Arbovirus investigation in patients from Mato Grosso during Zika and Chikungunya virus introdution in Brazil, 2015-2016.

Arboviruses have become a major public health concern in Brazil, especially after Zika virus (ZIKV) and Chikungunya virus (CHIKV) introduction, leading to massive epidemics. We conducted an investigation of arboviruses in patients with acute febrile illness for less than five days in Mato Grosso state (MT) during the period of ZIKV and CHIKV dissemination in Brazil. To achieve that, 453 human serum samples of patients suspected of Dengue (DENV), Yellow Fever (YFV), ZIKV or CHIKV collected in health units of 31 cities of MT were subjected to RT-PCR protocols for 10 flaviviruses, 5 alphaviruses and orthobunyaviruses from Simbu serogroup, nucleotide sequencing and viral isolation. Regarding flaviviruses, five (1.1%) patients were infected with DENV-1 genotype V, 22 (4.4%) with DENV-4 genotype II, 3 (0.7%) with YFV South American genotype II and five (1.1%) with ZIKV Asian genotype. The first human case of ZIKV in MT was detected in this study during August, 2015 in Tapurah. Alphaviruses were detected in 2 (0.4%) patients infected with CHIKV genotype ECSA, 1 (0.2%) with Madariaga (EEEV) lineage III and 34 (7.5%) with Mayaro (MAYV) genotype L. Four (11.4%) patients presented dual infections with DENV-1/ZIKV, DENV-1/DENV4, DENV-4/MAYV and ZIKV/MAYV. The majority - 13/34 positive for MAYV, one for Madariaga virus - are residents in Várzea Grande (VG), metropolitan region of Cuiabá, capital of MT. The first CHIKV infection in MT was detected in this study in Mirassol D'Oeste, during July, 2015. In addition, 20 (4.4%) patients were positive for OROV Segment S genotype IA. These results reinforce the variation in arboviruses frequency and distribution during outbreaks, highlinghing the importance of differential diagnosis to identify agents silently co-circulating with major health problem arboviruses.

Diallel and prediction (REML/BLUP) for yield components in intervarietal maize hybrids.

Genetic improvement is essential to achieve increments in maize (Zea mays L.) grain yield components. It may be obtained through crosses, which enable to exploit the effects of intervarietal heterosis, allelic complementarity, as well as gene actions and effects. This study estimated the components of variance and genetic parameters (REML/BLUP) of an intervarietal diallel to select and predict the best genotypes for maize yield components. The experimental design was randomized blocks containing 60 intervarietal maize hybrids arranged in three repetitions. They were obtained through intervarietal crosses and evaluated in a diallel scheme, where 14 open-pollinated varieties were designated as parentals. Thus, 10 crosses were performed for each hybrid combination to obtain the number of seeds necessary for the competition test. The measured traits were: grain volume relative index, the mass of one hundred grains, and grain yield. The male parents and the additive genetic fraction were determinants for grain volume relative index. Mass of one hundred grains and grain yield were defined by the specific combining ability, and female parents revealed low narrow sense heritability. The female parent Taquarão and male parent Argentino Amarelo presented the best general combining abilities for the measured traits. The specific combining abilities were expressed for crosses AL 25 x Dente de Ouro Roxo, AL 25 x BRS Pampeano, and Taquarão x Argentino Branco. Genetic estimates and predictions were consistent and applicable to breeding programs and could be applied in future quantitative genetic studies of maize.

Estimates of genetic parameters and genotypic values prediction in maize landrace populations by REML/BLUP procedure.

The REML/BLUP statistics are analyses that can be used as selective criteria in the routine of maize breeding programs. The present study aims to determine the genetic potential in crosses of landrace populations applying the REML/BLUP methodology, and to identify populations for the synthesis of new populations and intrapopulation selection for family farming systems, as well as genetic constitutions for use in maize breeding programs. Nine top cross hybrids obtained in the 2012/2013 harvest were evaluated along with their testator, the landraces used as parents, and four commercial hybrids, in a randomized block design, with information taken from the average of each plot. The evaluated traits were: leaf angle, number of ramifications of the tassel, spike insertion height, plant height, spike diameter, number of grains per spike, mass of grains per spike, spike mass, spike length, prolificity, mass of one hundred grains, and grain yield per plot. The data were analyzed using the Selegen-REML/BLUP software. The top cross hybrids Cateto Branco x Planalto, Amarelão x Planalto and the population Cateto Branco are ranked among the ten best crosses, simultaneously, for the traits: leaf angle, number of ramifications of the tassel, spike insertion height, and plant height (Cateto Branco x Planalto), and leaf angle, spike insertion height, and plant height (Amarelão x Planalto and Cateto Branco). The top cross hybrids Criolão x Planalto, Branco 8 Carreiras x Planalto, Caiano Rajado x Planalto, Amarelão x Planalto, Branco Roxo Índio x Planalto stand out for their high genotypic value of the individual BLUP mean components among the ten best genotypes for grain yield, and by combining three or more traits of interest together, being, for effects of selection, the most indicated.

