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Tinnitus and other audio-vestibular symptoms following COVID-19 vaccination have been described, however, a direct correlation has not been established. The underlying mechanism contributing to this potential vaccine adverse event remains unclear. Our study aims to evaluate the Department of Defense population with tinnitus following immunization.

The accurate control of multicomponent self-assembly to fabricate a single assembly is still a challenge. Herein we describe multicomponent coordination-driven self-assembly with appropriately designed stoichiometric ratios to control the formation of fused C3vpolygons. Each of the assemblies involves two donors and one acceptor molecule, combined in the proper ratios assembling to give two rhomboidal trefoils and two triangular trefoils. The structures were characterized by 1H NMR, 31P NMR, 1H–1H NOESY NMR, DOSY NMR, ESI-MS, DLS, and TEM. Both the rhomboidal trefoils and triangular trefoils exhibit flat conformations on the basis of their calculated structures.

AimsDespite recent advances in arthroscopic rotator cuff repair, re-tear rates remain high. New methods to improve healing rates following rotator cuff repair must be sought. Our primary objective was to determine if adjunctive bone marrow stimulation with channelling five to seven days prior to arthroscopic cuff repair would lead to higher Western Ontario Rotator Cuff (WORC) scores at 24 months postoperatively compared with no channelling.MethodsA prospective, randomized controlled trial was conducted in patients undergoing arthroscopic rotator cuff repair. Patients were randomized to receive either a percutaneous bone channelling of the rotator cuff footprint or a sham procedure under ultrasound guidance five to seven days prior to index surgery. Outcome measures included the WORC, American Shoulder and Elbow Surgeons (ASES), and Constant scores, strength, ultrasound-determined healing rates, and adverse events.ResultsOverall, 94 patients were randomized to either bone channelling or a sham procedure. Statistically significant improvements in all clinical outcome scores occurred in both groups from preoperative to all timepoints (p < 0.001). Intention-to-treat analysis revealed no statistical differences in WORC scores between the two interventions at 24 months postoperatively (p = 0.690). No differences were observed in secondary outcomes at any timepoint and healing rates did not differ between groups (p = 0.186).ConclusionPreoperative bone channelling one week prior to arthroscopic rotator cuff repair was not associated with significant improvements in WORC, ASES, Constant scores, strength, or ultrasound-determined healing rates.Cite this article: Bone Joint J2021;103-B(1):123–130.

Aspergillusspecies cause diverse clinical manifestations in bronchiectasis including allergic bronchopulmonary aspergillosis (ABPA), Aspergillussensitization (AS), and raised IgG indicating exposure to or infection with Aspergillus.

The new bit also saved cost by repeatedly drilling the entire interval in one run. Special Focus: DRILLING Four-blade bit helps reduce drilling time by as much as half The [...]

Immune Thrombocytopenia (ITP) is a rare cause of low platelets in the general population. It is also a rare vaccine side effect. There are few case studies and limited data regarding this relationship.

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For coherent population trapping (CPT) resonance excited by a bichromatic field in a closed Λ-system, we have obtained an analytical expression for the line shape in the case of a Gaussian profile of the radiation intensity distribution, taking into account the light shift of the reference transition frequency. Due to the spatial inhomogeneity of the light shift, the line shape is asymmetric. The dependence of the shift of the resonance peak position on the light intenisty is investigated.

ABSTRACTBovine tuberculosis is a major problem in many countries; hence, new and better diagnostic tools are urgently needed. In this work, we have tested ESAT6, CFP10, PE13, PE5, MPB70, TB10.4, and TB27.4 for their potentials as diagnostic markers in field animals from Northern Ireland, Mexico, and Argentina, regions with low, medium, and high prevalences of bovine tuberculosis, respectively. At all three sites, ESAT6 and CFP10 were superior diagnostic antigens, while their combination performed even better at the two sites where the combination was tested, providing the best coverage for the detection of diseased populations. The high sensitivity in the skin test reactor groups, combined with the high specificity in the tuberculosis-free groups, indicated that a diagnosis could correctly be made for 85% of the infected animals, based on their responses to these two antigens. Furthermore, TB10.4, PE13, and PE5 have the potential to supplement ESAT6 and CFP10 in a future five-component diagnostic cocktail.

