Decatropis bicolor (Zucc.) Radlk essential oil induces apoptosis of the MDA-MB-231 breast cancer cell line.

BACKGROUND: Decatropis bicolor (Zucc.)Radlk is a plant that has been traditionally used for the treatment of breast cancer in some communities of Mexico. So, the aim of this study was to determine the cytotoxic and apoptotic effect of the essential oil of Decatropis bicolor against breast cancer cell line, MDA-MB-231. METHODS: The essential oil obtained from hydrodestillation of leaves of Decatropis bicolor was studied for its biological activity against breast cancer cells MDA-MB-231 by MTT assay, Hematoxylin-eosin stain, Annexin V-FITC, TUNEL and western blot assays and for its chemical composition by GC-MS. RESULTS: The results showed a relevant cytotoxic effect of the essential oil towards MDA-MB-231 cells in a dose- and time- dependent manner, with an IC50 of 53.81 ± 1.691 µg/ml but not in the epithelial mammary cell line MCF10A (207.51 ± 3.26 µg/ml). Morphological examination displayed apoptotic characteristics in the treated cells like cell size reduction, membrane blebbing and apoptotic bodies. In addition, the apoptotic rate significantly increased as well as DNA fragmentation and western blot analysis revealed that the essential oil induced apoptosis in the MDA-MB-231 cells via intrinsic pathways due to the activation of Bax, caspases 9 and 3. Phytochemical analysis of the Decatropis bicolor essential oil showed the presence of twenty-three compounds. Major components of the oil were 1,5-cyclooctadiene,3-(methyl-2)propenyl (18.38 %), ß-terpineol (8.16 %) and 1-(3-methyl-cyclopent-2-enyl)-cyclohexene (6.12 %). CONCLUSIONS: This study suggests that essential oil of Decatropis bicolor has a potential cytotoxic and antitumoral effect against breast cancer cells, with the presence of potential bioactive compounds. Our results contribute to the validation of the anticancer activity of the plant in Mexican traditional medicine.

Profilin sensitisation in a Mediterranean population.

BACKGROUND: Sensitisation to pan-allergens has become an interesting tool for the study of the allergenic profile of different populations. Profilins are one of the most common pan-allergens to be studied because they are responsible for a large number of sensitisations and are clearly related to cross-reactivity and co-sensitisation. OBJECTIVES: The objective of this study was to investigate the profile of sensitisation to profilins and to correlate it with sensitisation to foods and pollens. METHODS: Six hundred and fifty-four consecutive patients were skin-prick tested with a battery of common allergens including pollens, epithelia, mites and moulds and profilin and divided into three groups depending on their sensitisation profile (non-atopic, atopic with pollinosis and atopic without pollinosis). Patients with symptoms were challenged and diagnosed with the offending food extracts. Profilin sensitisation was identified and analysed in detail. RESULTS: According to the classification of the population, the prevalence of profilin sensitisation was estimated at 2.9% in patients suffering respiratory allergy, 4.2% in atopic patients, and 5.9% in pollen-sensitised individuals. Positive association was observed between pollen (except Cupressus and olive) and profilin but not with moulds, mites or epithelia. With respect to foods, positive association was only observed between profilin and melon sensitisation. Lastly, in terms of symptoms, positive association was only observed between profilin sensitisation and OAS. CONCLUSION: Profilin sensitisation seems to be a marker of pollen-related poly-sensitisation in our area. Pan-allergen diagnosis seems to be an essential tool for developing and improving selection of the correct treatment for allergic patients.

Differential expression of CD30 on CD3 T lymphocytes in patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune systemic disease caused as a result of an imbalance of Th1-/Th2-type cytokines. The soluble form of CD30 (CD30s) released from peripheral blood cells has been described as a marker of active disease in Th2-type immune response as in SLE. However, the expression of CD30 on CD3 T lymphocytes from patients with SLE has not been studied yet. Therefore, we have addressed our study to attempt this issue, studying CD30 expression by flow cytometry on CD3 T lymphocytes and CD4/CD8 subsets in samples from SLE patients mainly with lupus nephritis. In parallel, we have determined the production of the cytokines IL-4 (Th2), IFNγ (Th1), IL-10 and TGFß by intracellular staining. Differences between positive CD30 T cells in healthy controls and patients with SLE were found, with a higher percentage of CD30-expressing T cells in patients with SLE (P = 0.001). In contrast to healthy controls, CD30 was mainly expressed on CD8 T cells from patients with SLE. The intracellular cytokine staining showed that TGFß is the main cytokine expressed in CD3 T cells from patients with SLE. In addition to this, we have found a positive correlation between CD30-expressing T cells and IL-4, IFNγ, and immunosuppressive cytokines (IL-10 and TGFß) (P < 0.05). These results suggest that CD30 could play a role in the pathogenesis of SLE and its expression on CD3 T lymphocytes is not restricted only to Th2-type response.

