CBFS beyond the pharma industry has allowed academic users to rapidly screen large quantities of fragment-soaked protein crystals. The positive outcome of a CBFS campaign is a set of structures that present the three-dimensional arrangement of fragment-protein complexes in detail, thereby providing information on the location and the mode of interaction of bound fragments. Through this review, we provide users with a comprehensive guide that sets clear expectations before embarking on a crystallography-based fragment screening campaign. We present a list of essential pre-requirements that must be assessed, including the suitability of your current crystal system for a fragment screening campaign. Furthermore, we extensively discuss the available methodological options, addressing their limitations and providing strategies to overcome them. Additionally, we provide a brief perspective on how to proceed once hits are obtained. Notably, we emphasize the solutions we have implemented for instrumentation and software development within our Fast Fragment and Compound Screening pipeline. We also highlight third-party software options that can be utilized for rapid refinement and hit assessment.

An ergonomic assessment of the long handle blueberry harvesting rake.

BACKGROUND: Previous work shows the superiority of the long-handled blueberry harvesting rake (LHR) for worker preference and productivity compared to the short-handle rake (SHR). METHODS: Post-shift interviews on occurrence, location, and severity of pain, and video-based observation of body postures enabled ergonomic assessment of Maine workers harvesting blueberries. Workers randomly crossed between LHR and SHR on consecutive work days. Wilcoxon tests compared proportions of specific body postures between LHR and SHR. RESULTS: Subjects used SHR for shorter work periods than LHR. Thirty workers provided interviews for both one LHR and one SHR shift. Assessment of these matched pairs suggested a trend toward less frequent overall pain (P = 0.07) and back pain (P = 0.11) with the LHR versus the SHR. Video tape analysis included 17 sets of observations (8 SHR and 9 LHR) on 12 individuals. Posture assessment showed more severe forward bend and squatting with the SHR and more moderate/neutral postures with the LHR. CONCLUSION: Harvesting with the traditional SHR is likely to be associated with increased frequency of pain in general, and mid-low back pain in particular, when compared to the newer LHR. This may well relate to the work postures associated with each rake. Periodicals, Inc.