Evaluation of new instruments for screening and diagnosis of tics and tic disorders in a well-characterized sample of youth with tics and recruited controls.

Tics and tic disorders can significantly impact children, but limited screening tools and diagnostic challenges may delay access to care. The current study attempted to address these gaps by evaluating sensitivity and specificity of the Motor or Vocal Inventory of Tics (MOVeIT), a tic symptom screener, and the Description of Tic Symptoms (DoTS), a brief diagnostic assessment for tic disorders. Children (n=100, age 6-17 years old) with tic disorders attending a Tourette specialty clinic and a community-recruited sample without tics completed a gold-standard assessment by a tic expert; these evaluations were compared to child self-report and parent and teacher report versions of the MOVeIT, and child and parent versions of the DoTS. The parent and child MOVeIT met or exceeded pre-specified 85% sensitivity and specificity criteria for detecting the presence of tics when compared to a gold-standard tic expert diagnosis. The Teacher MOVeIT had lower sensitivity (71.4%) but good specificity (95.7%) for identifying any tic symptoms compared to gold standard. For determination of the presence or absence of any tic disorder, sensitivity of both parent and child DoTS was 100%; specificity of the parent DoTS was 92.7% and child DoTS specificity was 75.9%. More work may be needed to refine the teacher MOVeIT, but it is also recognized that tic expression may vary by setting. While the MOVeIT and DoTS parent and child questionnaires demonstrated adequate sensitivity and specificity for determining the presence of tics and tic disorders in this well-defined sample, additional testing in a general population is warranted.

Impact of the induction phase chemotherapy on cytokines and oxidative markers in peripheral and bone marrow plasma of children with acute lymphocytic leukemia.

B-cell acute lymphocytic leukemia (B-ALL) is the main neoplasia affecting children worldwide, in which cytotoxic chemotherapy remains the main treatment modality. In this study, we analyzed the profile of inflammatory markers concerning oxidative stress and cytokines in 17 B-ALL patients. Peripheral blood (PB) and bone marrow (BM) samples were collected and evaluated for the pro-oxidative status (nitric oxide products-NOx and hydroperoxides), antioxidants (sulfhydryl groups-SH and total radical-trapping antioxidant parameter-TRAP), and cytokines (TNF-α, IFN-γ), at diagnosis (D0) to and the end of the induction phase (D28). At D28, hydroperoxides were higher in PB, concomitant to TNF-α levels. INF-γ was increased in the BM at D28. Hydroperoxides were higher in patients presenting malignant cells in BM and/or PB after treatment, a condition named minimal residual disease (MRD) when compared to those without MRD at D28. These findings suggest that oxidative stress and cytokines vary across the B-ALL induction phase, and lipid peroxidation is a potential marker associated with MRD status.

¿Está el estrés oxidativo asociado a la gravedad de la enfermedad, a la función pulmonar y al síndrome metabólico en la enfermedad pulmonar obstructiva crónica? / Is oxidative stress associated with disease severity, pulmonary function and metabolic syndrome in chronic obstructive pulmonary disease?

OBJETIVO: Investigar las asociaciones entre los biomarcadores oxidantes/antioxidantes y el estado de gravedad, la función pulmonar y la presencia de síndrome metabólico (SM) en pacientes con EPOC. MÉTODOS: Se incluyeron 74 sujetos, 39 con EPOC (edad 69+/-7 años; mujeres 41%) y 35 para el grupo control (edad 69+/-7 años; mujeres: 43%). Fueron diagnosticados con SM y asignados a uno de los 4 subgrupos: EPOC y control, con y sin SM, respectivamente. Se analizaron los productos de oxidación avanzada de proteína (AOPP), la paraoxonasa-1, la actividad de catalasa, el grupo sulfhidrilo y el hidroperóxido de lípidos totales. La función pulmonar fue analizada por medio de un pletismógrafo. RESULTADOS: El estado de gravedad de la EPOC (GOLD≥3) y la función pulmonar fueron asociados con el grupo sulfhidrilo y AOPP (p≤0,03 para todos). La prevalencia de SM se asoció con AOPP en la EPOC (p = 0,04). Los individuos con EPOC y SM mostraron niveles de AOPP más altos en comparación con los sujetos con EPOC sin SM (p < 0,0001). CONCLUSIÓN: La gravedad de la EPOC, el deterioro de la función pulmonar y la presencia de síndrome metabólico están asociados con el estrés oxidativo en individuos con EPOC

