Integrative phylogenetic, phylogeographic and morphological characterisation of the Unio crassus species complex reveals cryptic diversity with important conservation implications.

The global decline of freshwater mussels and their crucial ecological services highlight the need to understand their phylogeny, phylogeography and patterns of genetic diversity to guide conservation efforts. Such knowledge is urgently needed for Unio crassus, a highly imperilled species originally widespread throughout Europe and southwest Asia. Recent studies have resurrected several species from synonymy based on mitochondrial data, revealing U. crassus to be a complex of cryptic species. To address long-standing taxonomic uncertainties hindering effective conservation, we integrate morphometric, phylogenetic, and phylogeographic analyses to examine species diversity within the U. crassus complex across its entire range. Phylogenetic analyses were performed using cytochrome c oxidase subunit I (815 specimens from 182 populations) and, for selected specimens, whole mitogenome sequences and Anchored Hybrid Enrichment (AHE) data on âˆ¼ 600 nuclear loci. Mito-nuclear discordance was detected, consistent with mitochondrial DNA gene flow between some species during the Pliocene and Pleistocene. Fossil-calibrated phylogenies based on AHE data support a Mediterranean origin for the U. crassus complex in the Early Miocene. The results of our integrative approach support 12 species in the group: the previously recognised Unio bruguierianus, Unio carneus, Unio crassus, Unio damascensis, Unio ionicus, Unio sesirmensis, and Unio tumidiformis, and the reinstatement of five nominal taxa: Unio desectusstat. rev., Unio gontieriistat. rev., Unio mardinensisstat. rev., Unio nanusstat. rev., and Unio vicariusstat. rev. Morphometric analyses of shell contours reveal important morphospace overlaps among these species, highlighting cryptic, but geographically structured, diversity. The distribution, taxonomy, phylogeography, and conservation of each species are succinctly described.

Selection for winners impacts the endocrine system in the Siamese fighting fish.

In southeast Asia, males of the Siamese fighting fish, Betta splendens, have been selected across centuries for winning paired staged fights and previous work has shown that males from fighter strains are more aggressive than wild-types. This strong directional selection for winners is likely to have targeted aggression-related endocrine systems, and a comparison between fighter and wild-type strains can bring into evidence the key hormones implicated in aggression. Here, we compared the plasma levels of the androgen 11-ketotestosterone (KT) and of the corticosteroid cortisol (F) in F2 males of a fighter and a wild-type strain raised under similar laboratory conditions. We show that F was generally lower in fighter as compared with wild-type males, while no overall differences in KT levels were detected between strains. When presented with a mirror-induced aggressive challenge, post-fight levels of F increased but more significantly so in wild-type males, while KT increased in males of both strains. After the challenge, fighter males had higher levels of KT as compared with wild-type males, while the pattern for F was opposite. As compared with animals in social groups, wild-type males placed under social isolation had lower F levels, while KT decreased for fighters. Taken together, this data suggests that while wild-type males responded to aggression with an increase in circulating levels of both androgens and corticosteroids, males selected for winning fights maintained a blunt F response, increasing only KT levels. These data agree with the hypothesis that a combination of high levels of androgens and low levels of corticosteroids is associated with high aggression. Overall, these results seem to indicate that selection for winning had a stronger impact in the hypothalamus-pituitary-interrenal axis than in the hypothalamus-pituitary-gonadal axis in B. splendens.

Impact of seed-applied fungicide and insecticide on Azospirillum brasilense survival and wheat growth-promoting ability.

The use of Azospirillum brasilense as a crop inoculant has increased in recent years. Thus, the compatibility of the inoculation technology with seed treatments using pesticides needs to be evaluated. In this study, we evaluated the effect of an insecticide and fungicide formulation on A. brasilense strain FP2 population by culturing and culture-independent approaches. In addition, we evaluated the impact of these pesticides on the ability of A. brasilense to promote plant growth by monitoring biometric traits (root and shoot dry mass and length) of wheat grown in Greenhouse conditions. Seed pesticide dressings, mainly fungicide, led to a significant mortality of A. brasilense over time. The ability of A. brasilense to promote wheat growth also decreased due to pesticide treatments combined with sowing delay. Considering that pesticides confer fitness advantages to the wheat in field condition, our results suggest that sowing within the first 4 h after inoculation maintain the beneficial effects of A. brasilense on wheat growth promotion. Furthermore, we conclude that inoculation and treatment of seeds with pesticides may be compatible techniques when carried out immediately before sowing.

