Evaluation of medullary cytokine expression and clinical and laboratory aspects in severe human visceral leishmaniasis.

BACKGROUND: Visceral leishmaniasis (VL) is a serious public health problem. The factors that can determine whether VL develops and progresses to severe form have not been fully identified, but a specific cellular immune response appears to play a key role. Therefore, understanding immunopathogenesis can be useful in preventing a serious clinical outcome. MATERIALS AND METHODS: Bone marrow samples were collected from patients with severe VL (SVL) or non-severe VL (NSVL). Cytokine levels and parasitic load were analysed by RT-qPCR. There is a statistically significant difference in the leukocyte parameter in patients with SVL and NSVL compared with the control patients (p = .006 and p = .014, respectively). RESULTS: Urea, alanine transaminase and albumin parameters had a significant difference p = .036, p = .039 and p = .017, respectively, between SVL and NSVL. Although high levels of IFN-γ, IL-10, IL-6 and TNF-α were present in all groups of individuals with VL, they were not statistically associated with severity. In patients with active VL, IFN-γ and IL-10 were associated, respectively, with a reduction and increase in the parasite load, strong and significant positive association between IFN-γ and IL-10 (rho = .627 and p = .003). CONCLUSION: This study demonstrates that VL stimulates an non-dichotomized inflammatory response between Th1/Th2 and that bone marrow is an important tissue for immune regulation.

Are oral overuse behaviours associated with painful temporomandibular disorders? A cross-sectional study in Portuguese university students.

BACKGROUND: Many different types of oral overuse behaviours occur frequently in adult populations with painful temporomandibular disorders (TMDs). Less is known regarding these behaviours and their associations with TMDs in university students. OBJECTIVES: Test the association between frequency of different oral overuse behaviours evaluated by the Oral Behaviour Checklist (OBC) and the severity of painful TMDs. METHODS: In this cross-sectional study, 1381 students from 19 universities in the Oporto District, Portugal, completed the Research Diagnostic Criteria for TMD (RDC/TMD) Personal History Questionnaire and the OBC, and they received an RDC/TMD clinical examination. The OBC sum score (ranging from 0 to 84 points) was classified as normal (0 ≤ 16 points), low overuse (17 ≤ 24) or high overuse (≥25). Painful TMD subtypes (myalgia, arthralgia or combined) were identified. Associations were tested using multivariable binary logistic regression models (α = .05), adjusted for age and sex, and referencing the normal parafunction group. RESULTS: University students with high overuse were more likely to have a painful TMD: myalgia (OR = 1.9, 95% CL: 1.3-3.0); arthralgia (OR = 2.2; 95% CL: 1.4-3.4), combined (OR = 5.0; 95% CL: 3.1-8.1). Students with low overuse were more likely to have only the combined painful TMD (OR = 2.4; 95% CL: 1.4-4.0) but not the individual painful disorders. Of the 21 different behaviours, 13 were reported at least 50% of the time. CONCLUSIONS: In this university student sample, oral overuse behaviours are widespread, and their overall extent exhibited a dose-response relationship with respect to severity of painful TMDs based on pain and chronicity. Only some behaviours were independently associated with painful TMDs, suggesting the value of further OBC instrument development.

Relationship Between Running Economy and Kinematic Parameters in Long-Distance Runners.

