Skin hyperpigmentation following intravenous polymyxin B treatment associated with melanocyte activation and inflammatory process.

What is known and objective Polymyxins were widely used until the 1960s; however, they fell into disfavour owing to their toxicity. The subsequent growth of infections caused by multidrug-resistant Gram-negative bacteria has led to renewed use of this class of antimicrobials in clinical practice. Acquired skin hyperpigmentation (SH) following intravenous polymyxin B treatment has been previously reported, but little is known about its pathogenesis, clinical course and treatment. To improve understanding of these issues, we conducted a prospective study of adult patients receiving intravenous polymyxin B treatment. Methods Patients receiving intravenous polymyxin B treatment were followed throughout the course of treatment. Clinical, dermatoscopic, histologic and immunohistochemical skin properties of patients who presented with SH were studied. Results and discussion Skin hyperpigmentation was noted in 8% of patients (n=20/249); however, clinical, dermatoscopic, histologic and immunohistochemical examinations were performed only in three patients for whom the consent of relatives was obtained. Histologic and immunohistochemical findings showed an abundant melanocyte-pigmented dendritic network. Langerhans cells' hyperplasia and dermal IL-6 overexpression were also found, presumably for an inflammatory process due to polymyxin B use. As polymyxin B causes the release of histamine, which is known for its melanogenic effect, it is possible that skin darkening is associated with this inflammatory mediator. What is new These clinical and dermatoscopic findings contribute to a better understanding of how the pigmentary reaction manifests following intravenous polymyxin B treatment. Conclusion We concluded that hyperpigmentation due to intravenous polymyxin B treatment is associated with an inflammatory process and subsequent melanocyte activation. Although the pigmentary disorder neither influences the outcome of the therapy nor warrants discontinuation of treatment, it nevertheless considerably affects the patient's quality of life.

Lack of ß2-AR improves exercise capacity and skeletal muscle oxidative phenotype in mice.

ß(2)-adrenergic receptor (ß(2)-AR) agonists have been used as ergogenics by athletes involved in training for strength and power in order to increase the muscle mass. Even though anabolic effects of ß(2)-AR activation are highly recognized, less is known about the impact of ß(2)-AR in endurance capacity. We presently used mice lacking ß(2)-AR [ß(2)-knockout (ß(2) KO)] to investigate the role of ß(2)-AR on exercise capacity and skeletal muscle metabolism and phenotype. ß(2) KO mice and their wild-type controls (WT) were studied. Exercise tolerance, skeletal muscle fiber typing, capillary-to-fiber ratio, citrate synthase activity and glycogen content were evaluated. When compared with WT, ß(2) KO mice displayed increased exercise capacity (61%) associated with higher percentage of oxidative fibers (21% and 129% of increase in soleus and plantaris muscles, respectively) and capillarity (31% and 20% of increase in soleus and plantaris muscles, respectively). In addition, ß(2) KO mice presented increased skeletal muscle citrate synthase activity (10%) and succinate dehydrogenase staining. Likewise, glycogen content (53%) and periodic acid-Schiff staining (glycogen staining) were also increased in ß(2) KO skeletal muscle. Altogether, these data provide evidence that disruption of ß(2)-AR improves oxidative metabolism in skeletal muscle of ß(2) KO mice and this is associated with increased exercise capacity.

Exercise training reduces cardiac angiotensin II levels and prevents cardiac dysfunction in a genetic model of sympathetic hyperactivity-induced heart failure in mice.