ABSTRACTCross-reactive responses elicited by exposure to nontuberculous mycobacteria often confound the interpretation of antemortem tests for Mycobacterium bovisinfection of cattle. The use of specific proteins (e.g., ESAT-6, CFP-10, and MPB83), however, generally enhances the specificity of bovine tuberculosis tests. While genes for these proteins are absent from many nontuberculous mycobacteria, they are present in M. kansasii. Instillation of M. kansasiiinto the tonsillar crypts of calves elicited delayed-type hypersensitivity and in vitro gamma interferon and nitrite concentration responses of leukocytes to M. aviumand M. bovispurified protein derivatives (PPDs). While the responses of M. kansasii-inoculated calves to M. aviumand M. bovisPPDs were approximately equivalent, the responses of M. bovis-inoculated calves to M. bovisPPD exceeded their respective responses to M. aviumPPD. The gamma interferon and nitrite responses of M. kansasii-inoculated calves to recombinant ESAT-6-CFP-10 (rESAT-6-CFP-10) exceeded corresponding responses of noninoculated calves as early as 15 and 30 days after inoculation, respectively, and persisted throughout the study. The gamma interferon and nitrite responses of M. bovis-inoculated calves to rESAT-6-CFP-10 exceeded the corresponding responses of M. kansasii-inoculated calves beginning 30 days after inoculation. By using a lipoarabinomannan-based enzyme-linked immunosorbent assay, specific serum antibodies were detected as early as 50 days after challenge with M. kansasii. By a multiantigen print immunoassay and immunoblotting, serum antibodies to MPB83, but not ESAT-6 or CFP-10, were detected in M. kansasii-inoculated calves; however, responses to MPB83 were notably weaker than those elicited by M. bovisinfection. These findings indicate that M. kansasiiinfection of calves elicits specific responses that may confound the interpretation of bovine tuberculosis tests.

ABSTRACTBovine tuberculosis persists as a costly zoonotic disease in numerous countries despite extensive eradication and control efforts. Sequential serum samples obtained from Mycobacterium bovis-infected cattle were evaluated for seroreactivity to mycobacterial antigens. Animals received M. bovisby aerosol, intratonsil, intranasal, or intratracheal inoculation. Assays included the multiantigen print immunoassay for determination of antigen recognition patterns, immunoblot analysis for sensitive kinetic studies, and the VetTB STAT-PAK test, a novel, rapid test based on lateral-flow technology. Responses to MPB83 were detected for all M. bovis-infected animals regardless of the route or strain of M. bovisused for inoculation. Other less commonly recognized antigens included ESAT-6, CFP-10, and MPB70. Responses to MPB83 were detectable as early as 4 weeks after inoculation, were boosted upon injection of purified protein derivatives for skin testing, and persisted throughout the course of each of the four challenge studies. MPB83-specific immunoglobulin M (IgM) was detected prior to MPB83-specific IgG detection; however, early IgM responses rapidly waned, suggesting a benefit of tests that detect both IgM- and IgG-specific antibodies. The VetTB STAT-PAK test detected responses in sera from 60% (15/25) of the animals by 7 weeks after challenge and detected responses in 96% (24/25) of the animals by 18 weeks. These findings demonstrate the potential for new-generation antibody-based tests for the early detection of M. bovisinfection in cattle.

ABSTRACTDespite having a very low incidence of disease, reindeer (Rangifer tarandus) are subject to tuberculosis (TB) testing requirements for interstate shipment and herd accreditation in the United States. Improved TB tests are desperately needed, as many reindeer are falsely classified as reactors by current testing procedures. Sera collected sequentially from 11 (experimentally) Mycobacterium bovis-infected reindeer and 4 noninfected reindeer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for antibody specific to M. bovisantigens. Specific antibody was detected as early as 4 weeks after challenge with M. bovis. By MAPIA, sera were tested with 12 native and recombinant antigens, which were used to coat nitrocellulose. All M. bovis-infected reindeer developed responses to MPB83 and a fusion protein, Acr1/MPB83, and 9/11 had responses to MPB70. Other antigens less commonly recognized included MPB59, ESAT-6, and CFP10. Administration of purified protein derivatives for skin testing boosted serum antibody responses, as detected by each of the assays. Of the noninfected reindeer, 2/4 had responses that were detectable immediately following skin testing, which correlated with pathological findings (i.e., presence of granulomatous lesions yet the absence of acid-fast bacteria). The levels of specific antibody produced by infected reindeer appeared to be associated with disease progression but not with cell-mediated immunity. These findings indicate that M. bovisinfection of reindeer elicits an antibody response to multiple antigens that can be boosted by skin testing. Serological tests using carefully selected specific antigens have potential for early detection of infections in reindeer.