OBJECTIVE: To investigate associations between oxidant/antioxidant biomarkers with the disease severity, pulmonary function and diagnosis of metabolic syndrome (MetS) in patients with COPD. METHODS: Seventy-four subjects were included, 39 with COPD (age 69+/-7 years; female 41%) and 35 for control group (age 69+/-7 years; female 43%). They were diagnosed with MetS and allocated in one of 4 subgroups: COPD and control, with and without MetS, respectively. Advanced oxidation protein products (AOPP), paraoxonase-1, catalase activity, sulfhydryl group and total lipid hydroperoxide were assayed. Pulmonary function was performed with a plethysmograph. RESULTS: COPD severity (GOLD≥3) and pulmonary function were associated with sulfhydryl group and AOPP (P≤.03 for all). The prevalence of MetS was associated with AOPP in COPD (P=.04). Individuals with COPD and MetS showed higher AOPP compared to COPD without MetS (P<.0001). CONCLUSION: COPD severity, worse pulmonary function and presence of metabolic syndrome are associated with oxidative stress in individuals with COPD

Is oxidative stress associated with disease severity, pulmonary function and metabolic syndrome in chronic obstructive pulmonary disease? / ¿Está el estrés oxidativo asociado a la gravedad de la enfermedad, a la función pulmonar y al síndrome metabólico en la enfermedad pulmonar obstructiva crónica?

OBJECTIVE: To investigate associations between oxidant/antioxidant biomarkers with the disease severity, pulmonary function and diagnosis of metabolic syndrome (MetS) in patients with COPD. METHODS: Seventy-four subjects were included, 39 with COPD (age 69±7 years; female 41%) and 35 for control group (age 69±7 years; female 43%). They were diagnosed with MetS and allocated in one of 4 subgroups: COPD and control, with and without MetS, respectively. Advanced oxidation protein products (AOPP), paraoxonase-1, catalase activity, sulfhydryl group and total lipid hydroperoxide were assayed. Pulmonary function was performed with a plethysmograph. RESULTS: COPD severity (GOLD≥3) and pulmonary function were associated with sulfhydryl group and AOPP (P≤.03 for all). The prevalence of MetS was associated with AOPP in COPD (P=.04). Individuals with COPD and MetS showed higher AOPP compared to COPD without MetS (P<.0001). CONCLUSION: COPD severity, worse pulmonary function and presence of metabolic syndrome are associated with oxidative stress in individuals with COPD.

Does calretinin immunohistochemistry reduce inconclusive diagnosis in rectal biopsies for Hirschsprung disease?

Many difficulties occur during the evaluation of rectal biopsies for the diagnosis of Hirschsprung disease. We investigated whether the introduction of calretinin (CR) immunohistochemistry in a diagnostic panel could decrease the rate of inconclusive results. Data from 82 patients undergoing rectal biopsies before and after CR introduction were analyzed. Inconclusive results were obtained in 17 of 45 rectal biopsies (37.8%) in the series of cases before CR introduction and in 5 of 42 rectal biopsies (11.9%) in the series of cases after CR (P < 0.006). The inclusion of CR in the histopathologic panel may improve the diagnostic accuracy of Hirschsprung disease.

Accuracy of the iPex multi-frequency electronic apex locator in primary molars: an ex vivo study.

AIM: To evaluate ex vivo the accuracy of the iPex multi-frequency electronic apex locator (NSK Ltd, Tokyo, Japan) for working length determination in primary molar teeth. METHODOLOGY: One calibrated examiner determined the working length in 20 primary molar teeth (total of 33 root canals). Working length was measured both visually, with the placement of a K-file 1 mm short of the apical foramen or the most coronal limit of root resorption, and electronically using the electronic apex locator iPex, according to the manufacturers' instructions. Data were analysed statistically using the intraclass correlation (ICC) test. RESULTS: Comparison of the actual and the electronic measurements revealed high correlation (ICC = 0.99) between the methods, regardless of the presence or absence of physiological root resorption. CONCLUSIONS: In this laboratory study, the iPex accurately identified the apical foramen or the apical opening location for working length measurement in primary molar teeth.

