Puberty attainment and reproductive performance of yearling Bos indicus-influenced heifers after two sequential treatments with progesterone.

Number of pubertal heifers at time of breeding season initiation is a primary determinant to pregnancy success during the breeding season. It was hypothesized that pre-breeding progesterone (P4) supplementation (induction) would increase the number of heifers pubertal at the time of imposing estrous synchronization treatment regimens and P/AI. Yearling, Bos indicus-influenced (n = 577) or Bos indicus (n = 174) heifers were or were not treated with P4 (CIDR and Non-CIDR, respectively) for 10 d starting on D-23 (D0 = TAI). Presence of a CL on D-33 or D-23 was considered to indicate heifers were pubertal. On D-13, there was a PGF analogue administered. On D-9, there was treatment with GnRH analogue, 6d-CIDR and PGF. There were inseminations based on estrus (D-2 to D0) or TAI on D0 for non-estrous animals. There were 5.2 % and 62.9 % purebred and crossbred heifers pubertal, respectively. Proportion of prepubertal crossbred than purebred heifers with CL on D-3 was greater as a result of imposing the pubertal induction regimen (P < 0.05 and P> 0.10, respectively). Regardless of puberty status, proportion of heifers in estrus prior to AI in the CIDR group was similar to the heifers of the Non-CIDR group for crossbreds and purebreds. Similarly, P/AI of CIDR group was similar to the Non-CIDR group for crossbreds and purebreds. In summary, imposing the pubertal induction regimen hastened attainment of puberty in yearling crossbred, but not purebred heifers. Puberty induction did not affect estrous response, neither fertility after imposing an estrous synchronization treatment regimen.

Type I interferon receptors and interferon-τ-stimulated genes in peripheral blood mononuclear cells and polymorphonuclear leucocytes during early pregnancy in beef heifers.

This study characterised the expression of interferon (IFN)-τ-stimulated genes (ISGs) and Type I IFN receptors in circulating polymorphonuclear cells (PMNs) of beef heifers and compared it with expression in peripheral blood mononuclear cells (PBMCs) up to Day 20 of gestation. Nelore heifers (n=26) were subjected to fixed-time AI (FTAI) on Day 0. PMNs and PBMCs were isolated on Days 0, 10, 14, 16, 18 and 20 after FTAI. The abundance of target transcripts (ubiquitin-like protein (ISG15), 2'-5'-oligoadenylate synthetase 1 (OAS1), myxovirus resistance 1 (MX1), myxovirus resistance 2 (MX2), IFN receptor I (IFNAR1) and IFN receptor 2 (IFNAR2)) was determined using real-time quantitative polymerase chain reaction and compared between pregnant (n=8) and non-pregnant (n=9) females. In both PBMCs and PMNs, ISG15 and OAS1 expression was greater in pregnant than non-pregnant heifers on Days 18 and 20. There were no significant differences in the expression of ISGs between PBMCs and PMNs. A time effect on expression was found for IFNAR1 in PBMCs and IFNAR2 in PMNs, with decreased expression of both genes on Days 18 and 20. When the expression of these genes was compared between cell types only in pregnant heifers, IFNAR2 expression in PMNs had an earlier decrease when compared to its expression in PBMCs, starting from Day 18. In conclusion, PMNs do not respond earlier to the conceptus stimulus, and ISG15 and OAS1 expression in both PMNs and PBMCs can be used as a suitable marker for pregnancy diagnosis on Days 18 and 20. In addition, gestational status did not affect IFNAR1 and IFNAR2 expression, but IFNAR2 showed a distinct response between PMNs and PBMCs of pregnant heifers.

Identification of oral cavity abnormalities in pre-term and full-term newborns: a cross-sectional and comparative study.

