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.

False Negative Results in Bartonellosis Diagnosis.

We report a fatal case of Bartonella henselae bacteremic patient. He had negative serology and PCRs from whole blood and liquid culture; only ftsZ nested PCR was positive from the blood liquid culture. The isolate had positive PCRs. When considered, bartonellosis diagnosis can be still challenging because of technical limitations.

Topical essential fatty acid oil on wounds: Local and systemic effects.

BACKGROUND: The use of medicinal plants and their derivatives is increasing, and approximately one-third of all traditional herbal medicines are intended for wound treatment. Natural products used in these treatments include vegetable oils, which are rich in essential fatty acids. Once in contact with an ulcerative surface, the oil reaches the blood and lymphatic vessels, thus eliciting systemic effects. OBJECTIVE: This study evaluated the local and possible systemic effects of essential fatty acids (sunflower oil) applied topically to rat wounds. METHODS: Cutaneous punch wounds (6 mm) were produced on the dorsa of 30 rats. Saline (SS), mineral oil (MO) or essential fatty acid (EFA) solutions were applied topically. Healing was evaluated after 2, 4 and 10 days (n = 5 per group) by visual and histological/morphometric examination, second harmonic generation (SHG) microscopy, and cytokine and growth factor quantification in the scar tissue (real-time PCR) and in serum (ELISA). RESULTS: MO/EFA-treated animals had higher IGF-1, leptin, IL-6 and IFN-γ mRNA expression and lower serum IL-6 levels than the control (SS/MO) animals. SHG analysis showed no difference in collagen density between the animals treated with MO and EFA. CONCLUSION: EFA treatment induces topical (observed by local IGF-1, leptin, IL-6 and IFN-γ production) and systemic effects, lowering IL-6 levels in the serum. As the oil is widely used to shorten ulcer healing time, studies are needed to evaluate the treatment safety and possible undesired effects.

Topical use and systemic action of green and roasted coffee oils and ground oils in a cutaneous incision model in rats (Rattus norvegicus albinus).

INTRODUCTION: Wounds are a common health problem. Coffee is widely consumed and its oil contains essential fatty acids. We evaluated the local (skin) and systemic effects associated with the topical use of coffee oils in rats. METHODS: Punch skin wounds (6 mm) incisions were generated on the backs of 75 rats. Saline (SS), mineral oil (MO), green coffee oil (GCO), roasted coffee oil (RCO), green coffee ground oil (GCGO) or roasted coffee ground oil (RCGO) were topically applied to the wounds. Healing was evaluated by visual and histological/morphometric optical microscopy examination; second harmonics generation (SHG) microscopy, wound tissue q-PCR (values in fold-change) and blood serum (ELISA, values in pg/mL). RESULTS: RCO treated animals presented faster wound healing (0.986 vs. 0.422), higher mRNA expression of IGF-1 (2.78 vs. 1.00, p = 0.01), IL-6 (10.72 vs. 1.00, p = 0.001) and IL-23 (4.10 vs. 1.2, p = 0.05) in early stages of wound healing; higher IL-12 (3.32 vs. 1.00, p = 0.05) in the later stages; and lower serum levels of IFN-γ (11.97 vs. 196.45, p = 0.01). GCO treatment led to higher mRNA expression of IL-6 (day 2: 7.94 vs. 1.00, p = 0.001 and day 4: 6.90 vs. 1.00, p = 0.01) and IL-23 (7.93 vs. 1.20, p = 0.001) in the early stages. The RCO treatment also produced higher serum IFN-α levels throughout the experiment (day 2: 52.53 vs. 21.20; day 4: 46.98 vs.21.56; day 10: 83.61 vs. 25.69, p = 0.05) and lower levels of IL-4 (day 4: 0.9 vs.13.36, p = 0.01), adiponectin (day 10: 8,367.47 vs. 16,526.38, p = 0.001) and IFN-γ (day 4: 43.03 vs.196.45, p = 0.05). The SHG analysis showed a higher collagen density in the RCO and GCO treatments (p = 0.05). CONCLUSION: Topical treatment with coffee oils led to systemic actions and faster wound healing in rats. Further studies should be performed are necessary to assess the safety of topical vegetal oil use for skin lesions.

