Abstract This is a nonclinical, controlled, and triple-blind study to investigate the effects of codeine-associated geraniol on the modulation of orofacial nociception and its potential central nervous system depressing effect in an animal model. The orofacial antinociceptive activity of geraniol in combination with codeine was assessed through the following tests: (i) formalin-induced pain, (ii) glutamate-induced pain, and (iii) capsaicin-induced pain. Six animals were equally distributed into six groups and received the following treatments, given intraperitoneally (i.p.) 30 minutes before the experiments: a) geraniol/codeine 50/30 mg/kg; b) geraniol/codeine 50/15 mg/kg; c) geraniol/codeine 50/7.5 mg/kg; d) geraniol 50 mg/kg; e) codeine 30 mg/kg (positive control); or f) 0.9% sodium chloride (negative control). We performed pain behavior analysis after the injection of formalin (20 µL, 20%), glutamate (20 µL, 25 µM), and capsaicin (20 µL, 2.5 µg) into the paranasal region. Rubbing time of the paranasal region by the hind or front paw was used as a parameter. In the neurogenic phase of the formalin test, the geraniol/codeine at 50/7.5 mg/kg was able to promote the maximum antinociceptive effect, reducing nociception by 71.9% (p < 0.0001). In the inflammatory phase of the formalin test, geraniol/codeine at 50/30 mg/kg significantly reduced orofacial nociception (p < 0.005). In the glutamate test, geraniol/codeine at 50/30 mg/kg reduced the rubbing time by 54.2% and reduced nociception in the capsaicin test by 66.7% (p < 0.005). Geraniol alone or in combination does not promote nonspecific depressing effects on the central nervous system. Based on our findings, we suggest the possible synergy between geraniol and codeine in the modulation of orofacial pain. nonclinical controlled tripleblind triple blind codeineassociated associated model tests i (i formalininduced induced ii (ii glutamateinduced iii (iii capsaicininduced treatments i.p. ip p (i.p. 3 experiments geraniolcodeine 5030 50/3 mgkg mg kg b 5015 15 50/1 c 5075 7 5 50/7. d e positive control control) f 09 0 9 0.9 negative control. . 20 (2 µL 20%, 20% , 20%) 2 µM, µM µM) 2. µg parameter 719 71 71.9 0.0001. 00001 0.0001 0001 0.0001) 0.005. 0005 0.005 005 0.005) 542 54 54.2 667 66 66.7 findings i.p (i.p 503 50/ 501 1 507 50/7 0. ( 71. 0000 0.000 000 0.00 00 54. 6 66. 0.0

Resumo O objetivo deste trabalho foi investigar a influência do estágio reprodutivo da cevada de florescimento fechado na infecção por Fusarium graminearum e avaliar a eficácia da aplicação pré-inoculação de fungicida no controle da fusariose da espiga e na redução da contaminação dos grãos pela micotoxina desoxinivalenol (DON). Dois experimentos, em delineamento completamente casualizado, com oito repetições, foram conduzidos em câmara de crescimento com diferentes temperaturas. A análise estatística foi não paramétrica, tendo-se utilizado o teste de Kruskal-Wallis. Plantas da cultivar BRS BRAU de cevada, em vasos, foram inoculadas por pulverização de suspensão de conídios, nos seguintes três estágios reprodutivos: metade da antese, grão leitoso e grão massa mole. O fungicida tebuconazol foi pulverizado 48 horas antes da inoculação do patógeno. Os sintomas de fusariose da espiga causados pela inoculação de F. graminearum no estágio de grão leitoso são claramente visíveis, mas são leves nos estágios de metade da antese e grão com massa mole. A inoculação nos três estágios resulta em contaminação dos grãos com DON, que é reduzida com a aplicação de fungicida pré-inoculação. préinoculação pré DON. DON . (DON) experimentos casualizado repetições temperaturas paramétrica tendose tendo se KruskalWallis. KruskalWallis Kruskal Wallis. Wallis Kruskal-Wallis vasos conídios reprodutivos mole 4 patógeno F visíveis préinoculação. inoculação. (DON