Mycophenolate as induction therapy in lupus nephritis with renal function impairment.

BACKGROUND: Mycophenolate (MF) is effective as induction therapy for lupus nephritis (LN) in patients with normal renal function; however, little is known about its role in patients with impaired renal failure. The purpose of this study was to evaluate the response to MF in LN and its association with baseline renal function. METHODS: Data were obtained for 90 patients from 12 Spanish renal units who were receiving MF as induction therapy for LN. Patients were classified into 2 groups: group 1 (estimated glomerular filtration rate [eGFR] ≥60 ml/min/1.73 m(2)) and group 2 (eGFR <60 ml/min/ 1.73 m(2)). The primary outcome measure was the percentage of patients who achieved any response and its relationship with initial eGFR. The secondary outcome measures were the percentage of patients who achieved a complete response (CR) or partial response (PR) and the appearance of relapses during treatment and side effects. RESULTS: At initiation of MF treatment, there were no differences in the main parameters between group 1 (n = 63; eGFR 87 ± 23 ml/min/ 1.73 m(2)) and group 2 (n = 27; eGFR 44 ± 12 ml/min/1.73 m(2)). Exposure to prednisone and MF was similar. The percentages of patients who achieved a response in groups 1 and 2 were, respectively, 69.2 and 43.8% at 6 months and 81.3 and 73.7% at 12 months. CR was more frequent in group 1, whereas PR was similar in both groups. Four patients relapsed and side effects were unremarkable. CONCLUSIONS: MF is effective and safe as induction therapy for LN, and response is even achieved in patients with baseline renal impairment.

[Comorbidity, anemia and response to erythropoiesis stimulating agents in chronic hemodialysis]. / Relación entre comorbilidad, anemia y respuesta a derivados eritropoyéticos en pacientes incluidos en programa de hemodiálisis periódicas.

INTRODUCTION: Patients treated with haemodialysis have a high prevalence of co-morbidity that induces a elevate mortality risk. On the other hand, these patients have anaemia whose treatment is based in erythropoiesis stimulating agents. To date there are not enough studies to determine if co-morbidity alters erythropoietin response and the relationship between co-morbidity, response to treatment of anaemia and resistance to erythropoiesis-stimulating agents. OBJECTIVES: We have the following objectives: i) to study the prevalence of associated diseases in patients treated with haemodialysis in our Hospital Unit and to evaluate the co-morbidity Charlson Index, ii) to know the degree of anaemia control, dose and response to erythropoiesis-stimulating agents, and iii) to determine the relationship with co-morbidity and anaemia treatment. PATIENTS AND METHODS: We designed a retrospective study in stable haemodialysis treated patients. We calculated the Charlson co-morbidity index adjusted to age and we analysed levels of haemoglobin in the 6 months before study, dose of erythropoiesis-stimulating agents and its resistance index defined as doses of erythropoiesis-stimulating agents/weight (kg)/week/haemoglobin (g/dL). The different variables included in Charlson index were considered as independent variables and the index to repose to erythropoiesis-stimulating agents as a dependent variable, using bivariant and multivariate statistical analysis. RESULTS: We included 58 patients (31 males and 27 females), median age of 69.5 years (range 24-88), mean haemodialysis 83.7 months. Mean Charlson index was 7.4 +/- 2.8 (range 2-13). Comorbidity-age Charlson index was 2 in 3.4% of patients; 10.3% had 3 or 4 points; 43.2% between 5 and 7 and 43,1% 8 or more. Mean haemoglobin levels was 11,7+/-1,2 g/dL. Mean erythropoiesis-stimulating agents dose was 163.7+/-114.5 IU/kg/week and resistance index 14.1+/-9.7. Most of patients (57%) had a IRE value higher than 10. Fourteen patients (24%) had haemoglobin less than 11 g/dL, and 3 of them (5.1%) received erythropoiesis-stimulating agents more than 300 IU/kg/week. Nine subjects (15.5%) was treated with high dose of erythropoiesis-stimulating agents (>300 IU/kg/week): 3 of them had Hb>or=11 g/dL and 6 had Hb<11 g/dL. We did not found that the intensity of Charlson index is related with the degree of anaemia control or response to erythropoiesis-stimulating agents. CONCLUSIONS: Although the co-morbidity index is high and the response to erythropoiesis-stimulating agents is inadequate. In our study there is not relationship between these conditions.