PURPOSE: Compare maxillary labial frenum and lingual frenum topography, the ridges relationship and oral inclusion cysts occurrence between pre-term (PT) and full-term newborns (FT). METHODS: This cross-sectional and comparative study was conducted through the evaluation of 74 PT and 100 FT. Data were collected from medical records: mother age, gestational age, gender, height, weight, and delivery type. The variables were verified by Chi-square test and Mann-Whitney U test, at 5% significance level. RESULTS: Bohn's nodules and dental lamina cysts were more frequent in FT (P = 0.000). Epstein pearls occurrence was similar between FT and PT (P = 0.243). The lower alveolar ridge in distal position to the upper one was more prevalent in both groups. Abnormal upper labial frenum anatomy had been observed in 10.0% of FT. Upper labial frenum was attached in piriform papilla in 90.5% of PTG, whereas in FT, 61.0% were attached from crest of alveolar ridge to mucogingival line. CONCLUSIONS: PT oral cavity presented some peculiarities when compared with FT: maxillary labial frenum insertion in the Piriform papilla and palatal cysts more prevalent than alveolar cysts.

Maxillary arch dimensions in the first 6 months of life and their relationship with pacifier use.

PURPOSE: This study evaluated the maxillary arch dimensions at birth and 6 months of life, and to verify the influence of pacifier use on palatal development. METHODS: Maxillary arch impressions were taken from 80 full-term newborns, appropriate for gestational age, from 24 to 72 h of life, and repeated after 6 months. Anatomical references were mapped on dental casts and measurements were taken. Parents were questioned about feeding practices and sucking habits. Data were analysed by Kruskal-Wallis test and hierarchical mixed regression model (p < 0.05). RESULTS: Anterior maxillary segment (median [interquartile range]) presented an increase in transverse (Δ% = 81.90 [74.40-89.70]) and in length (Δ% = 76.20 [49.10-102.70]) relative growth. No differences were detected in palatine depth from birth (11.00 mm [10.30-11.70]) to 6 months (12.60 mm [12.0-13.90]). CONCLUSIONS: Pacifier use was positively related to the increase in anterior segment length. In the first 6 months of life, the greatest relative growth of the maxillary arch occurs in the anterior region, showing this growth does not occur homogeneously. The use of pacifiers came to influence this process, even before the eruption of the upper incisors.

In vitro evaluation of antagonism, modulation of cytokines and extracellular matrix proteins by Bifidobacterium strains.

A healthy skin provides a protective barrier against pathogenic micro-organisms. Recent studies have shown that probiotics, as those of Bifidobacterium genus, could act beneficially in dermatology, both when ingested and by topical use. In the present study, we evaluated by in vitro antagonism assays and using two skin cell lines the potential of four strains of Bifidobacterium spp. Among the four bifidobacteria, Bifidobacterium longum 51A was the only one able to inhibit the growth of the eight pathogenic indicators tested. Production of some cytokines and extracellular matrix proteins was determined when ccc or inactivated cells of the bifidobacteria were incubated with keratinocyte and/or fibroblast cell cultures. Significant results were observed only for IL-6, IL-8 and IL-18 production, and inactivated Bifidobacterium pseudolongum 1191A was the only one which significantly stimulated collagen production, whereas lumican was stimulated by treatments with live Bifidobacterium bifidum 1622A , B. longum 51A and B. pseudolongum 1191A . Highest adhesion and internalization capabilities were observed with B. bifidum 1622A and Bifidobacterium breve 1101A . Concluding, B. longum 51A was highlighted for its antagonistic capacity and B. bifidum 1622A and B. pseudolongum 1191A for stimulating the production of cytokines and proteins of the extracellular matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: The skin is the first line of defence against invasive micro-organisms, and its local microbiota provides additional protective functions based on antagonism against pathogenic micro-organisms and immunomodulation. Based on in vitro assays using Bifidobacterium spp. we demonstrated the antagonistic potential, as well as capacity in stimulating the production of cytokines and proteins of the extracellular matrix that these bacteria may exert on skin cells. This positive influence suggests the use of a consortium of these bifidobacteria in a topical product for dermatological treatments.