Pizzuto, F, Fonseca de Oliveira, C, Amorim Soares, TS, Rago, V, Silva, G, and Oliveira, J. Relationship between running economy and kinematic parameters in long-distance runners. J Strength Cond Res 33(7): 1921-1928, 2019-The purpose of this study was to explore the relationship between running economy (RE) and sagittal, frontal, and transverse plane kinematic parameters in long-distance runners. A secondary purpose was to identify the kinematic predictors of RE during running at the lowest RE value, representing an individual's most efficient running intensity. Twenty recreational long-distance runners ran 3 submaximal stages on a treadmill (65, 75, and 85% of velocity at maximum oxygen consumption). Respiratory data were collected using a portable gas analysis system. Kinematics were gathered using passive retroreflective markers and 8 high-resolution infrared cameras to collect the respective trajectories. Hip, knee, and ankle angles at foot strike and stance phase, as well as spatio-temporal parameters were calculated during each gait cycle. Knee flexion/extension range of motion (ROM), knee ab/adduction ROM, and hip ab/adduction ROM during the stance phase of the gait cycle showed positive moderate to large correlations with RE (r ± 90% confidence intervals = 0.51 ± 0.29; 0.49 ± 0.30; 0.53 ± 0.28, respectively). Knee and hip ab/adduction ROMs during the stance phase are predictors of RE, accounting for 44% of RE variance. Therefore, sagittal and frontal plane kinematics affect RE-inducing alterations in running performance. Coaches, athletic trainers, and anyone involved in running training prescription should consider a relationship between these parameters to ensure optimal technique and, consequently, to improve RE in recreational long-distance runners.

Histomorphometric approach to differentiate skin lesions of tuberculoid leprosy from sarcoidosis.

BACKGROUND: More than 200 000 new cases of leprosy are detected worldwide annually. Physicians commonly have difficulty in differentiating tuberculoid form of leprosy (TL) from sarcoidosis' cutaneous manifestation. METHODS: Skin biopsies of 33 patients with TL and 24 with sarcoidosis were reviewed on hematoxylin and eosin- and Gomori-stained sections, in order to find reliable criteria for distinguishing one disease from another. RESULTS: Nine of the 24 features analyzed presented significant predictive value for diagnosis (P < .05). Predominance of tuberculoid granulomas in adnexal and neural distribution, and granulomas replacing the nerves localized within sweat gland glomeruli were predictive to TL diagnosis. For sarcoidosis, dermal fibrosis, back-to-back distribution of the granulomas, presence of atypical giant cells and plasma cells, greater number of conventional giant cells, and spared nerves beside the granuloma were predictive criteria. The median surface density of reticulin fibers was significantly higher in sarcoidosis (3.44) than in TL (2.99). Nonetheless, using logistic regression, this variable did not discriminate between the diseases (P = .096). CONCLUSIONS: Isolated histological features are not fully predictive to differentiate the 2 diseases. However, those with statistical value can assist this distinction in diagnostic practice. Although the results of the analysis of the reticulin fibers density did not tell apart TL from sarcoidosis, they corroborate the idea of fiber fragmentation within tuberculoid leprosy granulomas, reiterating the importance of morphometry in the histological examination.

Acute kinematics changes in marathon runners using different footwear.

The effects of running with or without shoes on injury prevention have been extensively studied, and several investigations have assessed biomechanical differences between them. However, findings are not consensual and further insights on biomechanical load associated with differently shod or barefoot conditions may be needed. This study aimed to observe if habitually shod marathon runners show acute alterations when running barefoot or with minimalist shoes, and to determine whether the running kinematical adaptations of wearing minimalist shoes were similar to barefoot running. Twelve male marathon runners ran on the treadmill at their average marathon pace in different footwear conditions: habitual running shoes, minimalist shoes, and barefoot. High-resolution infrared cameras and visual 3D software were used to assess kinematic data. The following parameters were studied: foot strike angle, cycle time, stance time, normalized stride length, hip, knee, and ankle angular position at initial contact, and their respective range-of-motion (ROM) during stance phase. Contrary to the expectations, it was found that highly trained habitually shod elite marathon runners changed their lower limb kinematic pattern both when running barefoot or wearing minimalist shoes. Minimalist shoes showed a trend towards intermediate biomechanical effects between running with and without shoes.

Cultural equivalence, reliability and utility of the Portuguese version of the Oral Behaviours Checklist.