Inhibitory effect of biofilm-forming Lactobacillus kunkeei strains against virulent Pseudomonas aeruginosa in vitro and in honeycomb moth (Galleria mellonella) infection model.

Biofilms correspond to complex communities of microorganisms embedded in an extracellular polymeric matrix. Biofilm lifestyle predominates in Pseudomonas aeruginosa, an opportunistic Gram negative pathogen responsible for a wide spectrum of infections in humans, plants and animals. In this context, anti-biofilm can be considered a key strategy to control P. aeruginosa infections, thereby more research in the field is required. On the other hand, Lactobacillus species have been described as beneficial due to their anti-biofilm properties and their consequent effect against a wide spectrum of pathogens. In fact, biofilm-forming Lactobacilli seem to be more efficient than their planktonic counterpart to antagonise pathogenic bacteria. In this work, we demonstrated that Lactobacillus kunkeei, a novel Lactobacillus species isolated from honeybee guts, can form biofilms in vitro. In addition, the L. kunkeei biofilm can, in turn, inhibit the formation of P. aeruginosa biofilms. Finally, we found that L. kunkeei strains attenuate infection of P. aeruginosa in the Galleria mellonella model, presumably by affecting P. aeruginosa biofilm formation and/or their stability. Since L. kunkeei presents characteristics of a probiotic, this work provides evidence arguing that the use of this Lactobacillus species in both animals (including insects) and humans could contribute to impair P. aeruginosa biofilm formation.

[Factors which influence phosphorus removal in hemodialysis]. / Factores en la eliminación de fósforo en hemodialisis.

BACKGROUND: The sustained elevation of phosphorous among patients with end-stage renal failure is associated with elevated mortality rates. Phosphate binding agents are usually necessary to control serum phosphate levels. Phosphate removal during dialysis is limited largely due to the intracellular location of most inorganic phosphorous. The membrane surface, the frequency and the duration of therapy have proved to be very important factors in the serum phosphate control. THE AIM of our work is to investigate the influence on phosphate removal of factors that normally participate in the haemodialysis session: Plasma phosphate level (Php), treatment duration, membrane surface, high or low-flux membranes, the vascular access, dialysate flux , the volume of blood passing through the dialyzer (L) in each dialysis session and the blood flow during the first hour of dialysis. On 16 patients, we also had the possibility of comparing phosphate removal with 1.8 m(2) high-flux haemodialysis, 1.8 m(2) on-line hemodiafiltration and the on-line technique with the new Helixone dialyzer Fresenius Fx100. METHODS: 108 haemodialysis patients, 62% men, 38% women aged 21-82 years (61+/-14;mean+/-sem),) were selected for the study. Mean treatment time 4.14+/-0.41 hours (range 3.5-5 hours). The vascular access was an arterio-venous fistula in eighty five (78%) and a double lumen tunnelled catheter 23 (22%). Patients were studied under their normal every day conditions. High-flux membrane was used by 31 (30%) patients and low-flux membrane by 77 (70%). Membrane surface was: 1.7 m2:17 (16%); 1.8 m2:77 (71%); 2,1 m2:14 (13%). Dialysate flux was: 500 ml/min. 55 patients; 700 ml/min: 53 patients. In 16 out of 108 patients we had the possibility of using on-line hemodiafiltration with ultrapure bicarbonate-buffered dialysate. Phosphate mass removal (MPO4) was calculated using the formula:MPO4=0.1 t-17+50 Cds 60+11Cb 60 (1), where t is treatment time in minutes, Cds60 and Cb60 are phosphate concentrations in dialysate and plasma measured at 60 min from the beginning of hemodialysis in mg/dl, and MPO4 is the estimated phosphate removed in mg/treatment. RESULTS: We found a good correlation between phosphate removal and serum phosphate levels (p=0.01), but not with the membrane surface or treatment duration. Phosphate removal was 640+/-180 mg/session with low-flux membrane and 700+/-170 mg/session with high-flux membrane (p=0.280). The MPO4 was 720+/-190 mg/treatment in patients with a AV fistula and 620+/-180 in patients with a tunnelled catheter (p=0.023). We found a good correlation between phosphate removal and the volume of blood (L) that passed the dialyzer in each session (r=0.001) but we did not find a correlation between phosphate removal and KT/Vurea, the dialysate flux or the ultra filtration. On-line technique did not increased the MPO4(733+/-280 mg, p=0.383). The on-line technique with the new dialyzer (Fresenius Fx100), increased the phosphate removal to 759+/-199 mg/session (p=0.057). CONCLUSION: Phosphate removal during dialysis is influenced by Plasma phosphate levels, the volume of blood that passed the dialyzer and the vascular access. Uniformity on time and membrane surface could explain the abs cense of influence in our case. The ultra filtration, dialysate flux, membrane permeability or on-line hemodiafiltration does not influence the phosphate removal. The new membrane helixone with 2,1 m2 (Fresenius Fx100) increases phosphate removal probably because the membrane surface is higher.