Graft versus host disease in a rat small bowel transplant model after T-cell depleted donor specific bone marrow infusion

Low cytoreductive regimen of irradiation associated to unmodified bone marrow infusion (UBM) does not prevent the occurrence of graft versus host disease (GVHD) after transplant. PURPOSE: In this study we evaluated the potential advantages of a long-term immunossupression and T-cell depleted bone marrow infusion (TCDBMI) in preventing the occurrence of GVHD after small bowel transplantation (SBTx). METHODS: Heterotopic SBTX was performed with Lewis rats as recipients and DA as donors and distributed into 5 groups according to the irradiation, duration of immunossupression and the use of UBM or TCDBMI: G1 (n=6), without irradiation and G2 (n=9), G3 (n=4), G4 (n=5) and G5 (n=6) was given 250 rd of irradiation. Groups 1,2,4 and G3 and 5 were infused with 100 x 106 UBM and TCDBM respectively. Animals in G1, 2, 3 were immunossupressed with 1mg/ FK506/Kg/IM for 5 days and G4 and G5 for 15 days. Anti CD3 monoclonal antibodies and immunomagnetic beads were used for T-cell depletion.Animals were examined for rejection, GVHD, chimerism characterization and ileal and skin biopsies. RESULTS: Minimal to mild rejection was observed in all groups; however, GVHD were present only in irradiated groups. Long-term immunossupression changed the severity of GVHD in G4 and G5. Rejection was the cause of death in G1 while GVHD in G2, 3, 4 and 5, not avoided by the use of TCDBMI. Total chimerism and T-cell chimerism was statistically higher in irradiated groups when compared to G1. CONCLUSION: Extended immunossupression associated to low dose of irradiation decrease the severity of GVHD, not avoided by the use of TCDBMI.

Chimerism induction by nonmyeloablactive preconditioning and bone marrow infusion in rat small bowel transplantation

ABSTRACT: In our previous work we demonstrated that the use of donor specific bone marrow infusions ( DSBMI ) after small bowel transplantation did not improve the graft survival after a short course of immunossupression. PURPOSE: In the current study, we evaluated whether recipient preconditioning with different regimens of radiation combined with DSBMI may enhance small bowel allograft survival with minimum recipient morbidity. METHODS: Heterotopic small bowel transplantation (SBTx) was performed with Lewis rats as recipients and DA rats as donors, which were immunossupressed with a short course of tacrolimus (FK 506 ) at 1mg/Kg/day for 5 days and distributed in 4 groups: group 1 (n= 4) without both irradiation and DSBMI; Groups 2 (n= 6), 3 (n= 9) and 4 (n= 6) received 100 x 106 DSBM cells at the time of the transplant. Groups 3 and 4 were irradiated with 250 and 400 rd respectively. Animals were examined daily for clinical signs of rejection or GVHD. Blood samples were taken weekly for chimeric studies by FC and intestinal biopsies were performed every 2 weeks. RESULTS: Animals in G1 and G2 had minimal rejection at day 15 after SBTx while GVHD was clinically and histologically characterized in G 3 and G 4. Total chimerism and T-cell chimerism was higher in irradiated groups when compared to non-irradiated groups. With exception of G1 and 2 where rejection was the cause of death, all animals in G3 and 4 died of GVHD. CONCLUSION:We concluded that low cytoreductive of irradiation can successfully decrease the graft rejection but not prevent the occurrence of GVHD.

The use of ultrasound for cleaning the surface of stainless steel and nickel-titanium endodontic instruments.

AIM: The aim of this study was to evaluate the efficacy of ultrasound in cleaning the surface of stainless steel and Ni-Ti endodontic instruments. METHODOLOGY: Twenty nickel-titanium instruments (10 Quantec files and 10 Nitiflex) and 20 stainless steel K-files (10 Maillefer-Dentsply and 10 Moyco Union Broach) were removed from their original packages and evaluated using a scanning electron microscope. Scores were given for the presence of residues on the surface of the instruments. The instruments were then cleaned in an ultrasonic bath containing only distilled water or detergent solution for 15 min, and re-evaluated, using scanning electron microscopy. RESULTS: Before cleaning, a greater amount of metallic debris was observed on the nickel-titanium Quantec instruments (P < 0.05), when compared to those made of stainless steel. Statistical analysis showed that the use of ultrasound was effective for cleaning the instruments, regardless of the irrigating solution or the instruments type (P < 0.05). CONCLUSIONS: The use of ultrasound proved to be an efficient method for the removal of metallic particles from the surface of stainless steel and Ni-Ti endodontic instruments.