Influence of environmental factors on the tropical peatlands diazotrophic communities from the Southern Brazilian Atlantic Rain Forest.

The tropical peatlands of southern Brazil are essential for the maintenance of the Atlantic Rain Forest, one of the 25 hotspots of biodiversity in the world. Although diazotrophic micro-organisms are essential for the maintenance of this nitrogen limited ecosystem, so far studies have focused only on micro-organisms involved in the carbon cycle. In this work, peat samples were collected from three tropical peatland regions during dry and rainy seasons and their chemical and microbial characteristics were evaluated. Our results showed that the structure of the diazotrophic communities in the Brazilian tropical peatlands differs in the evaluated seasons. The abundance of the genus Bradyrhizobium showed to be affected by rainfall and peat pH. Despite the shifts of the nitrogen-fixing population in the tropical peatland caused by seasonality it showed to be constantly dominated by α-Proteobacteria followed by Cyanobacteria. In addition, more than 50% of nifH gene sequences have not been classified, indicating the necessity for more studies in tropical peatland, since the reduction of N supply in the peatlands stimulates the recalcitrant organic matter decomposition performed by peatland micro-organisms, influencing the C stock.

Quantum Nonlinear Optics Based on Two-Dimensional Rydberg Atom Arrays.

We propose the combination of subwavelength, two-dimensional atomic arrays and Rydberg interactions as a powerful platform to realize strong, coherent interactions between individual photons with high fidelity. The atomic spatial ordering guarantees efficient atom-light interactions without the possibility of scattering light into unwanted directions, allowing the array to act as a perfect mirror for individual photons. In turn, Rydberg interactions enable single photons to alter the optical response of the array within a potentially large blockade radius R_{b}, which can effectively punch a large "hole" for subsequent photons. We show that such a system enables a coherent photon-photon gate or switch, with a significantly better error scaling (∼R_{b}^{-4}) than in a disordered ensemble. We also investigate the optical properties of the system in the limit of strong input intensities and show that this many-body quantum driven dissipative system can be modeled well by a semiclassical model based on holes punched in a classical mirror.

Decidual NK cell-derived conditioned medium from miscarriages affects endometrial stromal cell decidualisation: endocannabinoid anandamide and tumour necrosis factor-α crosstalk.

STUDY QUESTION: What are the effects of endocannabinoid anandamide (AEA) in uterine natural killer (unK) cells from miscarriage decidua, regarding their cytokine profile and endometrial stromal cell (ESC) crosstalk? SUMMARY ANSWER: uNK-conditioned media from miscarriage samples present high TNF-α levels which inhibit ESC decidualisation. WHAT IS KNOWN ALREADY: AEA plasma levels are higher in women who have suffered a miscarriage. Moreover, AEA inhibits ESC proliferation and differentiation, although the levels and impact on the uNK cell cytokine profile at the feto-maternal interface remain elusive. STUDY DESIGN, SIZE, DURATION: This laboratory-based study used human primary uNK cells which were isolated from first-trimester decidua (gestational age, 5-12 weeks) derived from 8 women with elective pregnancy termination and 18 women who suffered a miscarriage. PARTICIPANTS/MATERIALS, SETTING, METHODS: The first-trimester placental tissues were assayed for AEA levels by UPLC-MS/MS and respective enzymatic profile by western blot. The uNK cells were isolated and maintained in culture. The expression of angiogenic markers in uNK cells was examined by quantitative PCR (qPCR). The uNK-conditioned medium was analysed for IFN-γ, TNF-α and IL-10 production by enzyme-linked immunosorbent assay, and the impact on ESC differentiation was assessed by measuring decidual markers Prl, Igfbp-1 and Fox01 mRNA expression using qPCR. MAIN RESULTS AND THE ROLE OF CHANCE: AEA levels were higher in miscarriage decidua compared with decidua from elective terminations. The uNK cell-conditioned medium from the miscarriage samples exhibited high TNF-α levels and interfered with the decidualisation of ESCs. Exacerbated inflammation and elevated TNF-α levels at the feto-maternal interface may trigger AEA signalling pathways that, in turn, may impact decidualisation and the angiogenic ability of uNK cells. LARGE-SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Primary uNK cell responses are based on a simple in vitro model. Thus, in complex microenvironments, such as the feto-maternal interface, the mechanisms may not be exactly the same. Also, the inflammatory events of miscarriage that, in this study, have happened prior to processing of the samples may cause different responses to that observed. In addition, the magnitude of the inflammatory response, required to trigger the AEA pathways that impact decidualisation and the uNK angiogenic ability in vivo, is still unclear. WIDER IMPLICATIONS OF THE FINDINGS: The endocannabinoid AEA is a modulator of reproductive competence. AEA not only may contribute to neuroendocrine homeostasis but also can take part in uterine changes occurring during early pregnancy. STUDY FUNDING/COMPETING INTEREST(S): The work was supported by UID/MULTI/04378/2019 with funding from Fundação para a Ciência e a Tecnologia (FCT)/MCTES through national funds and PORTUGAL 2020 Partnership Agreement, NORTE-01-0145-FEDER-000024. S.C. Cunha acknowledges FCT for the IF/01616/2015 contract. There are no conflicts of interest.