The role of exercise training (ET) on cardiac renin-angiotensin system (RAS) was investigated in 3-5 month-old mice lacking alpha(2A-) and alpha(2C-)adrenoceptors (alpha(2A)/alpha(2C)ARKO) that present heart failure (HF) and wild type control (WT). ET consisted of 8-week running sessions of 60 min, 5 days/week. In addition, exercise tolerance, cardiac structural and function analysis were made. At 3 months, fractional shortening and exercise tolerance were similar between groups. At 5 months, alpha(2A)/alpha(2C)ARKO mice displayed ventricular dysfunction and fibrosis associated with increased cardiac angiotensin (Ang) II levels (2.9-fold) and increased local angiotensin-converting enzyme activity (ACE 18%). ET decreased alpha(2A)/alpha(2C)ARKO cardiac Ang II levels and ACE activity to age-matched untrained WT mice levels while increased ACE2 expression and prevented exercise intolerance and ventricular dysfunction with little impact on cardiac remodeling. Altogether, these data provide evidence that reduced cardiac RAS explains, at least in part, the beneficial effects of ET on cardiac function in a genetic model of HF.

Changes in the pro-inflammatory cytokine production and peritoneal macrophage function in rats with chronic heart failure.

Chronic heart failure (CHF) is a state of immune activation, and pro-inflammatory cytokines play an important role in its development and progression. Macrophages (Mphis), when activated, are the main source of pro-inflammatory cytokines. We measured interleukin-6 (IL-6), interleukin (IL-1beta) and tumor necrosis factor - alpha (TNF-alpha) production after lipopolysaccharide (LPS)-stimulation, as well as peritoneal Mphis migration, phagocytic capacity, chemotaxis index, and hydrogen peroxide production, in an attempt to clarify the role of this cell in an animal model of CHF. Ligature of the left coronary artery or sham operation was performed in adult Wistar rats. After 12 weeks, resident and total cell number, phagocytic capacity, chemotaxis index, and hydrogen peroxide production in Mphis were significantly higher in CHF than in control rats. The production of IL-6 and TNF- alpha was similarly significantly enhanced in CHF as compared with controls. Mphis obtained from CHF rats were more responsive to LPS, suggesting the existence, in vivo, of possible factor(s) modulating the production of pro-inflammatory cytokines. The results demonstrated that there is modification of peritoneal Mphis function along CHF development, possibly contributing to the pathophysiological process in the establishment of CHF.

The decreased oxygen uptake during progressive exercise in ischemia-induced heart failure is due to reduced cardiac output rate.

We tested the hypothesis that the inability to increase cardiac output during exercise would explain the decreased rate of oxygen uptake (VO2) in recent onset, ischemia-induced heart failure rats. Nine normal control rats and 6 rats with ischemic heart failure were studied. Myocardial infarction was induced by coronary ligation. VO2 was measured during a ramp protocol test on a treadmill using a metabolic mask. Cardiac output was measured with a flow probe placed around the ascending aorta. Left ventricular end-diastolic pressure was higher in ischemic heart failure rats compared with normal control rats (17 +/- 0.4 vs 8 +/- 0.8 mmHg, P = 0.0001). Resting cardiac index (CI) tended to be lower in ischemic heart failure rats (P = 0.07). Resting heart rate (HR) and stroke volume index (SVI) did not differ significantly between ischemic heart failure rats and normal control rats. Peak VO2 was lower in ischemic heart failure rats (73.72 +/- 7.37 vs 109.02 +/- 27.87 mL min(-1) kg(-1), P = 0.005). The VO2 and CI responses during exercise were significantly lower in ischemic heart failure rats than in normal control rats. The temporal response of SVI, but not of HR, was significantly lower in ischemic heart failure rats than in normal control rats. Peak CI, HR, and SVI were lower in ischemic heart failure rats. The reduction in VO2 response during incremental exercise in an ischemic model of heart failure is due to the decreased cardiac output response, largely caused by depressed stroke volume kinetics.

