Quantitative and equipment-free paper-based agglutination assay of bacterial cells.

Background: point-of-care (POC) tests are useful for bedside/home applications, emergencies, frequent follow-ups, and resource-limited areas. Limited quantitative and equipment-free POC assays have been reported. This study aims to develop, validate, and apply a simple, quantitative, paper-based POC assay. Methods: wax-channeled paper treated with specific anti-Brucella and anti-Salmonella antibodies was used for distance-based chromatographic elution of stained bacterial cell agglutinations. Results: a qualitative paper-based agglutination POC test was developed using color intensity, tail appearance, and "+/-" signs that clearly distinguish the positive and negative results. The optimization of the test for paper type, microfluidic channel design, antibody and bacterial cell concentrations, and elution methods was carried out. Quantitative assay transformation was successfully developed using the color intensity of the original reaction zone, intensity of elution tail, and distance-based migration that correspond to bacterial agglutination size. The migration distance of eluted bacterial agglutination bands corresponds to the target concentration with good linearity and minimal variability. Reporting of colored band migration with numbers using microfluidic patterns was used to enhance non-technical end-user applications. A distance-based POC assay prototype was then successfully used for the accurate detection of known and unknown samples in comparison with standard assays. Conclusions: the migration distance of an eluted stained bacterial agglutination correlated with anti-bacterial antibody concentrations. A simple, cheap, quantitative, and equipment-free paper-based POC assay of bacterial cell agglutination was developed. This test can be used for simple "+/-" results, thermometer-like quantification, or text reporting with numbers corresponding to target concentrations. The assay has extended applications to different human disease biomarkers.

The Influence of Unstable Load and Traditional Free-Weight Back Squat Exercise on Subsequent Countermovement Jump Performance.

The purpose of the present study was to examine the effects of a back squat exercise with unstable load (UN) and traditional free-weight resistance (FWR) on subsequent countermovement jump (CMJ) performance. After familiarisation, thirteen physically active males with experience in resistance training visited the laboratory on two occasions during either experimental (UN) or control (FWR) conditions separated by at least 72 h. In both sessions, participants completed a task-specific warm-up routine followed by three maximum CMJs (pre-intervention; baseline) and a set of three repetitions of either UN or FWR back squat exercise at 85% 1-RM. During the UN condition, the unstable load was suspended from the bar with elastic bands and accounted for 15% of the total load. Post-intervention, three maximum CMJs were performed at 30 s, 4 min, 8 min and 12 min after the last repetition of the intervention. The highest CMJ for each participant was identified for each timepoint. No significant increases (p > 0.05) in jump height, peak concentric power, or peak rate of force development (RFD) were found after the FWR or UN conditions at any timepoint. The lack of improvements following both FWR and UN conditions may be a consequence of the low percentage of unstable load and the inclusion of a comprehensive task-specific warm-up. Further research is required to explore higher UN load percentages (>15%) and the chronic effects following the implementation of a resistance training programme.

Supporting Smokers in Difficult Settings: Suggestions for Better Education and Counseling in Cancer Centers in Jordan.

Continued smoking in cancer patients is commonly observed in Jordan. In a country that exhibits some of the highest smoking rates globally, enhancing patient education regarding the value of smoking cessation for cancer care is vital. The objectives of our study were to describe sociodemographic and clinical factors associated with continued smoking in Jordanian smokers after a cancer diagnosis; to identify reasons for smoking and knowledge regarding smoking's impact on care; to examine in a multivariable manner the factors associated with continued smoking, and to accordingly generate patient counseling recommendations. An interviewer-administered survey using the Theoretical Domains Framework was employed. Among 350 subjects (mean age 51.0, median 52.7), approximately 38% of patients had quit or were in the process of quitting; 61.7% remained smokers. Substantial knowledge gaps with regard to the impact of continued smoking on cancer care were observed. Remaining a smoker after diagnosis was associated with being employed, not receiving chemotherapy or surgery, having lower confidence in quitting, and having a lower number of identified reasons for smoking. Interventions to promote cessation in Jordanian cancer patients who smoke should focus on enhancing patient awareness about the impact of smoking in cancer care and raising perceived self-efficacy to quit.