Artificial selection for male winners in the Siamese fighting fish Betta splendens correlates with high female aggression.

In Southeast Asia, males of the Siamese fighting fish Betta splendens have been selected across centuries for paired-staged fights. During the selection process, matched for size males fight in a small tank until the contest is resolved. Breeders discard losing batches and reproduce winner batches with the aim of increasing fight performance. We assessed the results of this long-term selection process by comparing under standard laboratory conditions male and female aggressive behaviour of one strain selected for staged fights ("fighters") and one strain of wild-types. The aggressive response of adult fish was tested against their mirror image or a size-matched conspecific. Fighter males were more aggressive than wild-type males for all measured behaviours. Differences were not only quantitative but the pattern of fight display was also divergent. Fighter males had an overall higher swimming activity, performing frequent fast strikes in the direction of the intruder and displaying from a distance. Wild-type males were less active and exhibited aggressive displays mostly in close proximity to the stimuli. Females of the fighter strain, which are not used for fights, were also more aggressive than wild-type females. Aggressive behaviours were correlated across male and female fighter siblings, suggesting common genetic and physiological mechanisms to male and female aggression in this species. The study further shows that results were largely independent of the stimulus type, with the mirror test inducing similar and less variable responses than the live conspecific presentation. These results suggest that selection for male winners co-selected for high-frequency and metabolic demanding aggressive display in males and also enhanced female aggression, opening a wide range of testable hypothesis about the ultimate and proximate mechanisms of male and female aggression in B. splendens.

Effects of cannabis tetrahydrocannabinol on endocannabinoid homeostasis in human placenta.

Cannabis use has become a hot topic in several countries due to the debate about its legalization for medical purposes. However, data are limited regarding adverse events, safety and potential impact on reproductive health. Cannabis consumption during pregnancy has been associated with gestational disorders such as preterm birth, intrauterine growth restriction, low birth weight and increased risk of miscarriage, though the underlying biochemical mechanisms are still unknown. Given that the endocannabinoid system (ECS) is involved in several reproductive processes, we tested the hypothesis that the negative outcomes may result from the impact on the ECS homeostasis caused by the main psychoactive compound of cannabis, Δ9-tetrahydrocannabinol (THC). We demonstrate that THC (10-40 µM) impairs placental endocannabinoid system by disrupting the endocannabinoid anandamide (AEA) levels and the expression of AEA synthetic and degrading enzymes N-arachidonoylphosphatidylethanolamine-specific phospholipase D (NAPE-PLD) and fatty acid amide hydrolase (FAAH), respectively. Although, no alterations in cannabinoid receptors CB1 and CB2 expression were observed. Thus, long-term local AEA levels are associated with a shift in the enzymatic profile to re-establish ECS homeostasis. In chronic cannabis users, high AEA levels in placenta may disturb the delicate balance of trophoblast cells turnover leading to alterations in normal placental development and foetal growth.