Five cases of acrometastasis to the hand from a carcinoma and review of the literature.

Metastases in the hand bones are a rare form of cancer presentation. Their appearance as a sign of carcinoma is even rarer and is associated with a poor prognosis. While amputation is recommended in cases of isolated metastases in patients with at least a few months of survival, radiation therapy may be useful for treating pain and partially restoring function. We conducted a retrospective review of 5 consecutive patients (2 male, 3 female; mean age of 46 years) presenting with metastases in the hand bones who had lung (n=2), skin, uterus and kidney cancers. Conservative treatment was performed in three cases, transmetacarpal amputation in one case and distal phalanx amputation in one case. All patients died within a few months of the diagnosis (mean: 5.2months). Because acrometastases generally are related to widespread disease, the prognosis of patients with acrometastases is poor. These cases illustrate the rapid progression of the disease when acrometastases in the hand are present.

High throughput sequencing and comparative genomics of foot-and-mouth disease virus.

Despite a basic understanding of many aspects of FMD biology, much information regarding FMDV virulence, host range, and virus transmission remains poorly understood. Here we present how the use of high throughput sequencing for complete genome sequences of foot-and mouth disease virus (FMDV) led to a series of new insights into viral genome sequence conservation and variability, genetic diversity in nature and phylogenetic classification of isolates, including the first complete sequences of the South African Territories type 1 and 3 (SAT1 and SAT3) genomes. Comparative genomic analysis of full-length sequences of FMDV isolates did allow: (i) the identification of highly conserved regulatory or coding regions which are critical for aspects of virus biology as well as novel viral genomic motifs with likely biological relevance; (ii) characterization of the first complete sequences of the SAT1 and SAT3 genomes; (iii) identification of a novel SAT virus lineage genetically distinct from other SAT and Euro-Asiatic lineages; (iv) precise identification of strains circulating around the world for epidemiological and forensic attribution; (v) assessment of mutation and recombination processes as mechanisms equally involved in evolution; (vi) mutation rates, tolerance and constraints of genes and proteins during evolution of FMD viruses during in vivo replication and (vi) support for the hypothesis of a new evolutionary model.

[CA-P control in haemodialysis and K/DOQI guidelines]. / Control del metabolismo calcio-fósforo en hemodiálisis y su adecuación a las guías K/DOQI 2003.