Abstract The objective of this work was to investigate the influence of the reproductive stage of closed-flowering barley on Fusarium graminearum infection and to evaluate the efficacy of pre-inoculation fungicide application in controlling Fusarium head blight (FHB) and reducing grain contamination with the deoxynivalenol (DON) mycotoxin. Two experiments, in a completely randomized design with eight replicates, were conducted in a growth chamber at different temperatures. The statistical analysis was nonparametric, using the Kruskal-Wallis test. Potted plants of the BRS BRAU barley cultivar were inoculated by spraying a conidial suspension at the following three reproductive stages: anthesis halfway, grain at medium milk, and soft dough. The tebuconazole fungicide was sprayed 48 hours before pathogen inoculation. The FHB symptoms caused by the inoculation with F. graminearum at the medium milk stage are fully visible, although fungicide application before pathogen inoculation controls the disease, but mild at the anthesis halfway and soft dough stages. Inoculation at the three stages results in grain contamination with DON, which is reduced by pre-inoculation fungicide application. closedflowering closed flowering preinoculation pre (FHB DON (DON mycotoxin experiments replicates temperatures nonparametric KruskalWallis Kruskal Wallis test 4 F visible disease

Abstract Betamethasone (BET) is a synthetic glucocorticoid recommended for pregnant women at imminent risk of preterm birth before 34 weeks to reduce neonatal complications. There are different techniques to describe BET plasma quantification. However, none quantified the plasmatic concentration of BET in dichorionic (DC) twin pregnancies using LC-MS. Our objectives were to develop and validate a method for quantifying BET by LC-MS for pharmacokinetic (PK) and placental transfer studies in DC twin pregnancies. Blood samples were collected after intramuscular administration of a single BET dose containing 6 mg disodium phosphate + 6 mg acetate. BET was determined in plasma by liquid-liquid extraction. The method showed linearity in the range of 2-250 ng/mL, as well as precision and accuracy with a coefficient of variation and relative standard errors ≤ 15%. Additionally, the method presented selectivity and did not present matrix or carry-over effect. Stability tests also presented coefficient of variation and relative standard errors ≤ 15%. This is the first study which describe maternal and fetal plasma concentrations of BET in a DC twin pregnancy. The BET PK parameters were AUC0-∞, CL/F, Vd/F, Cmax, Tmax of 292.20 h*ng/mL, 39.08 L/h, 278.72 L, 25.55 ng/mL and 0.58 h, respectively. The placental transfer ratios of umbilical vein/maternal vein and intervillous space/maternal vein were 0.14 and 0.19 and 0.40 and 0.27 for both twins, respectively. However, a clinical study with more subjects is imperative to confirm this higher concentration of BET in the intervillous space. (BET 3 complications quantification However (DC LCMS. LCMS LC MS. MS (PK acetate liquidliquid liquid extraction 2250 2 250 2-25 ngmL ng mL 15 15% Additionally carryover carry over effect pregnancy AUC0∞, AUC0 AUC ∞, AUC0-∞ CLF CL F CL/F VdF Vd Vd/F Cmax 29220 292 20 292.2 hngmL h h*ng/mL 3908 39 08 39.0 Lh L L/h 27872 278 72 278.7 2555 25 55 25.5 058 0 58 0.5 respectively veinmaternal spacematernal space 014 14 0.1 019 19 040 40 0.4 027 27 0.2 twins 225 2-2 1 AUC0∞ ∞ AUC0- 2922 29 292. 390 39. 2787 7 278. 255 5 25. 05 0. 01 04 4 02 22 2-