Contribution of morphoagronomic traits to grain yield and earliness in grain sorghum.

Given the importance of selecting lines to obtain hybrids, we aimed to verify the relationship between morphological traits that can be used as the criteria for the selection of sorghum lines with high grain yield and earliness. A total of 18 traits were evaluated in 160 sorghum elite lines, in an incomplete block design with two replicates. A correlation network was used to graphically express the estimates of phenotypic and genotypic correlations between the traits. Two path analyses were processed, the first considering grain yield and the second considering flowering as the principle dependent variable. In general, most of the variation in the grain yield and flowering of sorghum lines was explained by the traits evaluated. Selecting sorghum lines with greater width of the third leaf blade from flag leaf, panicle weight, and panicle harvest index might lead to increased grain yield, and selecting sorghum genotypes with higher plant height might lead to reduced earliness and increased grain yield. Thus, the results suggest the establishment of selection indices aiming at simultaneously increasing the grain yield and earliness in sorghum genotypes.

Genetic progress in homogeneous regions of wheat cultivation in Rio Grande do Sul State, Brazil.

The State of Rio Grande do Sul (RS) stands out as the largest wheat producer in Brazil. Wheat is the most emphasized winter cereal in RS, attracting public and private investments directed to wheat genetic breeding. The study of genetic progress should be performed routinely at breeding programs to study the behavior of cultivars developed for homogeneous regions of cultivation. The objectives of this study were: 1) to evaluate the genetic progress of wheat grain yield in RS; 2) to evaluate the influence of cultivar competition trial stratification in homogeneous regions of cultivation on the study of genetic progress. Grain yield data of 122 wheat cultivars evaluated in 137 trials arranged in randomized block design with three or four replications were used. Field trials were carried out in 23 locations in RS divided into two homogeneous regions during the period from 2002 to 2013. Genetic progress for RS and homogeneous regions was studied utilizing the method proposed by Vencovsky. Annual genetic progress for wheat grain yield during the period of 12 years in the State of RS was 2.86%, oscillating between homogeneous regions of cultivation. The difference of annual genetic progress in region 1 (1.82%) in relation to region 2 (4.38%) justifies the study of genetic progress by homogeneous regions of cultivation.

REML/BLUP and sequential path analysis in estimating genotypic values and interrelationships among simple maize grain yield-related traits.

Methodologies using restricted maximum likelihood/best linear unbiased prediction (REML/BLUP) in combination with sequential path analysis in maize are still limited in the literature. Therefore, the aims of this study were: i) to use REML/BLUP-based procedures in order to estimate variance components, genetic parameters, and genotypic values of simple maize hybrids, and ii) to fit stepwise regressions considering genotypic values to form a path diagram with multi-order predictors and minimum multicollinearity that explains the relationships of cause and effect among grain yield-related traits. Fifteen commercial simple maize hybrids were evaluated in multi-environment trials in a randomized complete block design with four replications. The environmental variance (78.80%) and genotype-vs-environment variance (20.83%) accounted for more than 99% of the phenotypic variance of grain yield, which difficult the direct selection of breeders for this trait. The sequential path analysis model allowed the selection of traits with high explanatory power and minimum multicollinearity, resulting in models with elevated fit (R2 > 0.9 and ε < 0.3). The number of kernels per ear (NKE) and thousand-kernel weight (TKW) are the traits with the largest direct effects on grain yield (r = 0.66 and 0.73, respectively). The high accuracy of selection (0.86 and 0.89) associated with the high heritability of the average (0.732 and 0.794) for NKE and TKW, respectively, indicated good reliability and prospects of success in the indirect selection of hybrids with high-yield potential through these traits. The negative direct effect of NKE on TKW (r = -0.856), however, must be considered. The joint use of mixed models and sequential path analysis is effective in the evaluation of maize-breeding trials.

Chronic myelomonocytic leukemia mimicking hematologic systemic lupus erythematosus.