BACKGROUND: Oral behaviors (OBs) must be assessed as part of a biobehavioural evaluation for patients with temporomandibular disorders (TMD). OBJECTIVES: The aims of this study were: to translate and culturally adapt the Oral Behaviors Checklist (OBC) into Portuguese; to evaluate its basic psychometric properties; and to assess the utility of the Portuguese OBC. METHODS: The Portuguese translation, cross-cultural equivalence, and two studies (Pre-field test, N = 8, and field test, N = 45) of the OBC adhered to official guidelines of the International RDC/TMD Research Consortium (now, INfORM). In the formal test (N = 120) the Portuguese RDC/TMD was used for TMD diagnosis. RESULTS: Translation and cultural equivalence of the OBC into Portuguese was reviewed and approved by an expert panel. In the field test, item agreement between English and Portuguese OBC versions was very good (weighted Kappa ≥ 0.934). Test-retest reliability of the OBCS umScore was excellent intra-class correlation coefficient ((ICC) = 0.998, P < 0.001). As an initial probe into validity using parallel forms, comparison of the sleep bruxism and awake clenching questions included in the RDC/TMD with the corresponding OBC questions yielded excellent (Kappa = 0.932) and very good (k = 0.850), respectively, convergent validity as well as excellent (that is, poor agreement, kappa = 0.013) discriminant validity for these specific OBs. The mean OBCS umScore was significantly lower for the healthy group, compared to the painful TMD group (ANOVA, P = 0.037). CONCLUSIONS: The Portuguese translation of the OBC exhibits cross-cultural equivalence for use in Portugal, and the instrument performs with acceptable psychometric properties. The validity of the parafunction construct requires additional research in both Portuguese and other languages.

HDAC6 inhibition induces mitochondrial fusion, autophagic flux and reduces diffuse mutant huntingtin in striatal neurons.

Striatal neurons are vulnerable to Huntington's disease (HD). Decreased levels of acetylated alpha-tubulin and impaired mitochondrial dynamics, such as reduced motility and excessive fission, are associated with HD; however, it remains unclear whether and how these factors might contribute to the preferential degeneration of striatal neurons. Inhibition of the alpha-tubulin deacetylase HDAC6 has been proposed as a therapeutic strategy for HD, but remains controversial - studies in neurons show improved intracellular transport, whereas studies in cell-lines suggest it may impair autophagosome-lysosome fusion, and reduce clearance of mutant huntingtin (mHtt) and damaged mitochondria (mitophagy). Using primary cultures of rat striatal and cortical neurons, we show that mitochondria are intrinsically less motile and more balanced towards fission in striatal than in cortical neurons. Pharmacological inhibition of the HDAC6 deacetylase activity with tubastatin A (TBA) increased acetylated alpha-tubulin levels, and induced mitochondrial motility and fusion in striatal neurons to levels observed in cortical neurons. Importantly, TBA did not block neuronal autophagosome-lysosome fusion, and did not change mitochondrial DNA levels, suggesting no impairment in autophagy or mitochondrial clearance. Instead, TBA increased autophagic flux and reduced diffuse mHtt in striatal neurons, possibly by promoting transport of initiation factors to sites of autophagosomal biogenesis. This study identifies the pharmacological inhibition of HDAC6 deacetylase activity as a potential strategy to reduce the vulnerability of striatal neurons to HD.

Mitochondrial dynamics and quality control in Huntington's disease.

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by polyglutamine expansion mutations in the huntingtin protein. Despite its ubiquitous distribution, expression of mutant huntingtin (mHtt) is particularly detrimental to medium spiny neurons within the striatum. Mitochondrial dysfunction has been associated with HD pathogenesis. Here we review the current evidence for mHtt-induced abnormalities in mitochondrial dynamics and quality control, with a particular focus on brain and neuronal data pertaining to striatal vulnerability. We address mHtt effects on mitochondrial biogenesis, protein import, complex assembly, fission and fusion, mitochondrial transport, and on the degradation of damaged mitochondria via autophagy (mitophagy). For an integrated perspective on potentially converging pathogenic mechanisms, we also address impaired autophagosomal transport and abnormal mHtt proteostasis in HD.