BACKGROUND: The publication in 2003 of the K/DOQI Clinical Practice Guidelines for Bone Metabolism and Disease in Chronic Kidney Disease recommended targets levels for serum iPTH, Ca, P, and CaxP product. However, many patients do not achieved these target ranges. It is necessary to known the percentage of patients out of range in order to prevent the development of bone disease and to reduce mortality and morbidity. OBJECTIVES: To know the degree of control of Ca-P metabolism in haemodialysis patients in our haemodilalysis facilities and the achievement of target levels recommended by K/DOQI Guidelines. PATIENTS AND METHODS: We have retrospectively investigated in 190 prevalent haemodialysis patients (males 58.2%, ratio M/F 1.4, mean age 70 years, range 17-87 years, at least 3 months in haemodialysis) the serum levels of Ca, albumin-corrected serum Ca, P, CaxP product and iPTH in all analitycal determinations performed in 2004. In each patient we have obtained the average (and median) of these serum markers. Cut-off levels were carried out following the recommendations of the K/DOQI Guidelines. RESULTS: The average of serum Ca and albumin-corrected serum Ca is normal (means +/- SD = 8.9 +/- 0.6 mg/dL and 9.2 +/- 0.7 mg/dL, respectively); however, 53.7% has normal values, 9.1% hypocalcemia and 37.1% hypercalcemia. The average of serum P is also normal (mean +/- SD = 5.0 +/- 1.3 mg/dL); however, only 57.2% has normal values, and 11.7% has hypophosphoremia and the remaining 31, 1% hyperphosphoremia. The CaxP product is normal (mean +/- SD = 46.3 +/- 13.3 mg2/mL2), 4.9% with low values and 23.4% with high values. The median of serum iPTH is 253 pg/mL, but only 31.1% of them have normal values, 25.1% low range values and 43.7% has hyperparathyroidism; 9.3% with iPTH higher than 800 pg/mL. The percentage of patients with hyperphosphoremia is higher in the group with iPTH higher than 300 pg/mL (23.3% vs. 40%, chi2, p= 0.006). In patients with PTHi in normal range, 3.6% have low CaxP product and the remaining 17.8% high CaxP product. Overall, only 25% of patients falls within recommended ranges for all indicators of mineral metabolism and 17% has all serum markers outside these recommendations. CONCLUSIONS: The degree of control of mineral metabolism in haemodyalisis patients if clearly insufficient and a large percentage of them do not achieved the recommended serum targets recommended by K/DOQI Guidelines. This groups of patients are exposed to a increased risk for oseous and cardiovascular morbimortality. The analysis of adequacy must be performed with percentage of patients out of range in order to apply new therapeutical strategies.

Reperfusion arrhythmias after chronic regional denervation of the ischaemic myocardium in pigs.

OBJECTIVE: The aim was to assess the effects of chronic regional denervation of the ischaemic myocardium on reperfusion arrhythmias in a model with sparse coronary collateral circulation. METHODS: Baseline ventricular refractoriness and epicardial activation times were measured together with reperfusion arrhythmias after 15 min (I-15') or 30 min (I-30') of left anterior descending coronary artery occlusion in 38 barbiturate anaesthetised open chest pigs. Twenty pigs (11 in I-15' and nine in I-30') had a chronic (two week) denervation of the left anteroseptal region, whereas 18 pigs (10 in I-15' and eight in I-30') were sham operated (non-denervated) controls. Denervation was induced by pericoronary application of phenol and verified by absence of adrenergic histofluorescence. RESULTS: As compared with controls, denervated pigs showed: (1) longer activation times: 20.3 (SD 5.2) ms v 16.5 (4.6) ms, p < 0.001; (2) slightly longer refractory periods: 348(28) ms v 334(27) ms; (3) a tendency to lower postreperfusion ectopic activity: ectopic beats divided by time free of ventricular tachycardia: 0.13(0.19) v 0.34(0.40) in I-15', and 0.21(0.24) v 0.39(0.44) in I-30'; (4) slower ventricular tachycardia in I-30': 140(29) beats.min-1 v 185(29) beats.min-1, p < 0.009; and (5) comparable incidence of postreperfusion ventricular fibrillation: 4/11 pigs v 2/10 in I-15', and 5/9 v 4/8 in I-30'. CONCLUSIONS: Selective chronic denervation of the ischaemic myocardium was unable to protect against malignant reperfusion arrhythmias in hearts with human-like coronary collaterals. This was confirmed at two ischaemic periods known to produce progressive catecholamine accumulation and increased adrenoceptor density in the ischaemic myocardium.

Coronary denervation attenuates coronary constriction induced by muscarinic receptor stimulation in pigs.