Astaxanthin counteracts the effects of heat shock on the maturation of bovine oocytes.

The cellular mechanisms induced by elevated temperature on oocytes are not fully understood. However, there is evidence that some of the deleterious effects of heat shock are mediated by a heat-induced increase in reactive oxygen species (ROS). In this context, carotenoid antioxidants might have a thermoprotective effect. Therefore, the objective of this study was to determine the role of astaxanthin (AST) on oocyte ROS production and on the redox profile and developmental competency of cumulus-oocyte complexes (COCs) after 14h heat shock (41°C) during in vitro maturation (IVM). Exposure of oocytes to heat shock during IVM increased ROS and reduced the ability of the oocyte to cleave and develop to the blastocyst stage. However, 12.5 and 25nM astaxanthin rescued these negative effects of heat shock; astaxanthin counteracted the heat shock-induced increase in ROS and restored oocyte developmental competency. There was no effect of astaxanthin on maturation medium lipid peroxidation or on glutathione peroxidase and catalase activity in oocytes and cumulus cells. However, astaxanthin stimulated superoxide dismutase (SOD) activity in heat-shocked cumulus cells. In conclusion, direct heat shock reduced oocyte competence, which was restored by astaxanthin, possibly through regulation of ROS and SOD activity in oocytes and COCs.

Crystal clear: Vibrational spectroscopy reveals intrabone, intraskeleton, and interskeleton variation in human bones.

OBJECTIVES: Vibrational spectroscopy is a valuable tool for the study of burned skeletal remains. Nonetheless, most investigations have been focused on a limited number of samples as well as on faunal bones rather than human bones. Conclusions based on those investigations may lack representativeness, namely about the intrabone, intra- and interskeleton variability of several chemometric indices. We aimed to investigate this issue on a large sample of human bones. MATERIAL AND METHODS: Powder samples were collected from 168 bones from four human skeletons. The sampling targeted 47 long bones, 72 short bones, and 49 tarsal bones as well as different bone regions in a total of 638 powder samples. Bones were experimentally burned in an electric muffle furnace for two hours to maximum temperatures ranging from 400°C to 1000°C. Another 623 burned samples were then collected totaling 1261 samples subjected to FTIR-ATR analysis. The CI, BPI, C/C, and OH/P indices were calculated. RESULTS: An important intrabone, intra- and interskeleton variation was observed, especially for the BPI. The CI, C/C, and OH/P indices revealed much less variation so site-specific sampling may not be as critical in these cases. Clear differences between our results and those from previous investigations were observed, namely on the temperature increment evolution of the CI and C/C indices. DISCUSSION: The relatively large heterogeneity, especially at the intrabone level, is possibly the consequence of microstructural bone differences. The dissimilarities observed between our investigation and other published studies are probably due to the fact that the samples used here came from human rather than faunal bones. Also, our samples were buried previously to the experimental burning so this may also partly explain our contrasting results, since previous research was mostly performed on fresh bone. Future inferences based on vibrational spectroscopy analyses should take into account the possible effect of all these sources.

Short communication: Extended-spectrum AmpC-producing Escherichia coli from milk and feces in dairy farms in Brazil.

The AmpC enzyme is normally expressed constitutively in Escherichia coli, and its overproduction confers resistance to cefoxitin. A newly reported AmpC, the extended-spectrum AmpC (ESAC), is related to resistance to cefepime, a fourth-generation cephalosporin. This enzyme presents more flexibility in the active site due to insertions, replacements, and deletions on AA sequences. Many isolates producing ESAC were reported in human clinical isolates, but E. coli ESAC producers were reported in animals only in France. The animal E. coli strains can produce this enzyme and possibly disseminate it to human and production environments. In our study, 3 strains of E. coli from milk and feces bovine samples, collected in Rio de Janeiro, Brazil, were suspected to produce ESAC. After excluding other mechanisms of resistance, the gene was sequenced to verify ESAC characteristics. These strains presented replacement of AA in omega and R2 loops, suggesting ESAC production. This is the first report to study ESAC E. coli in dairy farms in Brazil.

Dead weight: Validation of mass regression equations on experimentally burned skeletal remains to assess skeleton completeness.