Bartonella henselae transmission by blood transfusion in mice.

BACKGROUND: Bartonella spp. are neglected fastidious Gram-negative bacilli. We isolated Bartonella henselae from 1.2% of 500 studied blood donors and demonstrated that the bacteria remain viable in red blood cell units after 35 days of experimental infection. Now, we aim to evaluate the possibility of B. henselae transmission by blood transfusion in a mouse model. STUDY DESIGN AND METHODS: Eight BALB/c mice were intraperitoneal inoculated with a 30 µL of suspension with 10(4) CFU/mL of B. henselae and a second group of eight mice were inoculated with saline solution and used as control. After 96 hours of inoculation, the animals were euthanized. We collected blood and tissue samples from skin, liver, and spleen. Thirty microliters of blood from four Bartonella-inoculated animals were transfused into a new group (n = 4). Another group received blood from the control animals. B. henselae infection was investigated by conventional and nested polymerase chain reaction (PCR). RESULTS: Blood samples from all 24 mice were negative by molecular tests though half of the tissue samples were positive by nested PCR in the intraperitoneal Bartonella-investigated animals. Tissues from two of the four mice that received blood transfusions from Bartonella-inoculated animals were also nested PCR positives. CONCLUSIONS: Transmission of B. henselae by transfusion is possible in mice even when donor animals have undetectable bloodstream infection. The impact of human Bartonella sp. transmission through blood transfusion recipients must be evaluated.

Bartonella clarridgeiae bacteremia detected in an asymptomatic blood donor.

Human exposure to Bartonella clarridgeiae has been reported only on the basis of antibody detection. We report for the first time an asymptomatic human blood donor infected with B. clarridgeiae, as documented by enrichment blood culture, PCR, and DNA sequencing.

Scoping Review of Fall Risk Assessment Tools for Women Who Receive Maternity Care.

OBJECTIVE: To identify and describe fall risk assessment tools used for women who receive maternity care. DATA SOURCES: PubMed, CINAHL Complete, MEDLINE Complete, Cochrane Library, Scopus, SciELO, and Repositórios Científicos de Acesso Aberto de Portugal (RCAAP). STUDY SELECTION: We considered reports published until November 28, 2022, that included women during pregnancy, childbirth, or the postpartum period; involved the use of fall risk assessment tools, regardless of context; and were published in English, French, Portuguese, or Spanish. DATA EXTRACTION: We extracted the following data from the included reports: author(s)/year/country, aim/sample, research design/type of report, tool (i.e., the fall risk assessment tool used), findings, reliability, and validity. DATA SYNTHESIS: We found 13 reports in which the authors addressed nine fall risk assessment tools. Seven of these tools were applied during pregnancy (Kyle's tool, Pregnant Women Information Form and Assessment Scale for Risk of Falling in Pregnant Women, Obstetric Fall Risk Assessment System), labor (Obstetric Fall Risk Assessment System), the postpartum period (Cooksey-Post Obstetric Delivery Fall Risk Assessment, Kyle's tool, Risk of Falling in Post-partum Women (SLOPE), Obstetric Fall Risk Assessment System, Post-epidural Fall Risk Assessment Score, and Maternal Fall Risk Assessment Scale). The Dionne's Egress Test and the Motor Strength Scale do not address the characteristics of the women who receive maternity care. Psychometric characteristics were available for the Pregnant Women Information Form and Assessment Scale for Risk of Falling in Pregnant Women, Post-epidural Fall Risk Assessment Score, Maternal Fall Risk Assessment Scale, and Risk of Falling in Post-partum Women. CONCLUSION: Some fall risk assessment tools are used to assess women who receive maternity care without proper validation in this specific population. The use of fall risk assessment tools that are validated for women who receive maternity care may help nurses make clinical judgments when assessing fall risk and implement measures for fall prevention.

3D Resin-coated pressure sensor response for bite force assessment: A pilot study.