The decreased oxygen uptake during progressive exercise in ischemia-induced heart failure is due to reduced cardiac output rate

We tested the hypothesis that the inability to increase cardiac output during exercise would explain the decreased rate of oxygen uptake (VO2) in recent onset, ischemia-induced heart failure rats. Nine normal control rats and 6 rats with ischemic heart failure were studied. Myocardial infarction was induced by coronary ligation. VO2 was measured during a ramp protocol test on a treadmill using a metabolic mask. Cardiac output was measured with a flow probe placed around the ascending aorta. Left ventricular end-diastolic pressure was higher in ischemic heart failure rats compared with normal control rats (17 ± 0.4 vs 8 ± 0.8 mmHg, P = 0.0001). Resting cardiac index (CI) tended to be lower in ischemic heart failure rats (P = 0.07). Resting heart rate (HR) and stroke volume index (SVI) did not differ significantly between ischemic heart failure rats and normal control rats. Peak VO2 was lower in ischemic heart failure rats (73.72 ± 7.37 vs 109.02 ± 27.87 mL min-1 kg-1, P = 0.005). The VO2 and CI responses during exercise were significantly lower in ischemic heart failure rats than in normal control rats. The temporal response of SVI, but not of HR, was significantly lower in ischemic heart failure rats than in normal control rats. Peak CI, HR, and SVI were lower in ischemic heart failure rats. The reduction in VO2 response during incremental exercise in an ischemic model of heart failure is due to the decreased cardiac output response, largely caused by depressed stroke volume kinetics.

Effects of losartan combined with exercise training in spontaneously hypertensive rats

We investigate whether combined treatment with losartan, an angiotensin II receptor blocker, and exercise training (ET) in spontaneously hypertensive rats (SHR) would have an additive effect in reducing hypertension and improving baroreflex sensitivity when compared with losartan alone. Male SHR (8 weeks old) were assigned to 3 groups: sedentary placebo (SP, N = 16), sedentary under losartan treatment (SL, N = 11; 10 mg kg-1 day-1, by gavage), and ET under losartan treatment (TL, N = 10). ET was performed on a treadmill 5 days/week for 60 min at 50 percent of peak VO2, for 18 weeks. Blood pressure (BP) was measured with a catheter inserted into the carotid artery, and cardiac output with a microprobe placed around the ascending aorta. The baroreflex control of heart rate was assessed by administering increasing doses of phenylephrine and sodium nitroprusside (iv). Losartan significantly reduced mean BP (178 ± 16 vs 132 ± 12 mmHg) and left ventricular hypertrophy (2.9 ± 0.4 vs 2.5 ± 0.2 mg/g), and significantly increased baroreflex bradycardia and tachycardia sensitivity (1.0 ± 0.3 vs 1.7 ± 0.5 and 2.0 ± 0.7 vs 3.2 ± 1.7 bpm/mmHg, respectively) in SL compared with SP. However, losartan combined with ET had no additional effect on BP, baroreflex sensitivity or left ventricular hypertrophy when compared with losartan alone. In conclusion, losartan attenuates hypertension and improves baroreflex sensitivity in SHR. However, ET has no synergistic effect on BP in established hypertension when combined with losartan, at least at the dosage used in this investigation.

Effects of losartan combined with exercise training in spontaneously hypertensive rats.

We investigate whether combined treatment with losartan, an angiotensin II receptor blocker, and exercise training (ET) in spontaneously hypertensive rats (SHR) would have an additive effect in reducing hypertension and improving baroreflex sensitivity when compared with losartan alone. Male SHR (8 weeks old) were assigned to 3 groups: sedentary placebo (SP, N = 16), sedentary under losartan treatment (SL, N = 11; 10 mg kg-1 day-1, by gavage), and ET under losartan treatment (TL, N = 10). ET was performed on a treadmill 5 days/week for 60 min at 50% of peak VO2, for 18 weeks. Blood pressure (BP) was measured with a catheter inserted into the carotid artery, and cardiac output with a microprobe placed around the ascending aorta. The baroreflex control of heart rate was assessed by administering increasing doses of phenylephrine and sodium nitroprusside (iv). Losartan significantly reduced mean BP (178 16 vs 132 12 mmHg) and left ventricular hypertrophy (2.9 0.4 vs 2.5 0.2 mg/g), and significantly increased baroreflex bradycardia and tachycardia sensitivity (1.0 0.3 vs 1.7 0.5 and 2.0 0.7 vs 3.2 1.7 bpm/mmHg, respectively) in SL compared with SP. However, losartan combined with ET had no additional effect on BP, baroreflex sensitivity or left ventricular hypertrophy when compared with losartan alone. In conclusion, losartan attenuates hypertension and improves baroreflex sensitivity in SHR. However, ET has no synergistic effect on BP in established hypertension when combined with losartan, at least at the dosage used in this investigation.