OBJECTIVES: We tested the hypothesis that coronary denervation attenuates the reactivity of the coronary vessel to cholinergic stimulation. METHODS: Heart rate, left ventricular (LV) pressure, LV dP/dt, coronary blood flow at the left anterior descending (LAD) coronary artery, and epicardial ECG mapping were measured before and after topical application of 1% methacholine to the LAD in 10 pigs anesthetized with alpha-chloralose (100 mg/kg, i.v.); these were compared with 10 other pigs submitted 2 weeks previously to a denervation of the LAD with phenol. Coronary denervation was confirmed in all cases by adrenergic histofluorescence and by acetyl-cholinesterase staining. Isolated LAD rings from 10 additional pigs (5 controls and 5 treated with phenol) were stimulated with endothelin-1 to verify whether phenol affected coronary reactivity to noncholinergic stimulation. RESULTS: Methacholine induced a fall in coronary blood flow (10.3 +/- 5.3 ml/min vs 4.8 +/- 6.2 ml/min, ANOVA: P < 0.001), a drop in systolic LV pressure (113 +/- 19 mmHg vs 93 +/- 19 mmHg, P < 0.001) and LV dP/dt (1608 +/- 363 mmHg/s vs 1203 +/- 302 mmHg/s, P = 0.02) and elevation of the ST segment (1.4 +/- 0.9 vs 11.1 +/- 4.7 mV, P < 0.001) in controls. These changes were not preceded by heart rate variations and were inhibited by atropine. As compared to controls, phenol-treated pigs showed a smaller decline in coronary blood flow (13.1 +/- 4.5 ml/min to 10.4 +/- 5.4 ml/min, P < 0.001), a lower drop in LV pressure (107 +/- 20 mmHg to 100 +/- 19.7 mmHg, P < 0.001) and lesser ST segment elevation (2.2 +/- 1.7 mV to 5.6 +/- 4.2 mV, P < 0.001). Isolated LAD rings contracted after exposure to endothelin-1 in both controls and phenol-treated pigs (3.5 +/- 0.7 g vs 2.4 +/- 1.0 g, P = 0.06). CONCLUSIONS: Coronary denervation attenuates coronary constriction induced selectively by direct muscarinic receptor stimulation in the in situ pig heart.

Passive transmission of ischemic ST segment changes in low electrical resistance myocardial infarct scar in the pig.

OBJECTIVES: To analyze the passive electrical properties of a healed infarction and assess their role on transmission of contiguous ischemic ST segment potential changes. METHODS: We measured tissue resistivity (omega cm) at 1 kHz and the epicardial ST segment during 1 h of proximal reocclusion of the left anterior descending (LAD) coronary artery in 12 anesthetized pigs with one-month-old transmural infarction elicited by LAD ligature below the first branch. The impedance spectrum (1 to 1000 kHz) of normal and infarcted myocardium was measured in seven other pigs with similar infarctions. Electrical transmission of current pulses (30 microA) in infarcted tissue and in test solutions was also investigated. RESULTS: The infarct scar has a lower than normal resistivity (110 +/- 30 omega cm vs. 235 +/- 60 omega cm, p < 0.0001) and, unlike the normal myocardium, resistivity and phase angle of the scar did not change at increasing current frequencies, reflecting no capacitative response. LAD reocclusion induced a resistivity rise (510 +/- 135 omega cm, p < 0.01) and a ST segment elevation (0.6 +/- 0.7 to 9.5 +/- 5.1 mV, p = 0.002) in the ischemic peri-infarction zone, whereas the infarcted area showed ST segment elevation (0.5 +/- 0.5 to 3.8 +/- 2.6 mV, p = 0.03) with no resistivity changes. Potential decay of both ST segment and current pulses in the scar and in 0.9% NaCl solution was less than 1 mV/mm. Transmural deposition of connective tissue was seen in the center of the infarction. CONCLUSIONS: A one-month-old transmural infarction is a low resistance, noncapacitative medium that allows a good transmission of current pulses and of ST segment potential changes generated by contiguous peri-infarction ischemia.

Low incidence of ventricular arrhythmias induced by ischaemia at the borders of a chronic infarct in a model with local postinfarction denervation.