Background: Occlusal splints with sensors help in the bruxism diagnosis and monitoring, by recording the patient's bite force. The aim of this study was to evaluate the accuracy of a pressure sensor when it is covered with different thicknesses of a 3D printing resin (Anycubic 405nm Translucent Green UV Resin, Anycubic, UK). Methods: In this preliminary study, the evaluated sensor (FlexiForce A201 Sensor, Tekscan) was firstly calibrated without any type of cover material, and later tested with 3D printing resin with different thicknesses (1 mm, 1.15 mm, 1.4 mm and 1.6 mm). The load tests were performed by a force tester (MultiTest 2.5 dV, Mecmesin). Results: When the pressure sensor was covered with resin of 1mm and 1.6 mm thick specimens, a higher difference was found between the applied load and the corresponding sensor reading. Conclusion: It was concluded that it is possible to use this type of pressure sensor and that it showed better accuracy with the 1.15 mm and 1.4 mm 3D printing resin covering.

The Interconnected Mechanisms of Oxidative Stress and Neuroinflammation in Epilepsy.

One of the most important characteristics of the brain compared to other organs is its elevated metabolic demand. Consequently, neurons consume high quantities of oxygen, generating significant amounts of reactive oxygen species (ROS) as a by-product. These potentially toxic molecules cause oxidative stress (OS) and are associated with many disorders of the nervous system, where pathological processes such as aberrant protein oxidation can ultimately lead to cellular dysfunction and death. Epilepsy, characterized by a long-term predisposition to epileptic seizures, is one of the most common of the neurological disorders associated with OS. Evidence shows that increased neuronal excitability-the hallmark of epilepsy-is accompanied by neuroinflammation and an excessive production of ROS; together, these factors are likely key features of seizure initiation and propagation. This review discusses the role of OS in epilepsy, its connection to neuroinflammation and the impact on synaptic function. Considering that the pharmacological treatment options for epilepsy are limited by the heterogeneity of these disorders, we also introduce the latest advances in anti-epileptic drugs (AEDs) and how they interact with OS. We conclude that OS is intertwined with numerous physiological and molecular mechanisms in epilepsy, although a causal relationship is yet to be established.

Perforin and granzyme B expression in oral and cutaneous lichen planus - a comparative study.

BACKGROUND: Although cutaneous and oral lichen planus (LP) share similar histopathological features, oral LP often follows a recalcitrant course while LP skin lesions tend to be self-limiting. Apoptosis, mediated by cytotoxic T-cells in LP, may be triggered by the release of molecules such as perforin and granzyme B. As variation in clinical behavior can reflect differences in LP immune expression, we studied the role of those cytotoxic molecules in oral and cutaneous LP. METHODS: We analyzed 16 cases of cutaneous LP and 29 of oral LP. The sections were studied on hematoxylin and eosin, CD4, CD8, perforin and granzyme B staining. RESULTS: The mean number of immunostained cells expressing each cytotoxic molecule was significantly higher in oral LP than in cutaneous LP. A higher number of single necrotic keratinocytes (apoptotic bodies) was found in oral LP lesions when compared to cutaneous LP. Only in oral LP lesions, a higher number of CD4-positive cells was found in active lesions when compared to regressive lesions. CONCLUSIONS: Our results confirm increased expression of granzyme B and perforin in oral LP lesions as compared to cutaneous LP. The increased expression suggests a relationship with the clinical behavior of the disease.

Perforin and granzyme B involvement in oral lesions of lichen planus and chronic GVHD.