Efeitos do tratamento tópico com ácido hialurônico 0, 2% em queimadura de segundo grau: um relato de experiência / Effects of topical treatment with hyaluronic acid 0, 2% in second-degree burn: an experience report / Efectos del tratamiento tópico con ácido hialurónico 0, 2% quemadura de segundo grado: un relato de experiencia

OBJETIVO: Testar o uso tópico diário e prolongado na cicatrização de queimaduras em idoso, observando-se os parâmetros: tempo de cicatrização, presença ou ausência de cicatrizes hipertróficas e o efeito estético final resultante do tratamento. RELATO DE CASO: Trata-se de um relato de caso realizado em clínicaescola de uma Faculdade de Enfermagem do interior paulista. O estudo descreve o tratamento empregado em lesão por queimadura de segundo grau em uma paciente de 64 anos, que sofreu queimadura de espessura parcial por escaldadura. Foi realizado registro fotográfico e a avaliação da ferida. Inicialmente, a paciente recebeu o tratamento convencional com sulfadiazina de prata 1% durante sete dias. Como não houve resolução, iniciou-se o tratamento com ácido hialurônico (AH) 0,2% diariamente. Com 14 dias de tratamento, observou-se completa reepitelização. Após 27 dias do início do tratamento, a cicatriz apresentou-se com melhora de hiperpigmentação e não havia sinais de hipertrofia. Não foram observados eventos adversos locais ou sistêmicos durante o período de estudo da lesão. CONCLUSÃO: Os dados permitem concluir que a aplicação tópica de AH 0,2% em queimaduras de paciente idoso contribuiu para acelerar a cicatrização, melhorou a evolução do tratamento e o resultado estético.

OBJECTIVE: To test daily and prolonged topical use in the healing of burns in the elderly, considering the parameters: time of healing, presence or absence of hypertrophic scars and the final aesthetic effect resulting from the treatment. CASE REPORT: This is a case report carried out in the clinical school of a School of Nursing at São Paulo state. The study describes the treatment used in second-degree burn injury in a 64-year-old patient who suffered partial thickness burn by scald. Photographic recording and wound evaluation were performed. Initially, the patient received conventional treatment with 1% silver sulfadiazine for seven days. As there was no resolution, treatment with 0.2% hyaluronic acid (HA) was started daily. At 14 days of treatment, complete re-epithelization was observed. After 27 days, the scar presented with improvement of hyperpigmentation and there were no signs of hypertrophy. No local or systemic adverse events were observed during the study period of the lesion. CONCLUSION: The data allow us to conclude that the topical application of AH 0.2% in burns of elderly patients contributed to accelerate healing, improved treatment evolution and aesthetic result.

Acute and Late Bartonella henselae Murine Model Infection.

Bartonella spp. are fastidious gram-negative neglected bacilli with worldwide distribution. They are able to cause intraerythrocytic and potentially fatal infection. Cats and dogs are reservoirs of some species of these agents. Blood-sucking arthropods are potential vectors. Our aim was to evaluate the blood, skin, liver, and spleen in BALB/c mice by using molecular tests and confocal microscopy to demonstrate Bartonella henselae infection in the bloodstream and organs after 4 and 21 days of intraperitoneally injected bacterial suspension. We demonstrate that the occurrence of infection in organs precedes the detectable infection in blood. Therefore, late manifestation in blood may be another challenge in early detection and diagnosis of B. henselae infection.

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 henselae initial infection of mature human erythrocytes observed in real time using bacterial endogenous fluorescence.

Bartonella henselae is a causative agent of anemia, cat scratch disease, bacillary angiomatosis, recurrent fever, hepatitis, endocarditis, chronic lymphadenopathy, joint and neurological disorders. B. henselae are intra-erythrocytic bacteria. The goal of this study was to visualize the B. henselae invasion into enucleated human red blood cells in real time using bacterium endogenous fluorescence. We took advantage of the unique fluorescence emission spectral profile of the bacteria. We used a linear unmixing approach to separate the fluorescence emission spectra of human erythrocytes from native B. henselae when excited at 488nm. Human blood samples were inoculated with B. henselae and incubated for 60 hours. 3-D live images were captured at select intervals using multi-photon laser scanning microscopy. Uninfected blood samples were also analyzed. This study revealed bacteria entering mature erythrocytes over a 60 hour time period.