Abstract Fetal programming suggests that maternal stimulation and nutrition during the period of fetal development can program the progeny. Conjugated linoleic acid (CLA), an isomer of linoleic acid, has been characterized in several aspects, but few studies have been performed on its involvement in reproduction and fetal programming. The aim of this study was to evaluate the F1, F2 and F3 progeny of female mice supplemented with CLA during the pregestational and gestational periods with respect to biometric and reproductive parameters, as well as ovarian morphophysiology. The F1 progeny of mothers supplemented with CLA exhibited stable weight gain, while the F2 progeny showed no effects (P=0.0187 and P=0.0245, respectively). A reduction in Lee's Index was observed in both generations at the second post-weaning evaluation week in the animals treated with CLA (P=0.0100 and P=0.0078, respectively). The F2 generation showed an increase in the anogenital index in both sexes of the animals treated with CLA (P= 0.0114 and P<0.0001, female and male respectively). CLA administration to mothers did not affect any of the following in their progeny: ovarian follicle mobilization (P>0.05), follicle number (P>0.05) and the integrated density of the lipid content of oocytes included in antral follicles (P>0.05). This study evaluated the use of CLA in mothers and found that it did not affect the progeny regarding murine reproductive performance, suggesting that this supplement can be used safely. CLA, , (CLA) aspects F parameters morphophysiology gain P=0.0187 P00187 P 0 0187 (P=0.018 P00245 0245 P=0.0245 respectively. respectively . respectively) Lees Lee s postweaning post weaning P=0.0100 P00100 0100 (P=0.010 P00078 0078 P=0.0078 P= (P 00114 0114 0.011 P00001 0001 P<0.0001 P>0.05, P005 P>0.05 05 (P>0.05 P>0.05. performance safely (CLA P=0.018 P0018 018 (P=0.01 P0024 024 P=0.024 P=0.010 P0010 010 P0007 007 P=0.007 0011 011 0.01 P0000 000 P<0.000 P00 P>0.0 (P>0.0 P=0.01 P001 01 (P=0.0 P002 02 P=0.02 P000 00 P=0.00 001 0.0 P<0.00 P0 P>0. (P>0. P=0.0 (P=0. 0. P<0.0 P>0 (P>0 P=0. (P=0 P<0. P> (P> P=0 P<0 P<

Abstract Objective: To evaluate the efficacy of xylitol varnishes in the inhibition of enamel demineralization in vitro. Material and Methods: Bovine enamel blocks (n=120) were randomly allocated to four groups (n = 30), and the surface hardness (SH) was measured at baseline. The blocks were treated with the following varnishes: 20% xylitol, 20% xylitol plus F (5% NaF), Duraphat™ (5% NaF, positive control), and placebo (no-F/xylitol, negative control). The varnishes were applied and removed after 6 h of immersion in artificial saliva. The blocks were subjected to pH cycles (demineralization and remineralization for 2 and 22h/day, respectively, for 8 days). Surface and cross-sectional hardnesses were measured to calculate the percentage of SH loss (%SHL) and the integrated loss of the subsurface hardness (ΔKHN). Data were statistically analyzed using Kruskal-Wallis and Tukey’s tests (p<0.05). Results: %SHL was significantly decreased by 20% xylitol plus F, Duraphat™, and 20% xylitol varnishes compared to placebo. The use of 20% xylitol plus F varnish led to a significantly lower percentage of SH loss compared to the use of 20% xylitol varnish without F. However, the experimental and commercial varnishes led to significantly lower subsurface demineralization compared to placebo and did not differ from each other. Conclusion: Xylitol varnishes, especially when combined with F, effectively prevent enamel demineralization.

SUMMARY: Mesenchymal stem cells are characterized by in vitro high proliferation and multilineage potential maintenance. This study aimed to isolate and characterize equine YS mesenchymal stem cells and compare these with amniotic membranes. The yolk sac (YS) and amniotic membranes (AM) were obtained from 20 pregnant mares with gestational age around 30 days. Cells were cultured in α-MEM supplemented with 15 % FBS, 1 % antibiotic solution, 1 % L-glutamine and 1 % nonessential amino acids. To cell characterization we used cytogenetic analysis, fibroblast colony-forming unit assays, cell growth curves, immunophenotyping, flow cytometry, differentiation assays and teratoma formation. Results: Both cell sources presented fibroblastoid and epithelioid-like format. The YS cells have lower colony formation potential then AM ones, 3 versus 8 colonies per 103 plated cells. However, YS cells grew progressively while AM cells showed steady. Both, the YS and amnion cells immunolabeled for Oct-4, Nanog, SSEA-3, cytokeratin 18, PCNA, and vimentin. In addition, presented mesenchymal, hematopoietic, endothelial and pluripotency markers in flow cytometry. Discussion: Both cell sources presented high plasticity and differed into osteogenic, adipogenic, and chondrogenic lineages, and no tumor formation in nude mice was observed. The results suggest that horse YS may be useful for cell therapy such as amnion-derived cells.