Characterization of novel structures of mannosylinositolphosphorylceramides from the yeast forms of Sporothrix schenckii.

Novel structures of glycoinositolphosphorylceramide (GIPC) from the infective yeast form of Sporothrix schenckii were determined by methylation analysis, mass spectrometry and NMR spectroscopy. The lipid portion was characterized as a ceramide composed of C-18 phytosphingosine N-acylated by either 2-hydroxylignoceric acid (80%), lignoceric (15%) or 2,3-dihydroxylignoceric acids (5%). The ceramide was linked through a phosphodiester to myo-inositol (Ins) which is substituted on position O-6 by an oligomannose chain. GIPC-derived Ins oligomannosides were liberated by ammonolysis and characterized as: Manpalpha1-->6Ins; Manpalpha1-->3Manpalpha1-->6Ins; Manpalpha1-->6Manpalpha1-->3Manpalpha1-->3Manpalpha1-->6Ins; Manpalpha1-->2Manpalpha1-->6Manpalpha1-->3Manpalpha1-->3Manpalpha1-->6Ins. These structures comprise a novel family of fungal GIPC, as they contain the Manpalpha1-->6Ins substructure, which has not previously been characterized unambigously, and may be acylated with a 2,3 dihydroxylignoceric fatty acid, a feature hitherto undescribed in fungal lipids.

Phosphoglucomutase is an in vivo lithium target in yeast.

Lithium is a drug frequently used in the treatment of manic depressive disorder. We have observed that the yeast Saccharomyces cerevisiae is very sensitive to lithium when growing in galactose medium. In this work we show that lithium inhibits with high affinity yeast (IC50 approximately 0.2 mm) and human (IC50 approximately 1.5 mm) phosphoglucomutase, the enzyme that catalyzes the reversible conversion of glucose 1-phosphate to glucose 6-phosphate. Lithium inhibits the rate of fermentation when yeast are grown in galactose and induces accumulation of glucose 1-phosphate and galactose 1-phosphate. Accumulation of these metabolites was also observed when a strain deleted of the two isoforms of phosphoglucomutase was incubated in galactose medium. In glucose-grown cells lithium reduces the steady state levels of UDP-glucose, resulting in a defect on trehalose and glycogen biosynthesis. Lithium acts as a competitive inhibitor of yeast phosphoglucomutase activity by competing with magnesium, a cofactor of the enzyme. High magnesium concentrations revert lithium inhibition of growth and phosphoglucomutase activity. Lithium stress causes an increase of the phosphoglucomutase activity due to an induction of transcription of the PGM2 gene, and its overexpression confers lithium tolerance in galactose medium. These results show that phosphoglucomutase is an important in vivo lithium target.

Structure of an acidic exopolysaccharide produced by the diazotrophic endophytic bacterium Burkholderia brasiliensis.

The structure of an exopolysaccharide (EPS) produced by Burkholderia brasiliensis, a diazotrophic endophytic organism originally isolated from rice roots, has been determined. The bacterium was grown in a synthetic medium, containing mannitol and glutamate, which favours the expression of two anionic EPSs, which were separated by anion-exchange chromatography. The structure of the repeat unit of EPS A, eluted at higher ionic strength, was determined by a combination of methylation analysis, partial hydrolysis, chemical degradations, and NMR spectroscopic studies, and shown to be the linear O-acetylated pentasaccharide: -->4)-alpha-D-Glcp-(1-->2)-alpha-L-Rhap-(1-->4)-alpha-D-GlcpA-(1-->3)-beta-L-Rhap[2OAc]-(1-->4)-beta-D-Glcp-(1-->.