Objective. To quantify the set-up costs and monetary benefits of a welfare rights service integrated within an NHS service provider, that selects eligible patients using the Health Assessment Questionnaire (HAQ) and offers welfare rights advice to assist in application for Disability Living Allowance and Attendance Allowance.Method. (1) Design: a cost evaluation of a social intervention, screening with the HAQ and welfare rights advice in primary care and hospital settings. (2) Setting: Eight general practices and four hospital rheumatology out-patient departments were selected from four localities in the southwest of England. (3) Participants: Two hundred and sixty-eight eligible patients with arthritis accepted an interview with a welfare rights officer (WRO) from a sample of 1989 service users identified from GPs’ records and hospital out-patient lists. Two hundred and forty two service users expressed an interest in take up of the social intervention. (4) Service users with a HAQ score ≥1.5 were contacted by telephone and offered an appointment with an experienced WRO to help them complete a welfare benefit application form. A ‘micro-costing’ study was undertaken with assessment of monetary benefits received.Results. The indicative set-up costs of similar welfare rights services are £8125 in a GP setting and £9307 per annum in a hospital setting at 2002 prices. Total annual unclaimed Disability Living Allowance/Attendance Allowance granted to successful claimants was £184,382 in the GP setting (n = 84 from 137) and £169,309 in the hospital setting (n = 79 from 131).Conclusions. Welfare rights advice received during a visit to a GP practice or a hospital out-patient department can substantially reduce the level of unclaimed benefit in arthritic populations including the elderly; with mobility and care difficulties. A welfare rights service integrated within a GP practice or hospital that screens people with arthritis using HAQ scores and encourages those with scores ≥1.5 to see a WRO for help with welfare benefit confers monetary benefits for service users that substantially outweigh set-up costs.

ABSTRACTWhite-tailed deer (Odocoileus virginianus) have emerged as reservoirs of bovine tuberculosis in northern America. For tuberculosis surveillance of deer, antibody-based assays are particularly attractive because deer are handled only once and immediate processing of the sample is not required. Sera collected sequentially from 25 Mycobacterium bovis-infected and 7 noninfected deer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for immunoglobulin specific to M. bovisantigens. Various routes of experimental M. bovisinfection, such as intratonsillar inoculation (n= 11), aerosol (n= 6), and exposure to infected deer (in contact, n= 8), were studied. Upon infection, specific bands of reactivity at ~24 to 26 kDa, ~33 kDa, ~42 kDa, and ~75 kDa to M. boviswhole-cell sonicate were detected by immunoblot. Lipoarabinomannan-specific immunoglobulin was detected as early as 36 days postchallenge, and responses were detected for 94% of intratonsillarly and “in-contact”-infected deer. In MAPIA, sera were tested with 12 native and recombinant antigens coated on nitrocellulose. All in-contact-infected (8 of 8) and 10 of 11 intratonsillarly infected deer produced antibody reactive with one or more of the recombinant/native antigens. Responses were boosted by injection of tuberculin for intradermal tuberculin skin testing. Additionally, three of six deer receiving a very low dose of M. bovisvia aerosol exposure produced antibody specific to one or more recombinant proteins. M. boviswas isolated from one of three nonresponding aerosol-challenged deer. Of the 12 antigens tested, the most immunodominant protein was MPB83; however, a highly sensitive serodiagnostic test will likely require use of multiple antigens.

Objectives. To test, in a variety of health settings, the ability of the Health Assessment Questionnaire (HAQ) disability index to predict the eligibility of patients with moderate or severe arthritis for disability living allowance or attendance allowance.Methods. The study included patients from 20 general practices and four hospital out-patient departments across four areas in the southwest of England. Adults with an established diagnosis of rheumatoid arthritis, or osteoarthritis of the hip or knee, and who were not in receipt of Disability Living Allowance (DLA) or Attendance Allowance (AA) were sent an HAQ. Those who scored 1.5 or more were offered an appointment with a welfare advice worker at which they completed an application for DLA or AA. After 3 months they were contacted by the advice worker and asked about the outcome of their applications.Results. Over half of those who completed an HAQ scored 1.5 or over (moderate to severe disability as measured by the HAQ) and were offered advice from experienced welfare benefits advisors. Of these, 87% applied for DLA or AA. Sixty-nine per cent of the applicants were successful. Those scoring 1.75 and over were more likely to be awarded benefit (73% success CLs 67, 79) than people scoring between 1.5 and 1.625 where 55% (CLs 41,69) of applicants were successful.Conclusion. The HAQ was shown to be a good predictor of eligibility for AA or DLA. It can be used, in a variety of health settings, to indicate patients who, with help from an experienced advisor, are likely to gain increased financial help.