Optical coherence tomography applied to tests of skin care products in humans--a case study.

BACKGROUND/PURPOSE: When evaluating skin care products for human skin, quantitative test methods need to be simple, precise and reliable. Optical coherence tomography (OCT), provides high-resolution sectional images of translucent materials to a depth of a few millimeters, a technique usually applied to medical measurements in ophthalmology and dermatology. This study aimed to demonstrate the application of OCT as the main technique for monitoring changes in skin topography during tests of a wrinkle-reduction product in humans. METHODS: We used a commercial OCT apparatus to perform clinical examinations of skin roughness in treated and non-treated sites in the periorbital region of thirty human voluntaries who were using an anti-aging product commercially available: Natura Chronos® Flavonóides de Passiflora 45+ FPS15, from Natura Cosméticos, Brazil. Measurements were performed days 0, 7, 14 and 28 of treatment. Equipment and software allowed real-time recording of skin roughness parameters and wrinkle depths. RESULTS: The OCT measurements have allowed the monitoring of changes in skin roughness, which have shown reduction in treated sites around 10%. The obtained depth distributions also indicate reduction in the occurrence of wrinkles deeper than 170 µm. The verified results are consistent with those typically obtained after successful treatment with modern anti-aging products. CONCLUSION: By using the OCT technique, it was possible to quantify changes in skin roughness and in the distribution of depths of skin wrinkles, with adequate sensitivity. OCT imaging allows the direct visualization of the skin topography with resolution of micrometers, a reliable and interactive tool for clinical use. Therefore, for the first time, we demonstrated the use of OCT technique to verify the efficacy of cosmetic products in real time.

Two new Brazilian isolates of Bacillus thuringiensis toxic to Anticarsia gemmatalis (Lepidoptera: Noctuidae).

Bacillus thuringiensis is a bacterium used for biopesticides production and pest-resistant plants due to the synthesis of protein crystals by cry genes, which are effective in controlling several insect orders such as Lepidoptera. This work aimed at the evaluation and characterisation of two new B. thuringiensis isolates active against A. gemmatalis (Hübner 1818) larvae, which is the soybean major pest. The results showed that Bt117-4 isolate amplified fragments corresponding to cry2 and cry9 genes, and synthesised protein fragments equivalent to 130, 90 and 45 kDa. The Bt3146-4 isolate amplified DNA fragments corresponding to cry9 gene and synthesised protein fragments of 70, 58 and 38 kDa. Transmission electron microscopy revealed the presence of protein crystals in both isolates. CL50 with Cry purified proteins from Bt117-4 and Bt3146-4, corresponded to 0.195 and 0.191 µg larvae-1, respectively. The two B. thuringiensis isolates selected in this study were effective to control velvetbean caterpillar at laboratory conditions. Field tests should be carried on to develop new biopesticides formulation as well for cry genes resource for Anticarsia gemmatalis resistant transgenic plants.

Antinoceptive effect of triterpenoid alpha,beta-amyrin in rats on orofacial pain induced by formalin and capsaicin.

The effects of alpha,beta-amyrin, a pentacyclic triterpene isolated from Protium heptaphylum was investigated on rat model of orofacial pain induced by formalin or capsaicin. Rats were pretreated with alpha,beta-amyrin (10, 30, and 100mg/kg, i.p.), morphine (5mg/kg, s.c.) or vehicle (3% Tween 80), before formalin (20 microl, 1.5%) or capsaicin (20 microl, 1.5 microg) injection into the right vibrissa. In vehicle-treated controls, formalin induced a biphasic nociceptive face-rubbing behavioral response with an early first phase (0-5 min) and a late second phase (10-20 min) appearance, whereas capsaicin produced an immediate face-rubbing (grooming) behavior that was maximal at 10-20 min. Treatment with alpha,beta-amyrin or morphine significantly inhibited the face-rubbing response in both test models. While morphine produced significant antinociception in both phases of formalin test, alpha,beta-amyrin inhibited only the second phase response, more prominently at 30 mg/kg, in a naloxone-sensitive manner. In contrast, alpha,beta-amyrin produced much greater antinociceptive effect at 100mg/kg in the capsaicin test, which was also naloxone-sensitive. These results provide first time evidence to show that alpha,beta-amyrin attenuates orofacial pain at least, in part, through a peripheral opioid mechanism but warrants further detailed study for its utility in painful orofacial pathologies.