In vitro evaluation of the antimicrobial activity of a castor oil-based irrigant.

The antimicrobial activity of irrigating solutions--Endoquil (castor oil detergent), 2% chlorhexidine gluconate solution, and 0.5% NaOCl solution-was evaluated against gram-positive cocci (Micrococcus luteus, Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, Streptococcus mutans, and Streptococcus sobrinus), gram-negative rods (Escherichia coli and Pseudomonas aeruginosa), and the yeast Candida albicans. Activity was evaluated using the two-layer agar diffusion technique. The base layer was obtained by pouring 10.0 ml of Muller Hinton Medium or 10.0 ml of Brain Heart Infusion agar in a Petri dish. After solidification a 5.0 ml seed layer of Muller Hinton Medium or Brain Heart Infusion agar with inoculum (106/ml) was added. Absorbent paper disks (6.0 mm in diameter) immersed in the solutions were placed at equidistant points. Plates were maintained at room temperature for 2 h for prediffusion of the solutions and incubated at 37 degrees C for 24 h. The candle jar system was used for the Brain Heart Infusion agar plates. All tests were performed in duplicate. After incubation the medium was optimized with 0.05 g% triphenyltetrazolium chlorate gel and inhibition halos were measured. All bacterial strains were inhibited by 2.0% chlorhexidine gluconate. Endoquil was effective against gram-positive microorganisms, and 0.5% NaOCl was effective only against S. aureus.

In vitro evaluation of antimicrobial activity of sealers and pastes used in endodontics.

The antimicrobial activity of four root canal sealers (AH Plus, Sealapex, Ketac Endo, and Fill Canal), two calcium hydroxide pastes (Calen and Calasept), and a zinc oxide paste was evaluated. Seven bacterial strains were used, six of them standard; Micrococcus luteus ATCC 9341, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Staphylococcus epidermidis ATCC 12228, Escherichia coli ATCC 25922, and Enterococcus faecalis ATCC 10541. There was a wild strain of Streptococcus mutans isolated from saliva obtained in an adult dental clinic. Activity was evaluated using the agar diffusion method with Brain Heart Infusion agar and Müller Hinton medium seeded by pour plate. Calcium hydroxide-based sealers and pastes were either placed directly into 4.0 x 4.0 mm wells or by using absorbent paper points. The plates were kept at room temperature for 2 hr for diffusion. After incubation at 37 degrees C for 24 hr, the medium was optimized with 0.05 g% TTC gel and inhibition haloes were measured. All bacterial strains were inhibited by all materials using the well method. However, when the materials were applied with absorbent paper points, Enterococcus faecalis was not inhibited by zinc oxide, and Pseudomonas aeruginosa was not inhibited by AH Plus, Fill Canal, and the zinc oxide-based paste. We conclude that sealers and pastes presented antimicrobial activity in vitro and culture medium optimization with 0.05 g% TTC gel facilitated observation of the inhibition haloes.

In vivo antimicrobial activity of 2% chlorhexidine used as a root canal irrigating solution.

The aim of the present study was to evaluate the in vivo antimicrobial activity of 2% chlorhexidine gluconate (FCFRP-USP) used as a root canal irrigating solution in teeth with pulp necrosis and radiographically visible chronic periapical reactions. Culture techniques and measurement of the inhibition zone were used. Twenty-two root canals of incisors and molars of 12 patients were used. After accessing the canal, the first root canal sample was collected with two sterile paper points that were transferred to a tube containing reduced transport fluid. The root canal was instrumented using chlorhexidine solution. A small sterile cotton pellet was placed at the root canal entrance, and the cavity was sealed with zinc oxide-eugenol cement. The canals were maintained empty for 48 h. Three sterile paper points were then introduced to absorb the root canal fluid (second sample). One paper point was placed on an agar plate inoculated with Micrococcus luteus ATCC 9341 and incubated for 24 h at 37 degrees C, and the other two were submitted to microbiological evaluation. Present in 10 cases at baseline, mutans streptococci was reduced by 100% at the second assessment. Treatment showed an efficiency of 77.78% for anaerobic microorganisms at the second assessment. These data suggest that chlorhexidine prevents microbial activity in vivo with residual effects in the root canal system up to 48 h.