Autoantibodies specificity in acute anterior uveitis according to the presence of the HLA-B27 allele.

PURPOSE: To study HLA-B27(+) and HLA-B27(-) patients with AAU with respect to the frequency and specificity of organ-specific and non-organ-specific autoantibodies. METHODS: Fifty-seven consecutive patients with AAU were subjected to ophthalmologic and rheumatologic evaluation as well as to autoantibody determination: antinuclear antibodies (ANA), rheumatoid factor (RF), anticardiolipin, anti-smooth muscle, and anti-parietal cell antibodies, and immunoblot for antibodies to HeLa cells and to bovine iris extract. HLA-B27 was determined by a microlymphocytotoxicity assay. Statistical analysis employed chi-square test, Fisher's exact test, and McNemar test. RESULTS: Thirty-four patients had the HLA-B27 allele (B27(+)/AAU) and 23 did not (B27(-)/AAU). ANA, RF, anticardiolipin, and anti-parietal cell antibodies appeared in low frequency. No patient presented anti-smooth muscle antibody. Immunoblot showed a high frequency of antibodies to HeLa cell proteins in B27(-)/AAU patients with predominant reactivity at 46 kDa and 56 kDa. In contrast, sera from B27(+)/AAU patients reacted poorly against HeLa cell antigens. Immunoblot with bovine iris extract showed a significant frequency of antibodies in both groups, with a predominant response to antigens with an estimated mobility of 35, 52, and 54 kDa. CONCLUSION: Antibodies specific to iris antigens were equally frequent in both acute uveitis groups, whereas non-organ-specific autoantibodies, especially those to HeLa cell proteins, were far less frequent in B27(+)/AAU than in B27(-)/AAU patients.

Low-intensity exercise training decreases cardiac output and hypertension in spontaneously hypertensive rats.

The decrease in cardiac sympathetic tone and heart rate after low-intensity exercise training may have hemodynamic consequences in spontaneously hypertensive rats (SHR). The effects of exercise training of low and high intensity on resting blood pressure, cardiac output, and total peripheral resistance were studied in sedentary (n = 17), low- (n = 17), and high-intensity exercise-trained (n = 17) SHR. Exercise training was performed on a treadmill for 60 min, 5 times per week for 18 weeks, at 55% or 85% maximum oxygen uptake. Blood pressure was evaluated by a cannula inserted into the carotid artery, and cardiac output was evaluated by a microprobe placed around the ascending aorta. Low-intensity exercise-trained rats had a significantly lower mean blood pressure than sedentary and high-intensity exercise-trained rats (160 +/- 4 vs. 175 +/- 3 and 173 +/- 2 mmHg, respectively). Cardiac index (20 +/- 1 vs. 24 +/- 1 and 24 +/- 1 ml.min-1 x 100 g-1, respectively) and heart rate (332 +/- 6 vs. 372 +/- 14 and 345 +/- 9 beats/min, respectively) were significantly lower in low-intensity exercise-trained rats than in sedentary and high-intensity exercise-trained rats. No significant difference was observed in stroke volume index and total peripheral resistance index in all groups studied. In conclusion, low-intensity, but not high-intensity, exercise training decreases heart rate and cardiac output and, consequently, attenuates hypertension in SHR.

Pulmonary involvement in patients with HTLV-I associated myelopathy.

Pulmonary involvement in HTLV-I patients has been identified by several authors in Africa, Japan and Brazil. The objective of this controlled study is to establish an association between HAM and lymphocytosis in the bronchoalveolar fluid (BAL). Thirty-five adult patients with non-traumatic and non-tumoral myelopathy filled out a detailed historical survey and underwent a neurological examination, a thoracic radiological evaluation and a CSF examination. Of the patients in this sample, 22 were diagnosed with HTLV-I associated myelopathy and 13 had myelopathies with other etiologies. Lymphocytosis in the BAL fluid was detected in 18 (82%) of the HAM patients and in 2 (15%) of non-HAM patients. We concluded that the lung represents an important organ in the pathogenesis of HAM.