The relationship between primary hematologic disease and rheumatologic manifestations is well known, especially acute lymphocytic leukemia, lymphomas, plasma cell dyscrasias and myelodysplastic syndrome (MDS). Currently, more has been described about autoimmune manifestations in chronic myelomonocytic leukemia (CMML). Many different clinical scenarios may lead a patient with MDS/CMML initially to seek a rheumatological unit. Autoimmune features such as polymyalgia rheumatic symptoms, myositis, neutrophilic dermatosis, cutaneous vasculitis and positive antinuclear antibodies (ANA) are some examples of clinical presentation of MDS/CMML. Moreover, peripheral cytopenias are a common initial presentation both for systemic lupus erythematous (SLE) and MDS/CMML. The aim of this study was to describe a case of an elderly woman with thrombocytopenia and positivity of antibodies to anti-extractable nuclear antigens (anti-ENA) as initial manifestation of CMML mimicking SLE, and to present some clues that encourage the clinician to perform a bone marrow study in such a clinical scenario.

Genetic progress in oat associated with fungicide use in Rio Grande do Sul, Brazil.

The State of Rio Grande do Sul (RS) is the largest producer of oat in Brazil with the aid of consolidated breeding programs, which are constantly releasing new cultivars. The main objectives of this study were to: 1) evaluate the annual genetic progress in grain yield and hectoliter weight of the oat cultivars in RS, with and without fungicide use on aerial parts of plants; and 2) evaluate the efficiency of oat breeding programs in introducing disease-resistant genes in the released cultivars through network yield trials conducted with and without fungicide use on aerial plant parts. The data on grain yield and hectoliter weight were obtained from 89 competition field trials of oat cultivars carried out from 2007 to 2014 in nine municipalities of RS. Of the total 89 trials, 44 were carried out with fungicide application on aerial plant parts and 45 were carried out without fungicide application. The annual genetic progress in oat cultivars was studied using the methodology proposed by Vencovsky (1988). The annual genetic progress in oat grain yield was 1.02% with fungicide use and 4.02% without fungicide use during the eight-year study period in RS. The annual genetic progress with respect to the hectoliter weight was 0.08% for trials with fungicide use and 0.71% for trials without fungicide use. Performing network yield trials with and without fungicide use on the aerial plants parts is a feasible method to evaluate the efficiency of oat breeding programs in introducing disease-resistant genes in the released cultivars.

Solitary lung cavities: CT findings in malignant and non-malignant disease.

AIM: To assess the computed tomography (CT) findings of solitary cavitary lesions which could potentially aid in differentiating malignant from non-malignant lung disease. MATERIALS AND METHODS: A retrospective study of patients diagnosed with a solitary lung cavity at two university hospitals, who underwent multidetector CT examinations of the chest between 2012 and 2014, was performed. Lesions were evaluated for maximum diameter, maximum wall thickness, and associated findings. Statistical analyses were then conducted and a receiver operating characteristic (ROC) curve was calculated to select the most accurate cut-off value for malignant and non-malignant lesions. RESULTS: CT and clinical records from 96 patients were reviewed. The most frequent aetiologies of non-malignant and malignant lung cavities were mycobacterial infection sequelae (50%, 33/66) and primary lung carcinoma (94%, 28/30), respectively. Significant differences (p<0.05) were found between malignant and non-malignant cases when comparing the averages of maximum wall thickness (15.2 and 7.8 mm, respectively) and maximum diameter of lesions (51 and 35 mm, respectively). The presence of either perilesional consolidation or centrilobular nodules favoured the diagnosis of non-malignant conditions (p<0.05). Maximum wall thicknesses thresholds of ≤7 or ≥24 mm were the most accurate in suggesting non-malignant and malignant aetiologies, respectively. CONCLUSION: Malignant and non-malignant solitary lung cavities differ significantly at CT. Non-malignant lesions tend to exhibit thinner walls, but more perilesional consolidation and centrilobular nodules than malignant lesions. The results reveal that maximum wall thicknesses of ≤7 and ≥24 mm are indicative of non-malignant and malignant disease, respectively.

Pre-emptive pediatric kidney transplantation or not?