ABSTRACTHuman and bovine tuberculosis have long been detected by skin testing with purified protein derivative (PPD), a complex mix of partly denatured mycobacterial antigens with suboptimal specificity. In the present study, skin tests based on ESAT-6, a recombinantly produced antigen highly specific for tuberculosis infection, were investigated. Although ESAT-6 was strongly recognized in vitro and induced high levels of gamma interferon, initial investigations demonstrated that higher doses of ESAT-6 than of PPD were needed to induce substantial delayed-type hypersensitivity reactions. Also, the kinetics of the skin test response differed for the two reagents; PPD showed maximal response at 72 h, but the response to ESAT-6 often peaked later at 96 h. Tests based on an optimized strategy (400 µg of ESAT-6 measured between 72 and 96 h), in cattle infected with Mycobacterium bovis(n= 22) and animals sensitized by exposure to environmental mycobacteria showed ESAT-6 to have a promising diagnostic potential (sensitivity, 82%; specificity, 100%; optimal cutoff, 3 mm), compared with PPD (sensitivity, 86%; specificity, 90%; optimal cutoff, 4 mm). Larger investigations are required to refine cutoff points for any new diagnostic test, but the present results indicate great potential for skin tests based on specific antigens for accurate in vivo diagnosis of tuberculosis.

Background. Patients with rheumatoid arthritis (RA) are traditionally seen regularly as out‐patients, irrespective of whether it is appropriate or timely to see them. A randomized controlled trial has shown that over 2 yr, seeing patients only when they or their general practitioner (GP) request a review saves time and resources and is more convenient. This study aimed to assess clinical and psychological outcomes when the trial was extended to 4 yr.Method. A total of 209 patients were randomized into either ‘routine review’ (control) or ‘no routine follow‐up’ but access to rapid review on request (direct access). Clinical and psychological status and patient satisfaction and confidence were reviewed after 24 and 48 months.Results. Mean age at entry was 56 yr and mean disease duration 11 yr, and 134 patients remained in the study after 48 months. There were no differences between the groups, nor between those who completed the study and those who did not. There were no major differences in clinical or psychological status between the groups at 24 or 48 months. However, self‐efficacy for function was stronger at 48 months for direct access patients (mean 64.0 vs 52.0, P=0.005), as was self‐efficacy for other symptoms (mean 67.8 vs 59.3, P=0.009). Satisfaction at 48 months was increased in direct access compared with control (mean 8.7 vs 7.6, P=0.01) as was confidence in the system (8.9 vs 7.6, P<0.01).Conclusion. It is effective for patients with rheumatoid arthritis to have no regular follow‐up, provided they have access to rapid review when they or their GP request it. Patients using a self‐referral system of care had higher self‐efficacy and greater satisfaction and confidence than those using the traditional system.

The aim of this study was to determine the response of inflammatory and vasoactive mediators to 3 consecutive days of exercise in African-American women with and without sickle cell anemia (SCA). Circulating inflammatory mediators [C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor alpha (TNFα)] were measured before, and vasoactive mediators [endothelin-1 (ET-1), nitric oxide metabolites (NOx)] before and after each exercise bout in ten subjects with SCA and ten controls. Exercise did not affect ET-1, IL-6 or CRP concentrations (p>.05). TNFα was higher in SCA than controls (p<.0005) at all times; however, the response pattern was similar for the groups: no change from day I to day 2, but a decrease from day 2 to day 3 (p≤.05). NOx increased significantly after exercise (p≤.0001) but returned to baseline by 24 h afterward. On the 3rd day, NOx increased after exercise in SCA but not in the controls (p≤.05). In conclusion, exercise did not cause a harmful inflammatory response in these individuals with SCA. However, NOx increased after exercise on all 3 days in SCA but appeared attenuated after 2 days in controls.

Exercise training produces enhanced nitric oxide (NO)-dependent, endothelium-mediated vasodilator responses of porcine coronary arterioles but not conduit coronary arteries. The purpose of this study was to test the hypothesis that exercise training increases the amount of endothelial NO synthase (eNOS) in the coronary arterial microcirculation but not in the conduit coronary arteries. Miniature swine were either exercise trained or remained sedentary for 16–20 wk. Exercise-trained pigs exhibited increased skeletal muscle oxidative capacity, exercise tolerance, and heart weight-to-body weight ratios. Content of eNOS protein was determined with immunoblot analysis in conduit coronary arteries (2- to 3-mm ID), small arteries (301- to 1,000-μm ID), resistance arteries (151- to 300-μm ID), and three sizes of coronary arterioles [large (101- to 150-μm ID), intermediate (51- to 100-μm ID), and small (<50-μm ID)]. Immunoblots revealed increased eNOS protein in some sizes of coronary arteries and arterioles but not in others. Content of eNOS was increased by 60–80% in small and large arterioles, resistance arteries, and small arteries; was increased by 10–20% in intermediate-sized arterioles; and was not changed or decreased in conduit arteries. Immunohistochemistry revealed that eNOS was located in the endothelial cells in all sizes of coronary artery. We conclude that exercise training increases eNOS protein expression in a nonuniform manner throughout the coronary arterial tree. Regional differences in shear stress and intraluminal pressures during exercise training bouts may be responsible for the distribution of increased eNOS protein content in the coronary arterial tree.