[HTLV-1 associated myelopathies in the city of Salvador, Bahia]. / Mielopatias por HTLV-1 na cidade de Salvador, Bahia.

Chronic myelopathy associated with human T-lymphotropic virus type I (HTLV-I) has been described in several endemic areas in Brazil. In Salvador, 102 patients with myelopathy were screened for HTLV-I/II by ELISA and Western blot assays. We found 36 patients with HTLV-I/II associated myelopathy confirming the high prevalence of HAM in Salvador. The initial complaint of our patients were urinary urgency, back pain and progressive unsteadiness on walking. On examination all of them had a spastic paraparesis, variable degrees of lower motor neuron syndrome, deep and superficial sensitive syndrome. MRI analysis revealed lesions in the periventricular white matter in addition to atrophy of the thoracic spinal cord. Clinical and magnetic resonance findings reveal that the inflammatory lesions of HAM involve not only the spinal cord but also the brain and peripheral nervous system.

PIP: During November 1990 and June 1992 a total of 102 patients with myelopathy who attended four hospitals in the city of Salvador, Bahia, Brazil, were studied. The human T-cell lymphotropic virus types I and II (HTLV-I/II) was obtained by means of enzyme-linked immonosorbent assay (ELISA) and Western blot tests. There were 36 patients who had HTLV-I/II associated myelopathy (HAM) of whom 26 were women (72%). The age range was 8 to 82 years, and the average age of those with HTLV-I myelopathy was 45.8 years. Previous history of blood transfusion was established in 6 patients. Only 3 patients had a history of promiscuity, defined as more than 5 partners in a year. The most frequent clinical symptoms included progressive weakness in the lower extremities (29/36), followed by lumbar pain (18/36), and urinary urgency (15/36). The progression of myelopathy was slow and gradual in 25 cases and more rapid in 11 cases, reaching severe muscular force in about 2 years. Except for 2 patients all others reported asymmetry in the muscular force of the lower extremities. On examination all of them had a spastic paraparesis, variable degrees of lower motor neuron syndrome, and deep and superficial sensitive syndrome. Clinical and magnetic resonance findings revealed that the inflammatory lesions of HAM involved not only the spinal cord but also the brain and the peripheral nervous system. Magnetic resonance examination of the thoracic column of 19 patients demonstrated encephalitic disorder in 11 patients. The cerebrospinal fluid (LCR) indicated cellular augmentation in 24 patients with the range of cells from 1 to 40 per cubic millimeter and with the predomination of lymphocytes. Electrophoresis of proteins of LCR showed the increase of albumin in 8 patients and the increase of gamma globulin in 29 patients. HIV antibodies were detected in 2 patients.

[HTLV-I-associated myelopathy: a systemic disease]. / Mielopatia por HTLV-I: uma doença sistêmica.

Chronic myelopathy associated with T-lymphotropic virus type I (HAM) has been described as an endemic disease in several areas of the world mainly in Asia, Africa and Latin America. The involvement of the central and the peripheral nervous system by HTLV-I observed in South America and Africa points out that this disease is a leukoencephalomyeloneuropathy. Several authors reported involvement outside the nervous system (as uveitis, lymphocytic alveolitis and arthritis, among others) in patients with HAM. In this paper we emphasize that HAM is a systemic disease whose cofactors determine the clinical picture in each patient.

Lymphocyte alveolitis in HAM/TSP patients: preliminary report

Mielopatia por HTLV-I (HAM) tem sido descrita como doença sistêmica caracterizada pelo acometimento de vários órgäos além do sistema nervoso. Neste registro, estamos relatando o envolvimento pulmonar em pacientes com HAM

Lymphocyte alveolitis in HAM/TSP patients. Preliminary report.

HTLV-I associated myelopathy has been described as a systemic disease characterized by manifestations in several organs outside the nervous system. We report inflammatory pulmonary involvement in patients with diagnosis of HAM.