RESUMEN: Las células madre mesenquimales se caracterizan por una alta proliferación in vitro y un mantenimiento potencial de múltiples líneas. Este estudio tuvo como objetivo aislar y caracterizar las células madre mesenquimales del saco vitelino equinas y compararlas con las membranas amnióticas. Se obtuvo el saco vitelino (SV) y las membranas amnióticas (MA) de 20 yeguas preñadas con edad gestacional de aproximadamente 30 días. Las células se cultivaron en α -MEM suplementado con 15 % de FBS, 1 % de solución antibiótica, 1 % de L-glutamina y 1 % de aminoácidos no esenciales. Para la caracterización celular utilizamos análisis citogenéticos, ensayos de unidades de colonias de fibroblastos, curvas de crecimiento celular, inmunofenotipaje, citometría de flujo, ensayos de diferenciación y formación de teratomas. Ambas fuentes celulares presentaron formato fibroblastoideo y epitelioide. Las células SV tienen un potencial de formación de colonias más bajo que las de MA, 3 versus 8 colonias por 103 células en placa. Sin embargo, las células SV crecieron progresivamente mientras que las células MA se mostraron estables. Tanto las células YS como las células amnios están inmunomarcadas para Oct-4, Nanog, SSEA-3, citoqueratina 18, PCNA y vimentina. Además, presentó marcadores mesenquimales, hematopoyéticos, endoteliales y pluripotenciales en citometría de flujo. Ambas fuentes celulares presentaron alta plasticidad y diferían en linajes osteogénicos, adipogénicos y condrogénicos, y no se observó formación de tumores en ratones. Los resultados sugieren que el SV de caballo puede ser útil para la terapia celular, como las células derivadas de amnios.

ABSTRACT Soils with a long-term history of animal manure application exhibit higher residual effects of nitrogen (N) in soil and can affect the efficiency with which N will be used. This study aimed to evaluate how the immediate and residual effect of 16 applications of animal manure reflects on yields of black oat and corn rotation system, as well as N use efficiency. The study was carried out in no-tillage from 2004 to 2016 in Brazil. The treatments were pig slurry (PS), dairy slurry (DS), pig deep-litter (PL), mineral fertilizer (MF), and control (C). Prior to the sowing of black oat, in which 16 animal manure applications had already been made, an area of the soil was delimited where the treatments were not applied. This area was referred to as unfertilized (U) soil. Applications were carried out in the remaining area and were referred to as fertilized (F) soil. The highest dry matter yield and N accumulation in black oat and corn were found in F soils treated with DS and PL, respectively. In corn, the highest grain yield and N accumulation in grains were found with DS and PS. In U soil, the 16 applications (of DS especially) resulted in yields and N accumulation greater than the control and MF, but lower than those in F. The highest N use efficiency was found with DS. The history of animal manure applications was not enough to rule out additional applications in the following years.

RESUMO OBJETIVO: Embora o relaxamento seja recomendado como terapia complementar no tratamento da hipertensão, seus efeitos na modulação autonômica cardiovascular ainda não estão claros. O objetivo desse trabalho foi investigar os efeitos do relaxamento na modulação autonômica cardíaca de indivíduos hipertensos. MÉTODOS: Este estudo aleatorizado e controlado foi realizado no Laboratório de Hemodinâmica da Escola de Educação Física e Esporte da Universidade de São Paulo/Brasil. Dezesseis hipertensos (6-mulheres) e 14 normotensos controle (6-mulheres) não-obesos participaram de 2 sessões aleatórias: relaxamento (R) e controle (C). Os participantes permaneceram supinos por 55 minutos após as intervenções. Eletrocardiograma, pressão arterial batimento a batimento, e respiração foram adquiridos durante e após as intervenções para a posterior análise espectral autorregressiva das variabilidades do intervalo R-R e da pressão arterial. RESULTADOS: Hipertensos e normotensos apresentaram respostas semelhantes de modulação autonômica cardíaca durante e após as sessões experimentais. Durante o relaxamento, a baixa frequência e o equilíbrio simpatovagal da variabilidade da frequência cardíaca estavam significantemente mais baixas no R do que no repouso supino em C. Quinze minutos após a intervenção a baixa frequência e o equilíbrio simpatovagal ainda estavam inferiores em R do que em C, o que ocorreu também nos 35 minutos, mas não aos 55 minutos. A variabilidade da pressão arterial foi semelhante entre as sessões. CONCLUSÃO: O relaxamento savasana reduz a modulação autonômica simpática cardíaca de hipertensos; essa redução perdura por pelo menos 35 minutos após o término do relaxamento e não é prejudicada nos hipertensos quando comparados com normotensos. Dessa forma, o relaxamento savasana tem efeitos positivos na modulação autonômica cardíaca de hipertensos e pode ser incluído como estratégia para o tratamento não-medicamentoso da hipertensão arterial.