The identity of the lymphocyte subtypes constituting the lymphocytic mantle within developing early-stage lesions of bovine tuberculosis was investigated immunohistochemically in calves inoculated intranasally with 2×10⁷colony-forming units of a field isolate of Mycobacterium bovis. Pulmonary lesions were examined 7, 14, 21, 28 and 42 days after inoculation, and bronchial lymph node lesions at 35 days. The immunolabelling results reported were obtained with monoclonal antibodies against two T-cell epitopes (WC1⁺γδ and CD2⁺) and against B-cell epitopes. Large numbers of CD2⁺T-lymphocytes were observed around developing areas of necrosis throughout the study; WC1⁺γδ cells, however, were more numerous at these sites up to and including day 21. On the other hand, aggregates of B lymphocytes did not become prominent in areas adjacent to lesions until day 42. The results suggest that these lymphocyte phenotypes play a role in the pathogenesis of early-stage lesions. Copyright Harcourt Publishers Ltd

Human male volunteers were studied in a metabolic facility whilst they were fed randomized controlled diets. In eight volunteers there was a significant increase in faecal apparent total N:-nitroso compounds (ATNC) and nitrite excretion (P < 0.0001 and P = 0.046, respectively) when randomized doses of meat were increased from 0 to 60, 240 and 420 g/day over 10 day periods. Mean (+/- SE) faecal ATNC levels were 54 +/- 7 microg/day when the diets contained no meat, 52 +/- 11 microg/day when the diets contained 60 g meat/day, 159 +/- 33 microg/day with 240 g meat and 199 +/- 36 microg/day with 420 g meat. Higher concentrations of NOC were associated with longer times of transit in the gut (r = 0.55, P = 0.001) and low faecal weight (r = -0.51, P = 0.004). There was no significant decline in levels in individuals fed 420 g meat for 40 days. The exposures found on the higher meat diets were comparable with other sources of N:-nitroso compounds (NOC), such as tobacco smoke. Many NOC are known large bowel initiators and promotors in colon cancer, inducing G-->A transitions in codons 12 and 13 of K-ras. Endogenous NOC formation, combined with prolonged transit times in the gut, may explain the epidemiological associations between high meat/low fibre diets and colorectal cancer risk.

The continued use of purified protein derivative (PPD) tuberculin is considered to be the main factor which limits the specificity of diagnostic tests for bovine tuberculosis (TB). This study evaluated a whole blood interferon‐gamma (IFN‐y) assay and compared the diagnostic potential of PPD with two tuberculosis‐specific antigens, ESAT‐6 and MPB70. To provide estimates of sensitivity and specificity, responses were measured in 180 skin test‐reacting cattle, of which 131 were confirmed as tuberculous, and in 128 cattle from TB‐free herds. For the skin test reactors, there was a positive correlation between the IFN‐y responses to PPD from Mycobacterium bovis (PPDB) and PPD from Mycobaderium avium (PPDA), indicating cross‐reactivity between these conmplex antigens which are the basis of the skin test. In comparisons of the ESAT‐6 IFN‐y test with a PPD IFN‐y test (using PPDB compared with PPDA), there was a decrease in sensitivity (76‐3 per cent vs 89‐3 per cent), but a clear increase in specificity (99‐2 per cent vs 92‐2 per cent).

Tests based on tuberculin purified protein derivative (PPD) cannot distinguish between tuberculosis infection, Mycobacterium bovis BCG vaccination, or exposure to environmental mycobacteria. The present study investigated the diagnostic potential of two Mycobacterium tuberculosis-specific antigens (ESAT-6 and CFP10) in experimental animals as well as during natural infection in humans and cattle. Both antigens were frequently recognized in vivo and in vitro based on the induction of delayed-type hypersensitivity responses and the ability to induce gamma interferon production by lymphocytes, respectively. The combination of ESAT-6 and CFP10 was found to be highly sensitive and specific for both in vivo and in vitro diagnosis. In humans, the combination had a high sensitivity (73%) and a much higher specificity (93%) than PPD (7%).

Protective immunity against tuberculosis is considered to be essentially cell mediated, and an important role for CD8(+) T lymphocytes has been suggested by several studies of murine and human infections. The present work, using an experimental model of infection with Mycobacterium bovis in cattle, showed that live M. bovis elicits the activation of CD8(+) T cells in vitro. However, a sonic extract prepared from M. bovis (MBSE) and protein purified derivative (PPDb) also induced a considerable degree of activation of the CD8(+) T cells. Analysis of proliferative responses of peripheral blood mononuclear cells, purified CD8(+) T cells, and CD8(+) T-cell clones to M. bovis and to soluble antigenic preparations (MBSE, PPDb) showed that the responses of all three types of cells were always superior for live mycobacteria but that strong responses were also obtained with complex soluble preparations. Furthermore, while cytotoxic capabilities were not investigated, the CD8(+) T cells were found to produce and release gamma interferon in response to antigen (live and soluble), which indicated one possible protective mechanism for these cells in bovine tuberculosis. Finally, it was demonstrated by metabolic inhibition with brefeldin A and cytochalasin D at the clonal level that an endogenous pathway of antigen processing is required for presentation to bovine CD8(+) cells and that presentation is also dependent on phagocytosis of the antigen.