Feasibility, Psychosocial Effects, Influence, and Perception of Elastic Band Resistance Balance Training in Older Adults.

This study utilised feedback from older adults during balance-challenging, elastic band resistance exercises to design a physical activity (PA) intervention. METHODS: Twenty-three active participants, aged 51-81 years, volunteered to perform a mini balance evaluation test and falls efficacy scale, and completed a daily living questionnaire. Following a 10 min warm-up, participants performed eight pre-selected exercises (1 × set, 8-12 repetitions) using elastic bands placed over the hip or chest regions in a randomised, counterbalanced order with 15 min seated rests between interventions. Heart rate (HR) and rate of perceived exertion (RPE) were measured throughout. Participant interview responses were used to qualify the experiences and opinions of the interventions including likes, dislikes, comfort, and exercise difficulty. RESULTS: Similar significant (p < 0.01) increases in HR (pre- = 83-85 bpm, mid- = 85-88 bpm, post-intervention = 88-89 bpm; 5-6%) and RPE (pre- = 8-9, mid- = 10, post-intervention = 10-11) were detected during the PA interventions (hip and chest regions). Interview data revealed that participants thought the PA interventions challenged balance, that the exercises would be beneficial for balance, and that the exercises were suitable for themselves and others. Participants reported a positive experience when using the PA interventions with an elastic band placed at the hip or chest and would perform the exercises again, preferably in a group, and that individual preference and comfort would determine the placement of the elastic band at either the hip or chest. CONCLUSION: These positive outcomes confirm the feasibility of a resistance band balance program and will inform intervention design and delivery in future studies.

Cessation experiences and quitting perspectives of Jordanian cancer patients who smoke.

OBJECTIVES: To describe quitting experiences of cancer patients in a Cancer Center in Jordan; to study patients' perceptions regarding the process of smoking cessation; and to provide insights about patients in this difficult setting in order to inform oncology practitioners with regards to how improve perceptions and skills related to quitting. METHODS: An Arabic cross-sectional questionnaire was developed to evaluate smoking and quitting behaviors in the context of cancer. The tool used as its framework the Theoretical Domains Framework to capture quitting perceptions of cancer patients who smoke, as well as social, environmental, and system-level factors that influence quitting. Eligible patients who were treated at the Center (both in-patient and out-patient settings) and who were current smokers or who smoked up to the time of cancer diagnosis were eligible. Patients were interviewed between July, 2018 and January 2020 using two versions of the questionnaire: an 'ex-smokers' version, and a 'current smokers' version. RESULTS: Only a third of subjects (104/350) had been smoke-free for at least 30 days. Both smokers and ex-smokers generally felt that quitting was important, but mean importance and confidence scores (out of 10) were significantly lower in current smokers (8.2 versus 9.1, p-value=0.002; 6.4 versus 8.7, p-value=0.000). Roughly 31% of subjects believed smoking harms were exaggerated and that smoking was not an addiction. About 62% of subjects agreed quitting required skills, and 78.5% felt the steps to quit were clear, but across several listed strategies for quitting, use of these was limited (even in ex-smokers). Among current smokers, roughly a third exhibited forms of cessation fatigue. CONCLUSION: Jordanian cancer patients who smoke present with limited knowledge about the quitting process. Even when some success is observed, low rates of utilization of specific quitting strategies were observed, highlighting the need for better counseling about quitting.

Altered Drop Jump Landing Biomechanics Following Eccentric Exercise-Induced Muscle Damage.