OBJECTIVE: The aim was to assess the arrhythmogenic potential of acute ischaemia superimposed at the borders of a chronic myocardial infarct and to analyse the effects of myocardial necrosis on local autonomic innervation in pigs. METHODS: Ventricular arrhythmias were measured in alpha chloralose (100 mg.kg-1) anaesthetised open chest pigs during 60 min occlusion of the left anterior descending coronary artery 2 cm above the first diagonal branch (group I, n = 11) or just below this branch (group II, n = 12). These arrhythmias were compared with those induced in pigs with a one month old anteroseptal infarction (coronary ligature as in group II) submitted to a second occlusion 2 cm above the first (group III, n = 12). The area at risk after high or low ligature was measured in 12 control pigs using fluorescein. Sympathetic and parasympathetic innervation of the anteroseptal myocardium was studied in three pigs with a chronic anteroseptal infarction and in six pigs without infarction using adrenergic histofluorescence and acetylcholinesterase reaction. RESULTS: Compared with ischaemia alone, ischaemia at the borders of a chronic infarct induced a lower incidence of ventricular fibrillation (1/12 pigs v 11/11 in group I, p < 0.001, or 6/12 in group II, p < 0.05) and a tendency towards a lower occurrence of ventricular tachycardia (2/12 pigs v 8/11 in group I, p = 0.01, and 4/12 in group II) and fewer ventricular premature beats (mean number: 105 in group I v 30 in group III, p < 0.05). The mass of the ischaemic regions after low or high occlusion was 13.3(SD 3.0) g and 23.2(5.8) g, respectively. Adrenergic and cholinergic denervation was observed inside the necrotic area, along the subendocardium surviving the necrosis, and in a band of normal bordering myocardium [width: 3.2(2.0) mm for adrenergic and 2.1(1.2) mm for cholinergic denervation]. CONCLUSIONS: Acute ischaemia at the borders of a chronic anteroseptal infarct has a low arrhythmogenic potential in pigs. In this model the peri-infarction zone shows a band of sympathetic and parasympathetic denervation secondary to the necrosis.

Differential uptake of myocardial perfusion radiotracers in normal, infarcted, and acutely ischemic peri-infarction myocardium.

OBJECTIVES: We measured the uptake of technetium-99m tetrofosmin (99m Tc) and thallium-201 (201 TI) in areas of healed transmural myocardial infarction and in the regions of acute peri-infarction ischemia. METHODS: Anesthetised pigs with a 1-month old transmural infarction elicited by permanent ligature of the left anterior descending (LAD) coronary artery below the first branch underwent one hour of proximal LAD occlusion followed by injection of 99m Tc-tetrofosmin and 201TI either in the left atrium (GI, n= 19) or in the jugular vein (GII, n = 6). Twelve other pigs (GIII) with similar acute peri-infarction ischemia received 99m Tc-tetrofosmin and 201Tl into the left ventricle during cardiocirculatory arrest to rule out the effect of coronary collaterals. Radiotracer counting was determined in samples from normal, acute ischemic and necrotic regions. RESULTS: Uptake of 99m Tc-tetrofosmin and 201 Tl was greater in the infarct scar (median % of normal tissue: 20 for 99m Tc and 8.6 for 201 Tl in GI; 22 and 15 in GII) than in acute ischemic myocardium (3.2 and 2.5 in GI; 6.4 and 3.3 in GII). Radiotracer injection in arrested hearts (GIII) depicted a similar pattern (median % of injected dose: 6.2 for 99m Tc and 10 for 201Tl in the scar; 2.3 and 4.0 in acute ischemia; 2.9 and 3.5 in normal tissue). The infarcted region showed connective tissue and lack of viable myocardium. CONCLUSION: A 1-month old infarct scar with no viable myocardial tissue can take up significant fractions of 99mTc-tetrofosmin and 201Tl even in the absence of coronary collateral perfusion. Data suggest that the infarct scar can extract these radiotracers from the intraventricular blood.

Familial microscopic hematuria caused by hypercalciuria and hyperuricosuria.

We report 12 patients belonging to five different families in whom persistent isolated microhematuria was associated with hypercalciuria and/or hyperuricosuria. Four patients had episodes of gross hematuria, three patients had passed renal stones, and a history of nephrolithiasis was obtained in four of the families (80%). Calcium oxalate and uric acid crystals were commonly observed in the urine sediments. Urinary erythrocytes had a normal appearance on phase-microscopic examination. Reduction of calciuria and uricosuria by thiazide diuretics, allopurinol, forced fluid intake, and dietetic measures led to a persistent normalization of urine sediment with complete disappearance of hematuria. Determination of calcium and uric acid urinary excretions should be included in the study of familial hematuria.