Since doses to skin of patients from fluoroscopically-guided interventional procedures can be very high, real-time monitoring of skin dose is important for both patient management and quality control. The use of a scintillation detector, placed on the X-ray port to measure potential skin dose, was investigated, focusing on the uncertainties related to the technique. Sources of uncertainty include performance characteristics of the dosemeter, errors in calibration, patient set-up and changes during the procedure. Some of the largest sources of error include uncertainty in source-to-skin distance, heel effect, difficulty in identifying the area of skin principally exposed, calibration error, energy dependence of the dosemeter and the dose rate dependence of the monitor. This technique is found to be beneficial for radiation management, but users must be cognizant of the potential errors of the method and the limitations that these place on quality control and patient management. Knowing the limitations and minimizing the sources of error enhance the utility of the technique.

Tuberculosis continues to be a worldwide problem for both humans and animals. The development of tests to differentiate between infection with Mycobacterium tuberculosis or Mycobacterium bovis and vaccination with M. bovis BCG could greatly assist in the diagnosis of early infection as well as enhance the use of tuberculosis vaccines on a wider scale. Recombinant forms of four major secreted proteins of M. bovis-MPB59, MPB64, MPB70, and ESAT-6-were tested in a whole-blood gamma interferon (IFN-gamma) assay for differentiation between cattle vaccinated with BCG and those experimentally infected with M. bovis. BCG vaccination induced minimal protection in the present study, with similar numbers of animals infected with M. bovis in BCG-vaccinated and nonvaccinated groups. Following vaccination with BCG, the animals produced moderate IFN-gamma responses to bovine purified protein derivative (PPDB) but very weak responses to the recombinant antigens. Cattle from both the BCG-vaccinated and nonvaccinated groups which were M. bovis culture positive following challenge produced IFN-gamma responses to PPDB and ESAT-6 which were significantly stronger than those observed in the corresponding M. bovis culture-negative animals. IFN-gamma responses to MPB59, MPB64, and MPB70 were significantly weaker, and these antigens could not discriminate between vaccinated animals which develop disease and the culture-negative animals. The results of the study indicate that of the four antigens tested in the IFN-gamma assay, only ESAT-6 would be suitable for differentiating BCG-vaccinated animals from those infected with bovine tuberculosis.

The whole cell patch clamp technique has been used to record Ca2+-activated cation and chloride conductances evoked by release of Ca2+from intracellular stores of cultured neonatal dorsal root ganglion neurones. The aim of this study was to investigate metabotropic glutamate receptor (mGluR) mechanisms and evaluate a possible role for cyclic ADP-ribose as an intracellular signalling molecule. Glutamate and the metabotropic glutamate receptor agonist (1S,3R)-ACPD-evoked transient depolarizations, Ca2+-activated inward currents and rises in intracellular Ca2+. The (1S,3R)-ACPD-activated currents were insensitive to InsP3signalling inhibitors, heparin and pentosan polysulphate. Intracellular application of ryanodine alone activated currents in this study and proved a difficult tool to use as a potential inhibitor of cyclic ADP-ribose-mediated responses. However, intracellular dantrolene did attenuate both (1S,3R)-ACPD and cyclic ADP-ribose responses. Intracellular photorelease of cGMP and cyclic ADP-ribose mimicked the responses to mGluR receptor activation. Intracellular application of nicotinamide and W7 inhibited the responses to photoreleased cGMP but did not prevent responses to mGluR activation. The cyclic ADP-ribose receptor antagonist 8-amino cyclic ADP-ribose attenuated responses to (1S,3R)-ACPD, cGMP and cyclic ADP-ribose, but some Ca2+-activated inward currents were still observed in the presence of this antagonist. In conclusion, mGluR receptor activation, cGMP and cyclic ADP-ribose release Ca2+from intracellular stores. Some evidence suggests that pharmacologically related pathways are involved.

The hazards of dealing with infected false femoral aneurysms resulting from intra-arterial narcotic injection are highlighted in six patients. Two patients were human immunodeficiency virus positive and three patients were hepatitis B surface antigen positive. Because of these infections exploration of groin swellings as presumed soft tissue abscesses is potentially hazardous without proper proximal vascular control. All patients underwent reconstruction following arterial ligation and five grafts became infected, with life threatening haemorrhage occurring in four patients. Five grafts have subsequently occluded or have been removed without loss of limb viability, although two patients have been regrafted. A high index of suspicion and assessment by a vascular surgeon, with angiography if indicated, is required in any intravenous drug abuser presenting with a groin swelling following injection. Because of the great risk of graft infection, it is suggested that ligation and debridement alone be carried out, with immediate arterial reconstruction only for non-viability.