Limited research exists in the literature regarding the biomechanics of the jump-landing sequence in individuals that experience symptoms of muscle damage. The present study investigated the effects of knee localized muscle damage on sagittal plane landing biomechanics during drop vertical jump (DVJ). Thirteen regional level athletes performed five sets of 15 maximal eccentric voluntary contractions of the knee extensors of both legs at 60°/s. Pelvic and lower body kinematics and kinetics were measured pre- and 48 h post-eccentric exercise. The examination of muscle damage indicators included isometric torque, muscle soreness, and serum creatine kinase (CK) activity. The results revealed that all indicators changed significantly following eccentric exercise (p < 0.05). Peak knee and hip joint flexion as well as peak anterior pelvic tilt significantly increased, whereas vertical ground reaction force (GRF), internal knee extension moment, and knee joint stiffness significantly decreased during landing (p < 0.05). Therefore, the participants displayed a softer landing pattern following knee-localized eccentric exercise while being in a muscle-damaged state. This observation provides new insights on how the DVJ landing kinematics and kinetics alter to compensate the impaired function of the knee extensors following exercise-induced muscle damage (EIMD) and residual muscle soreness 48 h post-exercise.

Variable, but not free-weight, resistance back squat exercise potentiates jump performance following a comprehensive task-specific warm-up.

Studies examining acute, high-speed movement performance enhancement following intense muscular contractions (frequently called "post-activation potentiation"; PAP) often impose a limited warm-up, compromizing external validity. In the present study, the effects on countermovement vertical jump (CMJ) performance of back squat exercises performed with or without elastic bands during warm-up were compared. After familiarization, fifteen active men visited the laboratory on two occasions under randomized, counterbalanced experimental squat warm-up conditions: (a) free-weight resistance (FWR) and (b) variable resistance (VR). After completing a comprehensive task-specific warm-up, three maximal CMJs were performed followed by three back squat repetitions completed at 85% of 1-RM using either FWR or VR Three CMJs were then performed 30 seconds, 4 minutes, 8 minutes, and 12 minutes later. During CMJ trials, hip, knee, and ankle joint kinematics, ground reaction force data and vastus medialis, vastus lateralis, and gluteus maximus electromyograms (EMG) were recorded simultaneously using 3D motion analysis, force platform, and EMG techniques, respectively. No change in any variable occurred after FWR (P > 0.05). Significant increases (P < 0.05) were detected at all time points following VR in CMJ height (5.3%-6.5%), peak power (4.4%-5.9%), rate of force development (12.9%-19.1%), peak concentric knee angular velocity (3.1%-4.1%), and mean concentric vastus lateralis EMG activity (27.5%-33.4%). The lack of effect of the free-weight conditioning contractions suggests that the comprehensive task-specific warm-up routine mitigated any further performance augmentation. However, the improved CMJ performance following the use of elastic bands is indicative that specific alterations in force-time properties of warm-up exercises may further improve performance.

Oxytocin and Pain Perception: From Animal Models to Human Research.

An accumulating body of evidence suggests that the hypothalamic neuropeptide oxytocin (OT) has a modulatory effect on pain processing. Particularly strong evidence comes from animal models. Here, we review recent advances in animal research on the analgesic effects of OT and discuss possible target sites of OT within descending and ascending pain pathways in the brain. In addition to the spinal cord being a direct target of the neuropeptide, OT has also been shown to modulate the neuronal activity of limbic and cortical brain regions, which play a major role in the cognitive and emotional processing of pain. Human studies investigating the influence of OT on pain perception are less numerous and have revealed less consistent results. The human literature is therefore scanned thoroughly and different approaches to study the effects of OT on pain perception in humans are discussed. Moreover, we also address how OT might alleviate pain by influencing socio-emotional components in humans. We conclude that further investigating specific OT and OT-sensitive circuits, which modulate pain processing especially in primates, will improve our understanding of OT-analgesic effects. In human research, the increased use of neuroimaging and autonomic measures might help to bridge the gap to animal studies.

A sled push stimulus potentiates subsequent 20-m sprint performance.