Anti-inflammatory effect of alpha, beta-Amyrin, a pentacyclic triterpene from Protium heptaphyllum in rat model of acute periodontitis.

This study was aimed to evaluate the anti-inflammatory potential of triterpene alpha, beta-amyrin in rats on acute phase periodontitis. Periodontitis was induced by ligature placement around the maxillary right second molar tooth. Rats (n = 8/group) were pretreated with alpha, beta-amyrin (5 and 10 mg/kg, p. o.), two hours before the induction of periodontal inflammation. Sham-operated and positive controls (lumiracoxib and dexamethasone) were included. Six hours later, plasma levels of TNF-alpha were analysed. Rats were sacrificed at 24 h, and the gingival tissue analysed for myeloperoxidase (MPO) and thiobarbituric acid-reactive substances (TBARS), as measures of neutrophil influx and lipid-peroxidation, respectively alpha, beta-Amyrin as well as dexamethasone significantly inhibited the periodontitis-associated increases of TNF-alpha, and the gingival MPO and TBARS. alpha, beta-Amyrin effect was more prominent at 5 mg/kg. Lumiracoxib manifested varied influence on the studied parameters. These results provide evidence to show that alpha, beta-Amyrin retards acute inflammation in rat model of periodontitis and warrant further study on its efficacy to prevent chronic periodontitis-associated bone loss.

Review series: the politics of TB: the politics, economics and impact of directly observed treatment (DOT) in India.

India harbors approximately one-third of the world's tuberculosis cases. The disease being multi-factorial; various political, social and economic factors play pivotal roles in causation and control. The country's policy-makers, via the Revised National Tuberculosis Programme (RNTCP), have embraced DOTS, i.e. Directly Observed Treatment; short course, as a means of combating the disease. Today, a decade after being launched, the DOTS programme in India is the largest in the world. The achievements of the programme have been significant in reaching out to the millions and having impressive cure rates, but the disease is far from eradicated. Social taboos, economic obstacles, and deficient infrastructure are impediments that hamper the success of the programme. With multidrug-resistant TB and HIV co-infection complicating the management of TB; the task has become more daunting. In a country as heterogeneous as India, novel holistic strategies that address individual needs will have to be developed to successfully curb the spread of the disease in the future.

NK cells, displaying early activation, cytotoxicity and adhesion molecules, are associated with mild dengue disease.

During the innate immune response against infections, Natural Killer (NK) cells are as important effector cells as are Cytotoxic T lymphocytes (CTL) generated after antigenic stimulation in the adaptative response. NK cells increase in numbers, after viral infection or vaccination. We investigated the NK cell and CD8 T lymphocyte status in 55 dengue infected patients. The NK (CD56+CD3-) and CD56+ T cell (CD56+CD3+) rates rise during the acute phase of disease. The majority of NK cells from dengue patients display early markers for activation (CD69, HLA-DR, and CD38) and cell adhesion molecules (CD44, CD11a) during the acute phase of disease. The intracellular cytotoxic granule, TIA-1, is also up-regulated early in NK cells. Most of these markers appear also on CD8+ T lymphocytes but during the late acute phase. Circulating IL-15 is elevated in a significant number of patients during early acute infection and its values were statistically correlated with NK frequencies and cytotoxic markers on NKs. We have therefore shown that dengue virus infection is very likely stimulating a cytotoxic response that may be efficient in controlling the virus in synergism with CD8+ T lymphocytes. Interestingly, the heightened CD56+CD3-, CD56+CD3+, CD56+TIA-1+ and CD56+CD11a+ cell rates are associated with mild dengue clinical manifestations and might indicate a good prognosis of the disease.