In very fragmentary remains, the thorough inventory of skeletal elements is often impossible to accomplish. Mass has been used instead to assess the completeness of the skeleton. Two different mass-based methods of assessing skeleton completeness were tested on a sample of experimentally burned skeletons with the objective of determining which of them is more reliable. The first method was based on a simple comparison of the mass of each individual skeleton with previously published mass references. The second method was based on mass linear regressions from individual bones to estimate complete skeleton mass. The clavicle, humerus, femur, patella, metacarpal, metatarsal and tarsal bones were used. The sample was composed of 20 experimentally burned skeletons from 10 males and 10 females with ages-at-death between 68 and 90years old. Results demonstrated that the regression approach is more objective and more reliable than the reference comparison approach even though not all bones provided satisfactory estimations of the complete skeleton mass. The femur, humerus and patella provided the best performances among the individual bones. The estimations based on the latter had root mean squared errors (RMSE) smaller than 300g. Results demonstrated that the regression approach is quite promising although the patella was the only reasonable predictor expected to survive sufficiently intact to a burning event at high temperatures. The mass comparison approach has the advantage of not depending on the preservation of individual bones. Whenever bones are intact though, the application of mass regressions should be preferentially used because it is less subjective.

Obesity as a risk factor for temporomandibular disorders.

We conducted a clinical cross-sectional study to evaluate the association between obesity and the presence of painful temporomandibular disorders (TMD), controlling for age, gender, presence of migraine, depression, non-specific somatic symptoms and obstructive sleep apnoea syndrome (OSAS) in an adult population. A total of 299 individuals (76·6% women) with a mean age of 36·8 ± 12·8 years were evaluated. TMD were classified using the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Depression and non-specific somatic symptoms were scored by the Symptom Checklist-90, while pain and disability was rated by the Graded Chronic Pain Scale. Bioimpedanciometry (BIA) was used to assess obesity through total body fat percentage. Migraine was diagnosed according to the criteria of the International Classification of Headache Disorders-2 (ICHD-2). OSAS was classified according to the Berlin Questionnaire. We performed univariate and multivariate models, chi-square tests and odds ratios (OR) with 95% confidence intervals (CI). In the single regression analysis, TMD-pain was significantly associated with total body fat percentage (P = 0·01). In the multivariate analysis, migraine, age, non-specific somatic symptoms and OSAS showed to be stronger predictors of TMD-pain, and obesity did not retain in the regression model. The initial association found between obesity and TMD-pain is lost when it was corrected for gender, migraine, non-specific somatic symptoms and OSAS.

Nanoparticles enhance the ability of human neutrophils to exert phagocytosis by a Syk-dependent mechanism.

BACKGROUND: Some reports indicate that NPs are ingested by cells via different mechanisms, including phagocytosis. In contrast, the direct role of NPs on the phagocytic process is not well documented. The aim of this study was to determine if titanium dioxide (TiO(2)), zinc oxide (ZnO) and cerium dioxide (CeO(2)) NPs, could alter the ability of neutrophils to exert phagocytosis. METHODS: Freshly isolated human neutrophils were incubated with NPs and their ability to phagocytose opsonized sheep red blood cells (SRBCs) or fluorescent latex beads (LBs) was assessed by optical and fluorescence microscopy, respectively. Syk activation was assessed by western blot experiments and a pharmacological approach with piceatannol, a Syk inhibitor, was used to determine its role in NPs-induced neutrophils. The cytokine granulocyte macrophage-colony stimulating factor (GM-CSF) was used as a positive control. RESULTS: All tested NPs enhanced the ability of neutrophil to phagocytose SRBCs and LBs. Syk was activated in NPs-induced neutrophils as evidenced by its increased tyrosine phosphorylation level vs controls and the ability of NPs-induced phagocytosis was reversed by piceatannol. CONCLUSIONS: We found that the tested NPs enhanced phagocytosis, although at different degree, and this occurred by a Syk-dependent mechanism. GENERAL SIGNIFICANCE: This is the first study demonstrating that NPs, by themselves, can directly enhance FcR-mediated (opsonized SRBCs) and complement-mediated (LBs) phagocytosis. Moreover, as part of their mode of action, we determined that NPs can act similarly to GM-CSF leading to Syk activation involved in phagocytosis. This has to be taken under consideration for future nanobiology and nanomedicine studies.