BACKGROUND: Kidney transplantation prior to dialysis (pre-emptive kidney transplantation, PKT) has been controversial because of the paucity of clinical evidence to clarify the risks and benefits of PKT. Several authors have confirmed a significant advantage of PKT in the treatment of patients with end-stage renal disease (ESRD). The aim of this study was to examine the characteristics of patients who received PKT or non-pre-emptive kidney transplant (NPKT). METHODS: We used a cohort of 323 consecutive kidney-transplanted children (53% boys) from Hospital da Criança Santo Antonio, Porto Alegre, Brazil, who underwent transplantation between January 2000 and December 2010. RESULTS: The main causes of ESRD were congenital anomalies of the kidney and urinary tract (CAKUT) (39%) and glomerulopathies (27.5%). The 12-, 36-, 60-, and 90-months death-censored graft survival rates were 97%, 92%, 86%, and 76%, respectively, in the PKT group, and 87%, 79%, 72%, and 65% in the NPKT group (P < .05). CONCLUSIONS: The results of this study suggest that pre-emptive transplantation is beneficial (hazard ratio = 0.37; 95% confidence interval: 0.18-0.82). The main causes of graft loss (n = 67) were recurrence of primary disease (21%), chronic allograft injury (17%), and death with a functioning graft (16%). We recommend PKT as a better choice for transplantation whenever possible to minimize ESRD morbidity and provide better long-term patient and graft survival.

Collaborative Brazilian Pediatric Renal Transplant Registry (CoBrazPed-RTx): A Report From 2004 to 2013.

BACKGROUND: The Collaborative Brazilian Pediatric Renal Transplant Registry started in 2004 as a multicenter initiative aiming to analyze, report, and share the results of pediatric kidney transplantation in Brazil. Data from all pediatric kidney transplants performed between January 2004 and December 2013 were recorded electronically and periodically updated. All patients under 18 years old from the participating centers were enrolled. Demographic data, etiology of chronic kidney disease, and patient and graft survival were analyzed. From a total of 2443 pediatric kidney transplants performed in Brazil during the study period, we report data from 1751 pediatric renal transplants performed in 13 centers enrolled in the collaborative study. Median age at transplantation was 12.4 years, and most of recipients were male (56%). The most common underlying renal etiologies were obstructive uropathy (31%) and glomerulopathy (26%). METHODS: According to donor source, 1155 (66%) of transplants were performed with deceased donors (DD). Initial immunosuppression consisted mainly of tacrolimus, mycophenolate, steroids, and induction therapy with anti-IL-2R antibodies. RESULTS: One-year graft survival (death-censored) was 93% and 90% (log rank test, P < .01), respectively, for living donor (LD) and DD. Graft losses (15%) were most frequently caused by vascular thrombosis, chronic allograft nephropathy, death with functioning kidney, acute rejection, and recurrent renal disease. Recipients of DD had 2.02 (95% confidence interval: 1.14-3.59) times the hazard of graft loss compared with those of LD (P = .015). Patient survival rates at 1 and 5 years were 98% and 97% for LD and 97% and 93% for DD, respectively. The mortality rate was 3.8%, mainly as the result of infection and cardiovascular disease. CONCLUSIONS: The results of this collaborative pediatric transplant study are comparable to international registries. Our effort has been able to maintain an exchange of information, both among the participating centers and with other international registries.

TH17 cells, interleukin-17 and interferon-γ in patients and households contacts of leprosy with multibacillary and paucibacillary forms before and after the start of chemotherapy treatment.

BACKGROUND: Studies on the immunology of leprosy are fundamental to the understanding of the various forms of clinical manifestation of the disease. In some diseases, lymphocytes TH17 and one of its key cytokines, interleukin-17 has been shown to be essential in developing an effective immune response. In leprosy, involvement of lymphocyte TH17 and interleukin-17 remains understudied. OBJECTIVES: This study is the first investigation to examine the association between TH17 cells, interleukin-17 and interferon- γ in patients and households contacts of leprosy. METHODS: To document the participation of TH17 cells and interleukin-17 in the immunology of leprosy, to observe the behavior of interferon-γ in relation to interleukin-17 and to verify the differences found between individuals paucibacillary, multibacillary and household contacts, we analyzed samples peripheral blood to identify TH-17 cells, interleukin-17 and IFN-γ; establishing relationships between all the groups. RESULTS: There was a significant difference in the results found in the comparison between the paucibacillary and multibacillary groups of patients (P < 0.001), as well with the household contacts (P < 0.005). The polychemotherapeutic treatment modified the profile of immune response in multibacillary patients compared to what was observed before the start of treatment. CONCLUSION: The principal finding was that TH17 lymphocytes and interleukin-17 actively participating in the immune response of Hansen's disease as well these cells can stimulate the cellular immunity.