Oral and facial inclusion cysts in newborns.

This study assessed the frequency of milia, alveolar and palatal cysts prevalence in newborns as well as the time for complete involution of the most common reported inclusion cyst. Milia, alveolar and palatal cysts were observed in respectively, 31.4%, 28.2% and 78.8% of the newborns. Those that presented milia were 1.2 times more likely to present alveolar cysts. Mean time for complete involution of palatal cysts was 4.5 weeks.

CC-chemokine receptors: a potential therapeutic target for Trypanosoma cruzi-elicited myocarditis.

The comprehension of the pathogenesis of Trypanosoma cruzi-elicited myocarditis is crucial to delineate new therapeutic strategies aiming to ameliorate the inflammation that leads to heart dysfunction, without hampering parasite control. The augmented expression of CCL5/RANTES and CCL3/MIP-1alpha, and their receptor CCR5, in the heart of T. cruzi-infected mice suggests a role for CC-chemokines and their receptors in the pathogenesis of T. cruzi-elicited myocarditis. Herein, we discuss our recent results using a CC-chemokine receptor inhibitor (Met-RANTES), showing the participation of CC-chemokines in T. cruzi infection and unraveling CC-chemokine receptors as an attractive therapeutic target for further evaluation in Chagas disease.

CC-chemokine receptors: a potential therapeutic target for Trypanosoma cruzi-elicited myocarditis

The comprehension of the pathogenesis of Trypanosoma cruzi-elicited myocarditis is crucial to delineate new therapeutic strategies aiming to ameliorate the inflammation that leads to heart dysfunction, without hampering parasite control. The augmented expression of CCL5/RANTES and CCL3/MIP-1alpha, and their receptor CCR5, in the heart of T. cruzi-infected mice suggests a role for CC-chemokines and their receptors in the pathogenesis of T. cruzi-elicited myocarditis. Herein, we discuss our recent results using a CC-chemokine receptor inhibitor (Met-RANTES), showing the participation of CC-chemokines in T. cruzi infection and unraveling CC-chemokine receptors as an attractive therapeutic target for further evaluation in Chagas disease.

Pathogenicity of Bacillus thuringiensis isolated from two species of Acromyrmex (Hymenoptera, Formicidae).

The control of Acromyrmex leaf-cutting ants is necessary due to the severe damage they cause to diverse crops. A possibility was to control them using the bacterium Bacillus thuringiensis (Bt) that characteristically produces insecticidal crystal proteins (ICPs). The ICPs have been effective in controlling lepidopterans, dipterans, and coleopterans, but their action against hymenopterans is unknown. This paper describes an attempt to isolate Bt from ants of two Acromyrmex species, to evaluate its pathogenicity towards these ants, and to test isolates by PCR. Bacterial isolates of Bt obtained from A. crassispinus and A. lundi have been assayed against A. lundi in the laboratory. The bioassays were carried out in BOD at 25 degrees C, with a 12-hour photoperiod, until the seventh day after treatment. The Bt isolates obtained were submitted to total DNA extraction and tested by PCR with primers specific to cry genes. The results showed Bt presence in 40% of the assessed samples. The data from the in vivo assays showed a mortality rate higher than 50% in the target population, with the Bt HA48 isolate causing 100% of corrected mortality. The PCR results of Bt isolates showed a magnification of DNA fragments relative to cry1 genes in 22% of the isolates, and cry9 in 67%. Cry2, cry3, cry7, and cry8 genes were not detected in the tested samples, and 22% had no magnified DNA fragments corresponding to the assessed cry genes. The results are promising not only regarding allele identification in new isolates, but also fort the assays aimed at determining the Bt HA48 LC50's, which can eventually be applied in controlling of Acromyrmex leaf-cutting ants.