OBJECTIVE: Although relaxation is recommended as complementary therapy for hypertension, its post-intervention cardiovascular autonomic effects are unclear. The objective of this research was to investigate the effects of savasana relaxation on cardiovascular autonomic modulation in hypertensive patients. METHODS: This randomized controlled trial was performed at the Hemodynamic Laboratory of the Physical Education School of the University of São Paulo/Brazil. Sixteen hypertensive (6-women) and 14 normotensive patients (6-women) non-obese subjects participated in 2 random sessions: savasana relaxation and control. Patients remained supine for 55 min after interventions. Electrocardiogram, beat-to-beat blood pressure and respiration were acquired during and after interventions for posterior autoregressive spectral analysis of the R-R interval and blood pressure variability. RESULTS: Hypertensive and normotensive patients presented similar cardiac autonomic modulation responses during and after experimental sessions. During relaxation, low frequency and sympathovagal balance were significantly lower in the Relaxation sessions than during supine rest in the Control sessions. Fifteen minutes after interventions, low frequency and sympathovagal balance were still lower in Relaxation than in Control, and remained lower for 35 min; at 55 min, the variables were similar between sessions. Systolic blood pressure variability did not differ among sessions. CONCLUSIONS: Savasana Relaxation decreases cardiac sympathetic autonomic modulation after its performance in hypertensive patients; this reduction lasts at least 35 minutes and is not blunted in hypertensive patients when compared to normotensive controls. Thus, savasana relaxation has positive effects on cardiac autonomic modulation of hypertensive patients, and may be included as a strategy for the non-drug treatment of hypertension.

O objetivo foi identificar subtipos do Vírus da Imunodeficiência Humana tipo-1 (HIV-1) e analisar a presença de mutações/polimorfismos nas regiões da protease (PR) e transcriptase reversa (TR) de 48 pacientes virgens de tratamento atendidos no município de Maringá, Paraná, Brasil. O sequenciamento foi conduzido usando produtos de nested PCR dos genes da PR, TR parcial e group-specific antigen gene (gag) de RNA retrotranscrito. A interpretação da resistência transmitida foi realizada segundo o algoritmo Surveillance Drug Resistance Mutation List (SDRM). As análises filogenética e SimPlot dos segmentos concatenados classificaram as sequências como subtipo B 19/48 (39,6%), subtipo C 12/48 (25%), subtipo F 4/48 (8,3%), com 13/48 (27,1%) formas recombinantes. A maioria das formas recombinantes era mosaicos B (B/F 12,5%, B/C 10,4%), com um C/F (2,1%) e um mosaico complexo B/C/F (2,1%). A prevalência de resistência transmitida foi de 4,2% (2,1% para ITRN e 2,1% para IP). Baixos níveis de resistência transmitida foram encontrados nesse estudo, 2/48 (2,1% para INTR e 2,1% para IP). Esses achados, embora preliminares, podem contribuir no monitoramento da epidemia de HIV na região.

The objective of this study is to identify subtypes of Human Immunodeficiency Virus type 1 (HIV-1) and to analyze the presence of mutations associated to antiretroviral resistance in the protease (PR) and reverse transcriptase (RT) regions from 48 HIV-1 positive treatment naïve patients from an outpatient clinic in Maringá, Paraná, Brazil. Sequencing was conducted using PR, partial RT and group-specific antigen gene (gag) nested PCR products from retrotranscribed RNA. Transmitted resistance was determined according to the Surveillance Drug Resistance Mutation List (SDRM) algorithm. Phylogenetic and SimPlot analysis of concatenated genetic segments classified sequences as subtype B 19/48 (39.6%), subtype C 12/48 (25%), subtype F 4/48 (8.3%), with 13/48 (27.1%) recombinant forms. Most recombinant forms were B mosaics (B/F 12.5%, B/C 10.4%), with one C/F (2.1%) and one complex B/C/F mosaic (2.1%). Low levels of transmitted resistance were found in this study, 2/48 (2.1% to NRTIs and 2.1% for PI). This preliminary data may subsidize the monitoring of the HIV evolution in the region.