Evaluation of the antimicrobial activity of three irrigating solutions in teeth with pulpal necrosis.

The antimicrobial activity of 0.4% papaine gel (FCF-USP), an antibacterial product derived from 3.3% castor oil (IQSC-USP), and 0.5% sodium hypochlorite (FORP-USP) was evaluated in teeth with radiographically visible pulpal necrosis and periapical lesion in vivo. After cavity access, under aseptic conditions, a first harvesting was performed. The 3 irrigating solutions were used for biomechanical preparation. After 72 hours, a second harvesting was performed, also under aseptic conditions. The number of colony forming units (cfu) was counted with a stereomicroscope under reflected light. Castor oil and 0.5% sodium hypochlorite presented similar antimicrobial activities for the reduction of the anaerobe number, S. mutans and streptococci; however, the papaine gel showed lower activity. We conclude that both castor oil and sodium hypochlorite are effective as antimicrobial agents and can be used in the treatment of root canals with pulpal necrosis.

Evaluation of chemiluminescence, procoagulant activity and antigen presentation by monocytes from lepromatous leprosy patients with or without reactional episodes.

In this study, we evaluated the activity of peripheral blood mononuclear cells (PBMC), isolated from treated and untreated lepromatous leprosy patients, from lepromatous leprosy patients during and after reactional episodes (erythema nodosum leprosum (ENL) and reversal reaction (RR)), and from normal healthy individuals. We determined reactive oxygen intermediate (ROI) production, procoagulant activity (PCA) and HLA-DR antigen expression of monocytes, besides lymphoproliferation, both in the presence and absence of various stimulatory agents. Phorbol myristate acetate (PMA) stimulated ROI production by monocytes from all the groups studied, with patients during reactional episodes (ENL and RR) showing a significantly higher response (p < 0.009 and p < 0.00001). Irradiated Mycobacterium leprae, although having little effect when added alone, strongly suppressed PMA-stimulated ROI production. Muramyl dipeptide (MDP) had no influence on either basal or on PMA-induced ROI production. Basal monocyte PCA, as well as M. leprae or concanavalin A (ConA)-induced monocyte PCA was comparable in monocytes from all the groups studied. ConA was able to induce mitogenic activity in mononuclear cells isolated from all the groups studied. M. leprae, although stimulatory for normal individuals, did not induce lymphoproliferation in lepromatous leprosy patients, except for cells from patients during RR, which responded equally to M. leprae and to ConA. The absence of M. leprae-induced lymphoproliferation in lepromatous leprosy patients is not caused by the lack of basal HLA-DR expression, as PBMC from all individuals studied showed the same level of this antigen. Our results suggest an increase of spontaneous or PMA-induced monocyte activity, as detected by ROI production, during the reactional episode; addition of M. leprae suppressed this response. The increase in monocyte activity could be correlated with the increase of lymphoproliferation response to M. leprae during RR, but not during ENL. The importance of a possible immune suppressive action of M. leprae is discussed.

In vitro tumor necrosis factor production by mononuclear cells from lepromatous leprosy patients and from patients with erythema nodosum leprosum.

The production of tumor necrosis factor (TNF) by Mycobacterium leprae-stimulated phagocyte cells, isolated from lepromatous leprosy patients (LL) and normal individuals, was evaluated, using the highly TNF-sensitive mouse fibrosarcoma cell line WEHI164cl13. Mononuclear cells, isolated from all individuals studied, showed a low level of spontaneous TNF production, except for patients undergoing erythema nodosum leprosum (ENL), in which we found significantly higher levels of TNF. Addition of M. leprae to the phagocyte cell culture enhanced TNF production in all groups studied, except in the group with untreated leprosy patients. Strongest M. leprae-induced TNF release was found in mononuclear cell cultures derived from ENL patients. Patients in the postreactional state showed significantly higher TNF levels than healthy controls. These findings support the idea that TNF plays a key role in the complex symptomatology of ENL.