OBJECTIVES: The objective of this study was to examine the potentiating effects of performing a single sprint-style sled push on subsequent unresisted 20m sprint performance. DESIGN: Randomized crossover design. METHODS: Following a familiarization session, twenty rugby league players performed maximal unresisted 20m sprints before and 15s, 4, 8 and 12min after a single sled push stimulus loaded with either 75 or 125% body mass. The two sled push conditions were performed in a randomized order over a one-week period. The fastest sprint time recorded before each sled push was compared to that recorded at each time point after to determine the post-activation potentiation (PAP) effect. RESULTS: After the 75% body mass sled push, sprint time was 0.26±1.03% slower at the 15s time point (effect size [ES]=0.07) but faster at the 4 (-0.95±2.00%; ES=-0.22), 8 (-1.80±1.43%; ES=-0.42) and 12 (-1.54±1.54%; ES=-0.36)min time points. Sprint time was slower at all the time points after the 125% body mass sled (1.36±2.36%-2.59±2.90%; ESs=0.34-0.64). CONCLUSIONS: Twenty-meter sprint performance is potentiated 4-12min following a sled push loaded with 75% body mass while it is impaired after a 125% body mass sled. These results are of great importance for coaches seeking to potentiate sprint performance with the sled push exercise.

Chain-loaded variable resistance warm-up improves free-weight maximal back squat performance.

The acute influence of chain-loaded variable resistance exercise on subsequent free-weight one-repetition maximum (1-RM) back squat performance was examined in 16 recreationally active men. The participants performed either a free-weight resistance (FWR) or chain-loaded resistance (CLR) back squat warm-up at 85% 1-RM on two separate occasions. After a 5-min rest, the participants attempted a free-weight 1-RM back squat; if successful, subsequent 5% load additions were made until participants failed to complete the lift. During the 1-RM trials, 3D knee joint kinematics and knee extensor and flexor electromyograms (EMG) were recorded simultaneously. Significantly greater 1-RM (6.2 ± 5.0%; p < .01) and mean eccentric knee extensor EMG (32.2 ± 6.7%; p < .01) were found after the CLR warm-up compared to the FWR condition. However, no difference (p > .05) was found in concentric EMG, eccentric or concentric knee angular velocity, or peak knee flexion angle. Performing a CLR warm-up enhanced subsequent free-weight 1-RM performance without changes in knee flexion angle or eccentric and concentric knee angular velocities; thus a real 1-RM increase was achieved as the mechanics of the lift were not altered. These results are indicative of a potentiating effect of CLR in a warm-up, which may benefit athletes in tasks where high-level strength is required.

Postactivation Potentiation of Horizontal Jump Performance Across Multiple Sets of a Contrast Protocol.

Seitz, LB, Mina, MA, and Haff, GG. Postactivation potentiation of horizontal jump performance across multiple sets of a contrast protocol. J Strength Cond Res 30(10): 2733-2740, 2016-This study determined whether a postactivation potentiation (PAP) effect could be elicited across multiple sets of a contrast PAP protocol. Fourteen rugby league players performed a contrast PAP protocol comprising 4 sets of 2 paused box squats accommodated with bands alternated with 2 standing broad jumps. The rest period between the squats and the jumps and between the sets was 90 seconds. A control protocol with standing broad jumps only was performed on a separate session. A standing broad jump was performed ∼2 minutes before each protocol and served as a baseline measurement. Standing broad jump distance was significantly greater (4.0 ± 3.4% to 5.7 ± 4.7%) than baseline during the 4 sets of the contrast PAP protocol with the changes being medium in the first, second, and fourth sets (effect size [ES]: 0.58, 0.67, and 0.69, respectively) and large for the third set (ES: 0.81). Conversely, no PAP effect was observed in the control protocol. Additionally, the stronger players displayed a larger PAP effect during each of the 4 sets of the contrast PAP protocol (Cohen's d: 0.28-1.68) and a larger mean effect across these 4 sets (Cohen's d: 1.29). Horizontal jump performance is potentiated after only 90 seconds of rest after an accommodating exercise, and this PAP effect can be elicited across 4 sets. Additionally, the PAP response is largely mediated by the individual's strength level. These results are of great importance for coaches seeking to incorporate PAP complexes involving horizontal jumps in their training programs.

Influence of variable resistance loading on subsequent free weight maximal back squat performance.