CONTEXTUALIZAÇÃO: Exercícios para fortalecimento do glúteo máximo são empregados na prática clínica para tratamento de disfunções da coluna lombar e sacroilíaca. Entretanto, não foram encontrados estudos que investigaram os melhores exercícios para ativar esse músculo de forma a justificar a sua utilização na prática fisioterapêutica. OBJETIVO: Quantificar a atividade eletromiográgica (EMG) dos músculos glúteo máximo e semitendinoso durante quatro modalidades de exercícios terapêuticos. MÉTODOS: Trinta e um participantes (16 homens, 15 mulheres) foram selecionados. A atividade EMG do glúteo máximo e semitendinoso foi registrada e quantificada enquanto os indivíduos realizaram quatro modalidades de exercícios terapêuticos envolvendo extensão ativa do quadril em prono em quatro posições: joelho em extensão, joelho em flexão, rotação lateral do quadril e extensão do joelho e rotação lateral do quadril e flexão do joelho. RESULTADOS: ANOVA medidas repetidas revelou que a adição de flexão do joelho ou rotação lateral do quadril aumentou a atividade do glúteo máximo, enquanto os exercícios realizados com extensão do joelho resultaram na redução da sua ativação. Por outro lado, os exercícios realizados com extensão do joelho aumentaram a atividade do semitendinoso. CONCLUSÕES: Os exercícios realizados com flexão de joelho e/ou rotação lateral do quadril demonstraram ser opções efetivas para ativação do glúteo máximo, mas resultaram em redução da atividade do semitendinoso. Os exercícios realizados com extensão do joelho demonstraram ser uma boa opção para ativar o semitendinoso.

BACKGROUND: Gluteus maximus strengthening exercises are employed in clinical practice as options for the treatment of low back and sacroiliac disorders. However, no studies were found that investigated which were the best exercises to activate this muscle and justify its employment in physical therapy practice. OBJECTIVE: To quantify the electromyographic (EMG) activity of the gluteus maximus and semitendinosus muscles during four modalities of therapeutic exercises. METHODS: Thirty-one participants (16 men, 15 women) were selected. The EMG activities of the gluteus maximus and semitendinosus was recorded and quantified while the participants performed four modalities of therapeutic exercises, involving active prone hip extension in four positions: knee extension (KE), knee flexion (KF), lateral hip rotation and knee extension (LHRKE), and lateral hip rotation and knee flexion (LHR-KF). RESULTS: Repeated-measures ANOVAs showed that the addition of KF or LHR increased gluteus maximus activity, whereas, KE decreased the activation of this muscle. In contrast, the exercises performed with KE increased semitendinosus activity. CONCLUSIONS: Exercises performed with KF or LHR, or a combination of the two, may be effective choices for gluteus maximus strengthening, however both KF and LHR decreased semitendinosus activity. The exercises performed with KE appeared to be an acceptable choice for semitendinosus activation.

As in many areas of Brazil, the AIDS epidemic in Curitiba is relatively stable, but surveillance is important to support public policy. The molecular characteristics of HIV may be instrumental for monitoring epidemic trends. We evaluated plasma HIV-1 RNA (n = 37) from 38 cases presenting with positive serology, who were among 820 consenting volunteers visiting the downtown counselling and serology testing centre. Seroprevalence was 4.6% (CI 95% 3.2-6.3) and the estimated HIV incidence, as defined by the BED assay, was 2.86 persons/years (CI 95% 1.04-4.68). An additional set of contemporaneous, anonymous samples from a local laboratory was also analysed (n = 20). Regions of the HIV-1 polymerase (n = 57) and envelope (n = 34) were evaluated for subtyping, determination of mosaic structure, primary drug resistance mutations (pDRM), envelope V3 loop motifs and amino acid signatures related to viral tropism. HIV-1 clade B was observed in 53% of cases; HIV-1C in 30% and BC mosaics in 14%, with one F genome and one CF mosaic. Clade C infection was associated with recent infections among males (p < 0.03). Stanford surveillance pDRM was observed in 8.8% of sequences, with 7% showing high level resistance to at least one antiretroviral drug. Tropism for CXCR4 co-receptor was predicted in 18% of envelope sequences, which were exclusively among clade B genomes and cases with serological reactivity to chronic infection.