Seventy consecutive patients with infrainguinal bypass grafts entered a 1-year graft surveillance programme involving colour duplex scanning, direct graft insonation and computer-assisted impedance analysis. Graft patients with a positive duplex scan, high frequencies on graft insonation or an impedance value above 0.50 subsequently underwent arteriography. Sixteen patients were excluded before the initial surveillance visit. The 54 remaining patients with grafts (30 vein, 24 synthetic) underwent a total of 137 surveillance visits, with 21 grafts confirmed to be ‘at risk’. The sensitivity of an impedance value above 0.55 in identifying these grafts was 86 per cent, rising to 95 per cent when combined with graft insonation. Duplex scanning did not identify any abnormalities in 11 grafts that were either shown by arteriography to be ‘at risk’ or occluded before arteriography. Impedance measurement and graft insonation are simple screening techniques with a high sensitivity (when combined), which identify ‘at risk’ infrainguinal grafts. Positive graft insonation or an impedance value over 0.55 will identify all ‘at risk’ vein grafts while minimizing the number of unnecessary arteriograms.

Canine abdominal aortas have been replaced with Dacron® arterial prostheses to assess the effects of mesothelial cell seeding on graft prostacyclin and thromboxane A2release. At both 2 weeks and 6 weeks after surgery, three seeded and two unseeded control grafts were examined for prostacyclin release. In addition, thromboxane release was assessed in one seeded and one unseeded graft. Sections of aorta and graft were removed and incubated in PBS containing either 10 μM calcium ionophore A23187 or 20 μM arachidonic acid. The incubation mixture was sub-sampled at 5 min intervals over a 20 min period to assess the progressive release of prostacyclin and thromboxane A2using a radioimmunoassay for 6-keto-prostaglandin F1αand thromboxane B2respectively. In seeded grafts, 6-keto-prostaglandin F1αrelease averaged 15 per cent compared with aorta at 2 weeks and 45 per cent compared with aorta at 6 weeks. By contrast, release from unseeded grafts was undetectable at 2 weeks; however, by 6 weeks there was some release amounting to 15 per cent compared with aorta. There was a statistically significant increase in the release of 6-keto-prostaglandin F1αfrom mesothelial cell seeded grafts at 6 weeks compared with unseeded grafts (P< 0·01). Thromboxane release from the graft sections was variable and unrelated to whether the grafts had been seeded or not. These preliminary results, showing that grafts seeded with autologous peritoneal mesothelial cells release more prostacyclin than unseeded grafts, further highlight the role of the mesothelial cell as an alternative to the endothelial cell for improving the patency of arterial Dacron prostheses in the early postoperative days.

Fifty minor foot and transmetatarsal amputations were studied to assess the reliability of Doppler ankle blood pressure (DABP) and skin blood flow (SBF) to predict healing. The level of amputation was determined solely on clinical criteria. Thirty-six (72 per cent) of the amputations healed. There was no statistical difference between mean DABP in healed 89±8 mmHg mean ± s.e.m.) and non-healed (91 ± 12 mmHg mean ± s.e.m.) amputations. SBF was assessed by 125I-iodoantipyrine clearance in 28 patients. There was no correlation between DABP and SBF (r = 0.038). SBF in patients with healed amputations was 14.8 ± 1.2 (mean ± s.e.m.) ml 100 g−1min−1in contrast to SBF of 5.9 ± 0.3 (mean ± s.e.m.) ml 100 g−1min−1in the patients with non-healing (P<0.01). Where the flow was in excess of 8 ml 100 g−1min−1healing was always obtained while a flow of less than 7 ml 100 g−1min−1was associated with healing failure. These results suggest that DABP should be interpreted with caution as this technique fails as an accurate means of identifying those patients suitable for forefoot amputations. Skin blood flow assessment appears to provide an absolute value for the prediction of healing potential at this level.

Non-atheromatous stenosis of the subclavian artery is rare. A 10-year experience, comprising 5 cases of non-infective arteritis and 1 of fibromuscular dysplasia, in patients of average age 29 years, is described. The literature is reviewed and the possible aetiology discussed. Acceptable functional results were obtained using extra-anatomical procedures such as carotid subclavian and carotid axillary bypass. There was no appreciable morbidity.