Effects of doxorubicin administration on bone strength and quality in sedentary and physically active Wistar rats.

Doxorubicin (DOX) is used in pediatric cancer treatment. This study assessed the effects of 7 weeks of DOX and 10-week recovery on bone quality and biomechanical properties in sedentary and exercised Wistar rats. DOX decreases femur diaphysis radial growth and biomechanical properties. Some of these DOX effects were aggravated by exercise. INTRODUCTION: Bone growth in pre-pubertal years critically influences adult fracture risk. DOX is widely used in the treatment of pediatric cancers, but there is limited evidence on its potential negative effects on bone growth. Exercise improves bone growth in children, but there is no evidence if it protects against DOX-induced bone toxicity. This study investigates the early and intermediate effects of a 7-week course of DOX on bone histomorphometry and strength in sedentary and exercised growing animal models. METHODS: Sixty-eight male Wistar rats (8 weeks) were treated with DOX (2 mg kg-1) or vehicle for 7 weeks and afterward housed in standard cages or in cages with a running wheel and killed 2 or 10 weeks after last DOX administration. Femurs and blood were collected for assaying geometry, trabecular microarchitecture (histology), biomechanical properties (three-point bending and shearing of the femoral neck), bone calcium content and density (atomic absorption spectroscopy), and bone turnover markers (ELISA). RESULTS: DOX treatment reduced the femur diaphysis radial growth, with DOX-treated animals having a lower tissue area, cortical area, cortical thickness, and moment of inertia. DOX also decreased distal femur trabecular bone volume and trabecular number and increased trabecular separation. Femur diaphysis stiffness and maximum load were also reduced in past DOX-treated animals. Exercise was shown to worsen the effects of past DOX treatment on the femur diaphysis mechanical properties. CONCLUSION: DOX negatively affects bone geometry, trabecular microarchitecture, and femur mechanical properties in growing Wistar rats. Exercise further aggravates the detrimental effects of past DOX treatment on bone mechanical properties.

Isolation and molecular characterization of Leptospira borgpetersenii serovar Hardjo strain Hardjobovis in the urine of naturally infected cattle in Brazil.

Most epidemiologic studies on bovine leptospirosis are based on serological tests that use antibodies against several serotypes, including the serovar Hardjo, which is widespread and considered to be the most adapted to bovine hosts. However, using only serological studies is not sufficient to identify and distinguish species of leptospires. The aim of this study was report the first isolation in Brazil of two strains serovar Hardjo obtained in urine samples from naturally infected cows in a small Brazilian dairy herd and find the genetic species and consequently the type strain Hardjobovis by molecular characterization. Fifteen dairy cows with a history of reproductive failure, such as abortion and infertility, were selected. Urine samples obtained from each animal were immediately seeded in tubes containing Ellinghausen-McCullough-Johnson-Harris culture medium. The identification of the isolates was performed by Multilocus variable-number tandem-repeat analysis (MLVA) technique and phylogenetic analysis of partial sequence of gene sec Y. From the 15 urine samples evaluated, two Leptospira were found and identified as the Londrina 49 and Londrina 54 strains. The MLVA profiles and sequencing of gene sec Y characterized the isolates as L. borgpetersenii serovar Hardjo strain Hadjobovis because it has different genetic pattern of Leptospira interrogans serovar Hardjo strain Hardjoprajitno. Therefore, more studies are needed including isolation and molecular characterization from regional strains to obtain a better knowledge about epidemiology of serovar Hardjo in bovine which may assist in future strategies of prevention and control of bovine leptospirosis.

Headaches and myofascial temporomandibular disorders: overlapping entities, separate managements?