Pseudotumoral form of cerebral Schistosomiasis Mansoni.

The authors report a case of 36-year-old woman presented with epileptic seizures and headaches. Magnetic resonance imaging (MRI) revealed an enhancing lesion with surrounding edema and mild mass effect in the left frontal lobe. Stereotactic brain biopsy demonstrated intraparenchymal granulomas surrounding S. mansoni eggs. Praziquantel was started (60mg/kg of body weight, in a single dose), followed by Prednisone (80mg/day) for seven days to treat the cerebral edema. The patient's symptoms resolved following medical treatment and the follow-up MRI yielded normal findings.

Molecular characterization of cajá, Spondias mombin (Anacardiaceae), by RAPD markers.

The arboreal species Spondias mombin L. (Anacardiaceae) is widely distributed in Brazil, where the fruits, known by the common name of cajá, are an important commercial commodity. We evaluated genetic variability among 32 cajá accessions of the Germplasm Collection of Embrapa Meio-Norte using RAPD technique. Reaction conditions for efficient RAPD amplifications were optimized in preliminary tests, and primers were selected from a set designed by the University of British Columbia on the basis of high levels of polymorphism and adequate band resolution. The 21 primers employed in the final analysis produced 145 fragments, 79% of which were polymorphic. Based on the RAPD data, a dendrogram was constructed using the unweighted pair group method with arithmetic mean clustering technique. The 32 cajá accessions were classified into three main groups with a mean genetic similarity of 68.8%. Group I comprised 26 accessions (74.1% similarity), and group II included five accessions (74.0% similarity), while group III consisted of one accession (BGC 06), which exhibited the lowest similarity coefficients. Accessions BGC 06 and BGC 31 were the most unrelated and, hence, most suitable for initial crossings in order to obtain high levels of segregation. We concluded, based on the repeatability and reproducibility tests, that the RAPD technique is reliable and efficient for revealing the genetic diversity of cajá accessions, which will be useful for genetic improvement programs.

RAPD analysis of the genetic diversity of mango (Mangifera indica) germplasm in Brazil.

We evaluated genetic variability of mango (Mangifera indica) accessions maintained in the Active Germplasm Bank of Embrapa Meio-Norte in Teresina, Piauí, Brazil, using RAPDs. Among these accessions, 35 originated from plantings in Brazil, six from the USA and one from India. Genomic DNA, extracted from leaf material using a commercial purification kit, was subjected to PCR with the primers A01, A09, G03, G10, N05, and M16. Fifty-five polymorphic loci were identified, with mean of 9.16 ± 3.31 bands per primer and 100% polymorphism. Application of unweighted pair group method using arithmetic average cluster analysis demonstrated five genotypic groups among the accessions examined. The genotypes Rosa 41, Rosa 48 and Rosa 49 were highly similar (94% similarity), whereas genotypes Sensation and Rosa 18 were the most divergent (only 7% similarity). The mango accessions were found to have considerable genetic variability, demonstrating the importance of analyzing each genotype in a collection in order to efficiently maintain the germplasm collection.

IFNG +874 T>A single nucleotide polymorphism is associated with leprosy among Brazilians.

Leprosy is a chronic infectious disease caused by Mycobacterium leprae, a low virulence mycobacterium, and the outcome of disease is dependent on the host genetics for either susceptibility per se or severity. The IFNG gene codes for interferon-γ (IFN-γ), a cytokine that plays a key role in host defense against intracellular pathogens. Indeed, single nucleotide polymorphisms (SNPs) in IFNG have been evaluated in several genetic epidemiological studies, and the SNP +874T>A, the +874T allele, more specifically, has been associated with protection against infectious diseases, especially tuberculosis. Here, we evaluated the association of the IFNG locus with leprosy enrolling 2,125 Brazilian subjects. First, we conducted a case-control study with subjects recruited from the state of São Paulo, using the +874 T>A (rs2430561), +2109 A>G (rs1861494) and rs2069727 SNPs. Then, a second study including 1,370 individuals from Rio de Janeiro was conducted. Results of the case-control studies have shown a protective effect for +874T carriers (OR(adjusted) = 0.75; p = 0.005 for both studies combined), which was corroborated when these studies were compared with literature data. No association was found between the SNP +874T>A and the quantitative Mitsuda response. Nevertheless, the spontaneous IFN-γ release by peripheral blood mononuclear cells was higher among +874T carriers. The results shown here along with a previously reported meta-analysis of tuberculosis studies indicate that the SNP +874T>A plays a role in resistance to mycobacterial diseases.