The purpose of the study was to determine the potentiating effects of variable resistance (VR) exercise during a warm-up on subsequent free-weight resistance (FWR) maximal squat performance. In the first session, 16 recreationally active men (age = 26.0 ± 7.8 years; height = 1.7 ± 0.2 m; mass = 82.6 ± 12.7 kg) were familiarized with the experimental protocols and tested for 1 repetition maximum (1RM) squat lift. The subjects then visited the laboratory on 2 further occasions under either control or experimental conditions. During these conditions, 2 sets of 3 repetitions of either FWR (control) or VR (experimental) squat lifts at 85% of 1RM were performed; during the experimental condition, 35% of the load was generated from band tension. After a 5-minute rest, 1RM, 3D knee joint kinematics, and vastus medialis, vastus lateralis, rectus femoris, and semitendinosus electromyogram (EMG) signals were recorded simultaneously. No subject increased 1RM after FWR, however, 13 of 16 (81%) subjects increased 1RM after VR (mean = 7.7%; p < 0.01). Lower peak and mean eccentric (16-19%; p ≤ 0.05) and concentric (12-21%; p ≤ 0.05) knee angular velocities were observed during the 1RM following VR when compared with FWR, however, no differences in knee flexion angle (1.8°; p > 0.05) or EMG amplitudes (mean = 5.9%; p > 0.05) occurred. Preconditioning using VR significantly increased 1RM without detectable changes in knee extensor muscle activity or knee flexion angle, although eccentric and concentric velocities were reduced. Thus, VR seems to potentiate the neuromuscular system to enhance subsequent maximal lifting performance. Athletes could thus use VR during warm-up routines to maximize squat performance.

Assessment of the risk factors of coronary heart events based on data mining with decision trees.

Coronary heart disease (CHD) is one of the major causes of disability in adults as well as one of the main causes of death in the developed countries. Although significant progress has been made in the diagnosis and treatment of CHD, further investigation is still needed. The objective of this study was to develop a data-mining system for the assessment of heart event-related risk factors targeting in the reduction of CHD events. The risk factors investigated were: 1) before the event: a) nonmodifiable-age, sex, and family history for premature CHD, b) modifiable-smoking before the event, history of hypertension, and history of diabetes; and 2) after the event: modifiable-smoking after the event, systolic blood pressure, diastolic blood pressure, total cholesterol, high-density lipoprotein, low-density lipoprotein, triglycerides, and glucose. The events investigated were: myocardial infarction (MI), percutaneous coronary intervention (PCI), and coronary artery bypass graft surgery (CABG). A total of 528 cases were collected from the Paphos district in Cyprus, most of them with more than one event. Data-mining analysis was carried out using the C4.5 decision tree algorithm for the aforementioned three events using five different splitting criteria. The most important risk factors, as extracted from the classification rules analysis were: 1) for MI, age, smoking, and history of hypertension; 2) for PCI, family history, history of hypertension, and history of diabetes; and 3) for CABG, age, history of hypertension, and smoking. Most of these risk factors were also extracted by other investigators. The highest percentages of correct classifications achieved were 66%, 75%, and 75% for the MI, PCI, and CABG models, respectively. It is anticipated that data mining could help in the identification of high and low risk subgroups of subjects, a decisive factor for the selection of therapy, i.e., medical or surgical. However, further investigation with larger datasets is still needed.

Evaluation and comparison of fluorescence polarization assay with three of the currently used serological tests in diagnosis of human brucellosis.

Fluorescence polarization assay (FPA) is a method that has been used for the diagnosis of brucellosis in animals for many years. To test its possible usefulness for the diagnosis of human brucellosis, 230 sera from patients with clinical signs of brucellosis and positive serological tests (Rose Bengal, Standard Agglutination Test, iELISA), and 305 sera from a healthy population with no clinical/epidemiological/serological evidence were examined with FPA. By using ROC analysis, the cut-off value was estimated at 99 mP, with 93.5% sensitivity (95% CI 89.5-96.3) and 96.1% specificity (95% CI 93.2-97.9). The pairwise comparison of ROC curves between FPA and iELISA and between FPA and RBT revealed no significant statistic difference (P < 0.05). On the contrary it revealed a significant statistic difference between FPA and SAT (P > 0.05). SAT also had the lowest sensitivity (81.7%) among the three tests used in case definition while iELISA had a sensitivity of 90.8% and RBT a sensitivity of 88.7%. The Kappa analysis showed that FPA has a very good agreement (0.92) with the "status of the disease" and with iELISA (0.837). According to our results, FPA seems to be a valuable method for the diagnosis of brucellosis in humans. Taking into consideration the advantages of the method such as the speed of results obtaining, the objectivity of results interpretation, as well as the cost, FPA could be considered as a replacement for other established methods. However, further studies are needed to assess the reproducibility of FPA.

Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats.

Fluorescence polarization assay (FPA) is a relatively new test for the serological diagnosis of Brucella spp. infection in animals. FPA, carried out in 96-well microplate format, was validated here for diagnosing B. melitensis infection in sheep and goats. This study included sera from 1933 sheep and goats, from animals reared in naturally infected flocks (verified by culture) and showing a positive reaction to two different tests conducted in parallel. In addition, 2154 sera originating from healthy sheep and goats, reared in areas where B. melitensis had never been isolated, were assayed. The optimum cut-off value offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 15 mP over the mean value of the buffer control used in each microplate as determined by receiver operating characteristic analysis. The DSn and DSp of the FPA for small ruminants carried out in microplates at this cut-off value were calculated to be 95.9% and 97.9% with 95% confidence intervals (95% CI) of 94.9-97.7% and 97.2-98.4%, respectively. The accuracy of the FPA, as expressed by determination of the area under the curve, was 0.991. Indirect ELISA and FPA tests offered the highest DSn when compared with the Rose Bengal test, the complement fixation test, the modified Rose Bengal test and competitive ELISA. The parallel or serial combination of FPA with indirect ELISA offers the highest DSn and DSp. As temperature can affect the results of the FPA, all reagents must be at the same temperature and the standard for comparison must always be read under the same conditions as the sera under test. FPA performed in microplates is a promising assay; the DSn and accuracy are better than those of the tests currently approved for diagnosing B. melitensis in small ruminants. Because of its simplicity, speed, and accuracy, this test can improve capacity for laboratory testing and the efficacy of an eradication program based on a test-and-slaughter policy.

Assessment of serological response of young and adult sheep to conjunctival vaccination with Rev-1 vaccine by fluorescence polarization assay (FPA) and other serological tests for B. melitensis.

The serological response of young and adult sheep vaccinated conjunctivally with Rev-1 vaccine was assessed by fluorescence polarization assay (FPA), Rose Bengal test (RBT), complement fixation test (CFT), modified Rose Bengal test (m-RBT), indirect ELISA (i-ELISA) and competitive ELISA (c-ELISA), at different post vaccination intervals. One hundred and thirty six adult sheep and 64 lambs were used in the study. The vaccinated animals were bled prior to vaccination (0 day) and thereafter at 21st, 42nd, 35th, 63rd, 91st, 125th, 159th, and 223rd and 330th day post vaccination. The majority of animals (young and adult) showed positive reaction by FPA, RBT, CFT, m-RBT and c-ELISA 21 days post vaccination, whereas by i-ELISA at 42 days. All tests perform equal when animals vaccinated as young are tested 125 days (4 months) post vaccination. In case of animals vaccinated at adulthood, FPA, RBT, CFT and c-ELISA perform equal if the animals are tested 223 days (approximately 8 months) post vaccination. I-ELISA and m-RBT show low specificity if ewes vaccinated at adulthood are tested 330 days (11 months) post vaccination. If control of brucellosis in sheep is based on conjunctivally vaccination of lambs with Rev-1, the vaccinated animals can be tested by any test used for diagnosis of B.melitensis infection accurately at least 4 months post vaccination. If brucellosis control is based on mass vaccination the use of m-RBT and i-ELISA is not recommended for testing adult animals at least for 330 days (11 months) post vaccination due to tests low specificity. Further research is needed so the appropriate cut-offs to be established for FPA, c-ELISA or i-ELISA to become valuable tools for the eradication of Brucella spp. infection in small ruminants in areas where vaccination is practiced.