Pathogenicity of Bacillus thuringiensis isolated from two species of Acromyrmex (Hymenoptera, Formicidae)

The control of Acromyrmex leaf-cutting ants is necessary due to the severe damage they cause to diverse crops. A possibility was to control them using the bacterium Bacillus thuringiensis (Bt) that characteristically produces insecticidal crystal proteins (ICPs). The ICPs have been effective in controlling lepidopterans, dipterans, and coleopterans, but their action against hymenopterans is unknown. This paper describes an attempt to isolate Bt from ants of two Acromyrmex species, to evaluate its pathogenicity towards these ants, and to test isolates by PCR. Bacterial isolates of Bt obtained from A. crassispinus and A. lundi have been assayed against A. lundi in the laboratory. The bioassays were carried out in BOD at 25ºC, with a 12-hour photoperiod, until the seventh day after treatment. The Bt isolates obtained were submitted to total DNA extraction and tested by PCR with primers specific to cry genes. The results showed Bt presence in 40 percent of the assessed samples. The data from the in vivo assays showed a mortality rate higher than 50 percent in the target population, with the Bt HA48 isolate causing 100 percent of corrected mortality. The PCR results of Bt isolates showed a magnification of DNA fragments relative to cry1 genes in 22 percent of the isolates, and cry9 in 67 percent. Cry2, cry3, cry7, and cry8 genes were not detected in the tested samples, and 22 percent had no magnified DNA fragments corresponding to the assessed cry genes. The results are promising not only regarding allele identification in new isolates, but also fort the assays aimed at determining the Bt HA48 LC50's, which can eventually be applied in controlling of Acromyrmex leaf-cutting ants

Detection of circulant tumor necrosis factor-alpha, soluble tumor necrosis factor p75 and interferon-gamma in Brazilian patients with dengue fever and dengue hemorrhagic fever.

Pro-inflammatory cytokines are believed to play an important role in the pathogenesis of dengue infection. This study reports cytokine levels in a total of 54 patients examined in Recife, State of Pernambuco, Brazil. Five out of eight patients who had hemorrhagic manifestations presented tumor necrosis factor-alpha (TNF-alpha) levels in sera which were statistically higher than those recorded for controls. In contrast, only one out of 16 patients with mild manifestations had elevated TNF-alpha levels. The levels of interleukin-6 (IL), IL-1beta tested in 24 samples and IL-12 in 30 samples were not significantly increased. Interferon-g was present in 10 out of 30 patients with dengue. The data support the concept that the increased level of TNF-alpha is related to the severity of the disease. Soluble TNF receptor p75 was found in most patients but it is unlikely to be related to severity since it was found with an equivalent frequency and levels in 15 patients with dengue fever and another 15 with dengue hemorrhagic fever.

Interaction of benzocaine with model membranes.

We measured the absorption properties, water solubility and partition coefficients (P) between n-octanol, egg phosphatidylcholine (EPC) liposomes and erythrocyte ghosts/water for benzocaine (BZC), an ester-type always uncharged local anesthetic. The interaction of BZC with EPC liposomes was followed using Electron Paramagnetic Resonance, with spin labels at different positions in the acyl chain (5, 7, 12, 16-doxylstearic acid methyl ester). Changes in lipid organization upon BZC addition allowed the determination of P values, without phase separation. The effect of BZC in decreasing membrane organization (maximum of 11.6% at approx. 0.8:1 BZC:EPC) was compared to those caused by the local anesthetics tetracaine and lidocaine. Hemolytic tests revealed a biphasic (protective/inductive) concentration-dependent hemolytic effect for BZC upon rat erythrocytes, with an effective BZC:lipid molar ratio in the membrane for protection (RePROT), onset of hemolysis (ReSAT) and 100% membrane solubilization (ReSOL) of 1.0:1, 1.1:1 and 1.3:1, respectively. The results presented here reinforce the importance of considering hydrophobic interactions in the interpretation of the effects of anesthetics on membranes.