BACKGROUND: Oral lesions of lichen planus and chronic graft-vs.-host disease (cGVHD) have similar clinical and histological features, but distinct etiology. Apoptosis induced by cytotoxic T lymphocyte has been proposed as a mechanism of keratinocytes death. Cytotoxicity can be mediated by granules containing granzyme B and perforin. Since common features can reflect similarities in immunological mechanisms, we studied the role of those molecules in both diseases. METHODS: We analyzed 29 cases of oral lichen planus and 27 of oral cGVHD. The sections were studied on H&E, perforin and granzyme B staining. RESULTS: The total means (epithelium plus connective tissue number) of the granzyme B- and perforin-positive cells were significantly higher in cGVHD than in oral lichen planus lesions (P<0.05). Also, it was found that the higher the number of perforin+ cells, the higher the number of granzyme-B+ cells in the epithelium and in the connective tissue for both groups (P < 0.05). In oral lichen planus, the number of single apoptotic bodies had a positive correlation with connective tissue granzyme immunostaining and a negative correlation with perforin (P<0.01). On the contrary, in oral cGVHD, the number of apoptotic body clusters presented a positive correlation with connective tissue perforin (P<0.01). CONCLUSIONS: Our findings indicate that apoptosis in oral lichen planus seems to be correlated with granzyme B release, while in oral cGVHD, perforin seems to be more important. Although these diseases present clinical and histological similarities, subtle differences seem to exist in their pathogenetic mechanisms.

Bartonella henselae infection induces a persistent mechanical hypersensitivity in mice.

Bartonella spp. are re-emerging and neglected bacterial pathogens. The natural reservoirs for several species of this genus are domestic animals such as cats and dogs, the most common pets in the USA and Brazil. Some cat studies suggest that the infection is more prevalent in tropical and poverty-stricken areas. These bacteria were associated with a wide spectrum of clinical manifestations: fever of unknown origin, endocarditis, angiomatosis, chronic lymphadenopathy, hepatitis, fatigue, paresthesia and pain. Our group has already demonstrated that B. henselae -infected sickle cell disease mice present with hyperalgesia. We hypothesized that even immunocompetent mice infected by B. henselae would show an increased and persistent mechanical sensitivity. Five ten-week old male BALB/c mice were intraperitoneally inoculated with a 30 µL of suspension containing 10 4 CFU/mL of B. henselae, while five others were inoculated with an equal volume of saline solution. Four days after bacterial inoculation, the mechanical paw withdrawal threshold was measured using von Frey filaments in all animals, for five consecutive days. The infected animals showed hypersensitivity to mechanical stimuli for five consecutive days. The present study has demonstrated that B. henselae infection induces persistent mechanical hypersensitivity, a signal consistent with pain.

Prevalence of Bartonella spp. Infection in Patients with Sickle Cell Disease.

Background: The inherent characteristics of the sickle cell disease (SCD), the most common genetic hematological disorder, increase the propensity of infections. Bartonella spp. are emerging and neglected bacteria. A large spectrum of clinical manifestations has been linked to bartonella bloodstream infection in the last two decades that can cause fatal outcomes, especially in immunodeficient patients. The goal of this study was to evaluate the prevalence of bartonella infection in SCD patients. Materials and Methods: We evaluated Bartonella spp. prevalence in 107 SCD patients. Blood samples and enrichment blood cultures were analyzed by molecular detection of Bartonella spp. DNA. Bartonella DNA was amplified using conventional genus-specific Bartonella PCR which amplifies the Intergenic Transcribed Spacer region and Bartonella henselae-specific nested PCR which amplifies the FtsZ gene. Positive patient DNAs were tested with ssrA conventional PCR. All amplicons were sequenced. Findings: Ten of 107 patients tested positive for B. henselae infection in at least one molecular test. All obtained amplicons were sequenced and similar to B. henselae sequences deposited in GenBank (accession number BX897699). Based on statistical results, bloodstream infection with B. henselae was not associated with animal contact or blood transfusions. Conclusion: We detected B. henselae DNA in 10 (9.3%) SCD studied patients. These patients were notified and treatment was offered to them.

Targeting the proteostasis network in Huntington's disease.

Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by a polyglutamine expansion mutation in the huntingtin protein. Expansions above 40 polyglutamine repeats are invariably fatal, following a symptomatic period characterised by choreiform movements, behavioural abnormalities, and cognitive decline. While mutant huntingtin (mHtt) is widely expressed from early life, most patients with HD present in mid-adulthood, highlighting the role of ageing in disease pathogenesis. mHtt undergoes proteolytic cleavage, misfolding, accumulation, and aggregation into inclusion bodies. The emerging model of HD pathogenesis proposes that the chronic production of misfolded mHtt overwhelms the chaperone machinery, diverting other misfolded clients to the proteasome and the autophagy pathways, ultimately leading to a global collapse of the proteostasis network. Multiple converging hypotheses also implicate ageing and its impact in the dysfunction of organelles as additional contributing factors to the collapse of proteostasis in HD. In particular, mitochondrial function is required to sustain the activity of ATP-dependent chaperones and proteolytic machinery. Recent studies elucidating mitochondria-endoplasmic reticulum interactions and uncovering a dedicated proteostasis machinery in mitochondria, suggest that mitochondria play a more active role in the maintenance of cellular proteostasis than previously thought. The enhancement of cytosolic proteostasis pathways shows promise for HD treatment, protecting cells from the detrimental effects of mHtt accumulation. In this review, we consider how mHtt and its post translational modifications interfere with protein quality control pathways, and how the pharmacological and genetic modulation of components of the proteostasis network impact disease phenotypes in cellular and in vivo HD models.

Optimization of Experimental Settings for the Assessment of Reactive Oxygen Species Production by Human Blood.

The purpose of an experimental design is to improve the productivity of experimentation. It is an efficient procedure for planning experiments, so the data obtained can be analyzed to yield a valid and objective conclusion. This approach has been used as an important tool in the optimization of different analytical approaches. A D-optimal experimental design was used here, for the first time, to optimize the experimental conditions for the detection of reactive oxygen species (ROS) produced by human blood from healthy donors, a biological matrix that better resembles the physiologic environment, following stimulation by a potent inflammatory mediator, phorbol-12-myristate-13-acetate (PMA). For that purpose, different fluorescent probes were used, as 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA), 2-[6-(4'-amino)-phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (APF), and 10-acetyl-3,7-dihydroxyphenoxazine (amplex red). The variables tested were the human blood dilution, and the fluorescent probe and PMA concentrations. The experiments were evaluated using the Response Surface Methodology and the method was validated using specific compounds. This model allowed the search for optimal conditions for a set of responses simultaneously, enabling, from a small number of experiments, the evaluation of the interaction between the variables under study. Moreover, a cellular model was implemented and optimized to detect the production of ROS using a yet nonexplored matrix, which is human blood.

Bartonella henselae Infection in Sickle Cell Disease Mice Is Associated with Hyperalgesia.

BACKGROUND: Sickle cell disease (SCD) is the most prevalent hematologic genetic disorder. Acute vaso-occlusive painful crisis is the hallmark of the disease and may be related to subclinical infections. Bartonellosis, a rare and neglected infection, is caused by Bartonella spp., which can be found in donated blood. These bacteria cause intraerythrocytic and endothelial infection and pain, all of which occur in SCD. It is likely that this infection is transmitted to SCD patients during transfusion from donated blood, leading to pain. We, therefore, evaluated whether Bartonella henselae infection would cause hyperalgesia in mice with SCD. MATERIALS AND METHODS: SCD mice were generated by transplantation of nucleated bone marrow cells harvested from transgenic Berkeley sickle mice into 2-month-old irradiated C57BL/6 mice. We infected four SCD mice by intraperitoneal inoculation with B. henselae, and inoculated four other mice with the same volume of saline. Mechanical hyperalgesia was determined using von Frey monofilaments by two blinded observers. Thereafter, the animals were anesthetized and euthanized to collect blood, liver, and spleen samples to seek B. henselae infection by PCR. FINDINGS: We confirmed the experimental infection in all animals by PCR. Tremors and mechanical hypersensitivity were demonstrated by SCD mice infected with B. henselae infection but not in those receiving saline. CONCLUSION: B. henselae infection may be related to pain and other symptoms in SCD.