Validation of fluorescence polarization assay (FPA) and comparison with other tests used for diagnosis of B. melitensis infection in sheep.

Fluorescence polarization assay (FPA) is a new test for the serological diagnosis of Brucella spp. infection in animals. The FPA is validated for the diagnosis of B. melitensis infection in sheep. For this purpose, 166 sera originated from natural infected sheep (verified by culture) and 851 sera originated from healthy animals (reared in areas where B. melitensis was never been isolated) were tested. The optimum cut-off value that offers the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 87mP with the use of ROC analysis. The DSn and DSp of FPA using this cut-off value are calculated at 97.6 and 98.9% with a 95% confidence interval (CI) of 93.9-99.3% and 98.0-99.5%, respectively. The DSn and DSp of FPA have been assessed also using as positive reference (n=587), sera that gave positive results at least in two tests used for diagnosis of B. melitensis in sheep as Rose Bengal Test (RBT), modified Rose Bengal Test (m-RBT), complement fixation test (CFT), indirect Elisa (i-Elisa) and competition Elisa (c-Elisa) originated from animals reared in flocks infected by B. melitensis. The optimum cut-off value using the above panel of positive reference sera was the same offering a DSn of 95.9% with a 95% CI, 94.0-97.4%, since the DSp remains the same. The DSn and DSp as well as performance, accuracy and agreement of FPA's result were compared with those of other tests used. The accuracy of FPA is very high, similar with that of i-Elisa. FPA is a promising assay, which offers a DSn and accuracy better that of those of the tests currently approved for the diagnosis of B. melitensis in sheep and goats. Due to its simplicity, the sort time that results can be obtained and its accuracy it can be used and improve the laboratory testing capacity as well as the efficacy of the eradication program based on test-and-slaughter policy.

Epidemiology of dermatomycoses in children living in Northern Greece 1996-2000.

In the 5-years period, 1996-2000, 1045 children under 13 years old were examined for suspected dermatomycosis. In 611 cases fungi were isolated. Male children were mainly affected on the scalp and body area. Girls were more affected in the location of the arms and legs. There was a greater proportion of cases in the age range 2-12 years. The most prominent fungus was Microsporum canis (515 cases) followed by Trichophyton rubrum (34) and Candida albicans (20). Tinea capitis (280 cases) mainly caused by M. canis (276 cases) was the most common clinical form. Tinea corporis (109 cases) mainly caused by M. canis (88 cases), C. albicans (10 cases) and T. rubrum (seven cases) was the second most frequent clinical form.

Towards single screening tests for brucellosis.

This paper describes an indirect enzyme-linked immunosorbent assay (I-ELISA) and a fluorescence polarisation assay (FPA), each capable of detecting antibody in several species of hosts to smooth and rough members of the genus Brucella. The I-ELISA uses a mixture of smooth lipopolysaccharide (SLPS) and rough lipopolysaccharide (RLPS) as the antigen, and a recombinant protein A/G conjugated with horseradish peroxidase as the detection reagent. When using individually determined cutoff values, the SLPS/RLPS combined-antigen I-ELISA detected antibody in slightly more animals exposed to SLPS or to RLPS than did I-ELISA procedures using each individual antigen separately. Similarly, the assay using combined antigens detected antibody in slightly fewer animals not exposed to Brucella sp. When a universal cutoff of 10% positivity was used (relative to strongly positive control sera of each species), the overall performance index (percentage sensitivity plus percentage specificity) value decreased by 1.0 (from 199.4 to 198.4). In the FPA, it was not possible to use a universal cutoff without significant loss of performance. The overall sensitivity value for the FPA using the combined FPA antigen was 1.0% lower than using the O-polysaccharide (OPS) from SLPS and 9.1% higher than using the core antigen (CORE) from RLPS. When the combined antigen was used, the FPA specificity was slightly higher (1.2%) than from only the OPS, and considerably higher (12.6%) than the CORE. Overall, both the I-ELISA and the FPA with combined antigens were suitable as screening tests for all species of Brucella in the animal species tested.