A hypermutable strain of Drosophila simulans that originated from a single spontaneous mutant male was characterized. Seven different mutations were isolated from roughly 100 generations of offspring. The genetic analysis of the viable mutants showed two mutations on the X chromosome, one in the lozenge locus and the other in the ruby gene. The autosomic mutations characterized were a dpp-held out-like, a blistered-like and a homoeotic dominant mutant with an antenna-to-leg transformation and ectopic eyes that we called Zoinho-napata.

Using two-dimensional gel electrophoresis, we have analyzed proteins from a plasma membrane-enriched fraction from Phycomyces sporangiophores. Specifically, we have compared gels for night-blind mutants and a wild-type strain to find proteins involved in the early steps of the sensory transduction chain for phototropism. In the gels for a mutant affected in the gene madA, a protein spot [51 kilodaltons (kdal) and pI 6.35] appears that is absent from the wild-type and the other mad mutants. Mutants affected in either of two madB alleles lack a protein spot (57 kdal and pI 6.6) that is present in the wild-type and all other mad strains; this spot probably represents the madB gene product. In some madC mutants, two spots (59 kdal, pI 6.5, with a covalently linked flavin; and 50 kdal, pI 6.4) are absent; however, in other madC strains, one or both of these spots are present. These four protein spots that are altered in madA, madB, and madC mutants may represent components of the photoreceptor complex responsible for phototropism in Phycomyces.

Synthetic homologous histidine peptides were found to directly and irreversibly inactivate herpes simplex virus types 1 and 2 (HSV-1 and HSV-2). The inactivation, which occurred within 1 min of virus exposure to the drug, was independent of temperature but dependent upon the pH and molecular size of the polypeptide. Poly-L-histidine consisting of 24 residues (His-24), with a molecular weight (m.w.) of 3,310, inactivated greater than 99% of the virus present at pH 6.0 and 62% at pH 5.0. Poly-L-histidine consisting of 64 (His-64; average m.w., 8,800) or 75 residues (His-75; average m.w., 10,300) inactivated greater than 99% of virus present at pH 5.0 and 6.0. However, His-24, -64, and -75 were not active against these viruses at pH 7.0 or 8.0. At the concentrations tested, poly-L-histidine of 12 (His-12; m.w., 1,665) or 18 (His-18; m.w., 2,487) residues had no effect on HSV-1 or HSV-2 at any of the pHs tested. When these studies were repeated with other basic homologous polypeptides, poly-L-arginine and poly-L-lysine, various degrees of inactivation were observed that were most pronounced in the neutral-to-alkaline pH range. Once virus was inactivated by poly-L-histidine, the effect could not be reversed in vitro by raising the pH to 7.2 or in vivo by injecting the virus into the neutral environment of an animal. These data suggest that the presence of histidine residues in a peptide of suitable structure may endow that peptide with potent antiviral capabilities.

Chemical modification of the gamma-carboxyglutamyl (Gla) residues of bovine prothrombin fragment 1 using the formaldehyde-morpholine method in the presence of 100 Kappm Tb3+ ions at pH 5.0 provided a modified protein containing 3 gamma-methyleneglutamyl residues (gamma-MGlu) and 7 Gla residues (bovine 3-gamma-MGlu-fragment 1). The modified protein bound the same number of Ca2+ ions as the native protein (six to seven), exhibited 28Mg2+-binding properties identical to native fragment 1 (five Mg2+ ions bound), exhibited the metal ion-promoted quenching of the intrinsic fluorescence in a manner similar to the native protein, but did not bind to phosphatidylserine (PS)/phosphatidylcholine (PC) vesicles in the presence of Ca2+ ions. Modification of the bovine protein using [14C]formaldehyde-morpholine provided a 14C-labeled 3-gamma-MGlu-fragment 1 suitable for sequence analysis. Edman sequencing of the peptides released by a tryptic digest of the reduced and carboxymethylated bovine [14C]3-gamma-MGlu-fragment 1 indicated that Gla residues at positions 7, 8, and 33 had been converted to [14C]gamma-methyleneglutamyl residues. In addition Lys97 was found to contain a 14C label. Similar analysis of the human [14C]3-gamma-MGlu-fragment 1 indicated that Gla residues at positions 7 and 32 were major modification sites and that Gla residues at positions 6 and 14 were partially modified. Lysine 96 was also modified in the human protein. The incorporation of a 14C label at Lys97 in bovine 3-gamma-MGlu-fragment 1 protein is not responsible for the loss of Ca2+-promoted binding to PS/PC vesicles. We suggest that Gla residues 7, 8, and 33 are elements of the first Ca2+-binding site; occupancy of this site establishes the Ca2+-specific conformation which is essential for the Ca2+-promoted interaction of the bovine protein with PS/PC vesicles. These studies also suggest that the loss of Gla residues at positions 7 and 32 prevents the formation of the initial Ca2+-binding site in the human protein.