There are relevant clinical overlaps between some of the painful temporomandibular disorders (TMD) and headache conditions that may hamper the diagnostic process and treatment. A non-systematic search for studies on the relationship between TMD and headaches was carried out in the following databases: PubMed, Cochrane Library and Embase. Important pain mechanisms contributing to the close association and complex relationship between TMD and headache disorders are as follows: processes of peripheral and central sensitisation which take place in similar anatomical areas, the possible impairment of the descending modulatory pain pathways and the processes of referred pain. In addition, the clinical examination does not always provide distinguishing information to differentiate between headaches and TMD. So, considering the pathophysiology and the clinical presentation of some types of headache and myofascial TMD, such overlap can be considered not only a matter of comorbid relationship, but rather a question of disorders where the distinction lines are sometimes hard to identify. These concerns are certainly reflected in the current classification systems of both TMD and headache where the clinical consequences of diagnosis such as headache attributed to or associated with TMD are uncertain. There are several similarities in terms of therapeutic strategies used to manage myofascial TMD and headaches. Considering all these possible levels of interaction, we reinforce the recommendation for multidisciplinary approaches, by a team of oro-facial pain specialists and a neurologist (headache specialist), to attain the most precise differential diagnosis and initiate the best and most efficient treatment.

DETECTION OF CTX-M GENE ANTIBIOTICS RESISTANCE IN KLEBSIELLA PNEUMONIA ISOLATES OF HOSPITALS IN ADJARA (GEORGIA).

Research describing the epidemiology of antibiotic resistant microbes is vital to the proactive development of new antimicrobial agents. In the last years, CTX-M extended-spectrum ß-lactamases (ESBLs) have emerged worldwide and have replaced classical TEM and SHV-type ESBLs in many countries. CTX-M-15 is currently the most frequent, with a pandemic distribution, and its rapid spread is facilitated by incorporation of resistance genes in mobile genetic elements. The ESBL is efficacious in Gram-negative bacteria and thus closely associated with nosocomial environments, often colonizing the intestines, particularly in older and dependent patients. Little is known about the CTX-M ESBLs among Klebsiella pneumonia in Adjara. Our paper describes the detected and characterized ESBLs among Klebsiella pneumonia isolates from patients in two different hospitals in Adjara.

Estimation of the pre-burning condition of human remains in forensic contexts.

The determination of the original condition of human remains prior to burning is critical since it may facilitate the reconstruction of circumstances surrounding death in forensic cases. Although the use of heat-induced bone changes is not a completely reliable proxy for determining pre-burning conditions, it is not completely devoid of potential, as we can observe a clear difference in the occurrence of such features between the fleshed and dry bones. In order to quantify this difference and determine its true value for forensic research, the frequencies of heat-induced warping and thumbnail fractures were documented on modern cremations of cadavers from recently deceased individuals and from the cremations of skeletons previously inhumed. The effect of age, sex, time span from death to cremation, duration and temperature of combustion on those frequencies was statistically investigated. Results demonstrated that the heat-induced features were significantly more frequent in the sample of cadavers. In addition, warping was determined to be the most useful indicator of the pre-burning condition of human remains. Temperature of combustion was the only variable having a significant effect on the frequency of both features, suggesting that fluctuation of temperature, along with collagen preservation and recrystallization of the inorganic phase, is paramount for their occurrence. Both warping and thumbnail fractures may eventually be used for the estimation of the pre-burning condition of human remains in lack of other indicators, but their reliability is far from absolute. Ideally, such inference must be supported by other data such as skeletal representation, objects or defleshing marks on the bones.

Broader prevalence of Wolbachia in insects including potential human disease vectors.

Wolbachia are intracellular, maternally transmitted bacteria considered the most abundant endosymbionts found in arthropods. They reproductively manipulate their host in order to increase their chances of being transmitted to the offspring, and currently are being used as a tool to control vector-borne diseases. Studies on distribution of Wolbachia among its arthropod hosts are important both for better understanding why this bacterium is so common, as well as for its potential use as a biological control agent. Here, we studied the incidence of Wolbachia in a broad range of insect species, collected from different regions of Brazil, using three genetic markers (16S rRNA, wsp and ftsZ), which varied in terms of their sensitivity to detect this bacterium. The overall incidence of Wolbachia among species belonging to 58 families and 14 orders was 61.9%. The most common positive insect orders were Coleoptera, Diptera, Hemiptera and Hymenoptera, with Diptera and Hemiptera having the highest numbers of Wolbachia-positive families. They included potential human disease vectors whose infection status has never been reported before. Our study further shows the importance of using quantitative polymerase chain reaction for high-throughput and sensitive Wolbachia screening.