Secondary (AA-type) amyloidosis in patients with polymyalgia rheumatica.

Several cases of systemic amyloidosis associated with polymyalgia rheumatica (PMR) or giant-cell arteritis (GCA) have been described. Nevertheless, the type of amyloid deposit has not been characterized in most of them. Here we report on two patients with PMR (one with associated GCA) who developed nephrotic syndrome and end-stage renal failure caused by massive amyloid deposition. Immunohistochemical analysis showed that the amyloid deposits were of AA type (secondary amyloidosis) in both cases.

In vivo and in situ ischemic tissue characterization using electrical impedance spectroscopy.

The investigation of processes of ischemia in different organ tissues is very important for the development of methods of protection and preservation during surgical procedures. Electrical impedance spectroscopy was used to distinguish between different tissues and their degree of ischemia. We describe mathematical methods used to adjust experimental data to Cole-Cole models for one-circle and two-circle impedance loci and a study of the main parameters for representing the behavior of ischemia in time. In vivo and in situ postmortem measurements of different tissues from pigs are shown in the 100 Hz to 1 MHz range. The Cole parameters that best characterize the ischemia are R0 and fc.

Intraoral mandibular distraction osteogenesis: special attention to treatment planning.

PURPOSE: To demonstrate our experience using internal devices for unidirectional distraction osteogenesis in treating different mandibular hypoplasias (with or without maxillary deformities). An algorithmic table for diagnosis, and treatment planning is presented. PATIENTS AND METHODS: Twenty internal distraction devices were used in 16 patients with mandibular hypoplasia. Deficiency in length of the mandible was calculated on three-dimensional computed tomography scans. The device was activated by a transcutaneous pin on the fifth postoperative day. Distraction was achieved at rates of 0.5 mm/12 h. After a variable period of consolidation the devices were removed. Mean follow-up was 18 months. RESULTS: Successful distraction osteogenesis was achieved in all patients. No premature consolidation or pseudoarthrosis was observed. Improvement of facial aesthetics was produced in all cases. Final occlusion was excellent in those cases where no simultaneous maxillary deformity was present. Orthodontic treatment was applied in all cases. Results remained stable one year postoperatively. CONCLUSIONS: The occlusal results obtained in this series show that we can plan distraction as a definitive treatment in cases with isolated mandibular hypoplasia. When an additional maxillary deformity is present, mandibular distraction must be performed first if indicated, but a maxillary procedure will be necessary later.

[Diagnostic and clinical course features of lupus membranous glomerulonephritis]. / Peculiaridades diagnósticas y evolutivas de la glomerulonefritis membranosa lúpica.

BACKGROUND: Lupus membranous glomerulonephritis (LMG) is sometimes difficult to distinguish from idiopathic membranous glomerulonephritis (IMG) and little is known about its natural history. METHODS: We have performed a retrospective study of 16 patients with LMG, types Va ("pure" LMG) (8 patients) and Vb (mesangial proliferation) (8 patients). Analytical and clinical characteristics were analyzed in each case at the onset of the disease and during follow-up. RESULTS: At the time of diagnosis, 7 patients (43%) did not present any extrarenal manifestations of systemic lupus erythematosus (SLE). Nephrotic syndrome was the most common clinical presentation (81%). 14 patients (87%) showed positive antinuclear antibodies (ANA), but at low titres (< 1/300); anti-DNA antibodies were positive only in 3 cases (18%) and hypocomplementemia was observed in 2 (12%). The most relevant histological findings were the presence of Clq deposits (83%) and subendothelial and mesangial deposits. Treatment consisted of corticosteroids, adding immunosuppression when no response was obtained in 2-3 months. In recent years, an angiotensin-converting enzyme inhibitor (ACEI) was added before immunosuppression. At the end of the study, 6 patients (42%) developed chronic renal insufficiency and 8 (58%) renainea with normal renal function. Actuarial renal survival was 100% at 5 years and 75% at 10 years. The evolution towards a non-nephrotic range proteinuria throughout the follow-up was the only parameter significantly associated with a good prognosis. CONCLUSIONS: LMG can present without extra-renal symptoms of SLE and even with very poor expression of its serological markers. Almost half of the patients (42%) developed chronic renal insufficiency. The evolution towards a non-nephrotic range proteinuria correlated with the maintenance of a normal renal function.