A simple approach to assess VT during a field walk test.

We aimed to examine whether the ventilatory threshold (VT) during an incremental shuttle walk test (ISWT) could be determined using heart rate variability (HRV) analysis. Further aims were to assess variables capable of predicting performance in the ISWT and the intensity of this test. Beat-to-beat RR intervals and gas exchange values in 10 healthy subjects (31-83 years; 7 men) were collected during the ISWT. The ventilatory equivalent was used to assess VT from respiratory components. To determine the HRV threshold (HRVT), the instantaneous beat-to-beat variability values of the RR intervals at each stage of exercise were graphically plotted against walking speed (WS). The oxygen consumption at HRVT was calculated (VO2HRVT). No significant differences were found between walking speed (WS) at VT and WS at HRVT (5.04±1.00 vs. 5.10±1.04 km/h; p=0.89). Linear regression analysis revealed a strong correlation between VO2VT and VO2HRVT (r(2)=0.896). The Bland and Altman plot analysis revealed an agreement between VO2VT and VO2HRVT (-0.05; 95%CI: -0.30-0.20 L/min). Thus, the VT can be assessed during the ISWT using a simple heart monitor. The ISWT may be a useful tool to assess exercise capacity and prescribe walking programs.

Effects of strain and age on ear wound healing and regeneration in mice

Round holes in the ears of MRL mice tend to close with characteristics of regeneration believed to be absent in other mouse strains (e.g., C57BL/6). We evaluated the kinetics and the histopathology of ear wound closure in young (8 weeks old) C57BL/6 and BALB/c mice. We also used middle-aged (40 weeks old) C57BL/6 mice to evaluate the influence of aging on this process. A circular through-and-through hole was made in the ear, photographs were taken at different times after injury and wound area was measured with digital analysis software. The percentages of closed area measured on day 100 were: 23.57 ± 8.66 percent for young BALB/c mice, 56.47 ± 7.39 percent for young C57BL/6 mice, and 75.31 ± 23.65 percent for middle-aged C57BL/6 mice. Mice were sacrificed on days 1, 3, 5, 25, 44, and 100 for histological evaluation with hematoxylin and eosin, Gomori’s trichrome, periodic acid-Schiff, or picrosirius red staining. In young mice of both strains, healing included re-epithelialization, chondrogenesis, myogenesis, and collagen deposition. Young C57BL/6 and BALB/c mice differed in the organization of collagen fibers visualized using picrosirius-polarization. Sebaceous glands and hair follicles regenerated and chondrogenesis was greater in young C57BL/6 mice. In middle-aged C57BL/6 mice all aspects of regeneration were depressed. The characteristics of regeneration were present during ear wound healing in both young BALB/c and young C57BL/6 mice although they differed in intensity and pattern. Greater ear wound closure in middle-aged C57BL/6 mice was not correlated with regeneration.

Effects of strain and age on ear wound healing and regeneration in mice.

Round holes in the ears of MRL mice tend to close with characteristics of regeneration believed to be absent in other mouse strains (e.g., C57BL/6). We evaluated the kinetics and the histopathology of ear wound closure in young (8 weeks old) C57BL/6 and BALB/c mice. We also used middle-aged (40 weeks old) C57BL/6 mice to evaluate the influence of aging on this process. A circular through-and-through hole was made in the ear, photographs were taken at different times after injury and wound area was measured with digital analysis software. The percentages of closed area measured on day 100 were: 23.57 +/- 8.66% for young BALB/c mice, 56.47 +/- 7.39% for young C57BL/6 mice, and 75.31 +/- 23.65% for middle-aged C57BL/6 mice. Mice were sacrificed on days 1, 3, 5, 25, 44, and 100 for histological evaluation with hematoxylin and eosin, Gomori's trichrome, periodic acid-Schiff, or picrosirius red staining. In young mice of both strains, healing included re-epithelialization, chondrogenesis, myogenesis, and collagen deposition. Young C57BL/6 and BALB/c mice differed in the organization of collagen fibers visualized using picrosirius-polarization. Sebaceous glands and hair follicles regenerated and chondrogenesis was greater in young C57BL/6 mice. In middle-aged C57BL/6 mice all aspects of regeneration were depressed. The characteristics of regeneration were present during ear wound healing in both young BALB/c and young C57BL/6 mice although they differed in intensity and pattern. Greater ear wound closure in middle-aged C57BL/6 mice was not correlated with regeneration.