Correction of a Complex Foot Deformity With V and Y Osteotomy and Ilizarov Method.

Many authors consider the Ilizarov method as the best choice for correction of complex multiplanar foot deformities. Fifty-one patients, for a total of 55 feet, underwent V or Y osteotomy (respectively 32 and 23), 12 were due to congenital clubfoot outcomes, 11 were from post-traumatic pathologies, 9 from hemimelia, 7 from Charcot-Marie-Tooth, 5 from poliomyelitis, 3 from spina bifida, 2 from myopathy, 2 from poly-epiphyseal dysplasia, 1 from achondroplasia, 1 from arthrogryposis, 1 patient from Charcot's foot and 1 rheumatoid arthritis. All patients were contacted to undergo AOFAS and EQ-5D-5L questionnaire. The mean distraction time was 74.0 days (±25.3) and it was longer for V osteotomy (63.1 ± 21.1 vs 81.8 ± 25.4 days for Y and V respectively, p = .006). The average fixation time was 97.9 days (± 61.2), 90 days in the Y and 103 days in the V osteotomy groups. The mean preoperative foot length was longer in the group of Y osteotomy patients (158.7 ± 21.3 mm vs 133.5 ± 21.6 mm, p < .001). The mean length postoperatively was substantially the same (Y group 164.0 mm vs V group 167.4 mm, p < .562). The V osteotomy produces an elongation ratio of 440% with respect to that produced from the Y osteotomy (ratio length/distraction V/Y 0.44/0.1), with only 19% more in the distraction days. (81.8 V group vs 63.1 Y group). While the V osteotomy allows a lengthening of about 27%, the Y osteotomy, despite having correction potential superimposable to the V osteotomy, minimizes the elongation component.

Content of cardiolipin of the membrane and sensitivity to cationic surfactants in Pseudomonas putida.

AIMS: To establish the role of cardiolipin (CL) of the membrane in response to the presence of tetradecyltrimethylammonium in Pseudomonas putida A (ATCC 12633). METHODS AND RESULTS: Two ORFs of Ps. putida A (ATCC 12633), which in Ps. putida KT2440 encode the putative CL synthase genes cls and cls2, were cloned, sequenced and mutated. Only the double mutant lacking cls and cls2 showed a reduction of the CL content, 83% lower than the amount produced by the wild-type. Accompanying this change was a 40% decrease in the content of unsaturated fatty acid. Consequently, the membrane of the mutant was more rigid than the one of the parental strain, as observed using fluorescence polarization techniques. The mutant strain showed reduced viability in the presence of tetradecyltrimethylammonium. The incorporation of exogenous CL into its membrane relieved sensitivity to the cationic detergent. CONCLUSIONS: Pseudomonas Putida cells with low levels of CL die in the presence of tetradecyltrimethylammonium, because they cannot counter the fluidizing effect of the cationic surfactant. SIGNIFICANCE AND IMPACT OF THE STUDY: The modification in the membrane phospholipids composition allows knowing the adaptation strategy of Ps. putida when these bacteria are exposed to cationic surfactant.

Physiological role of phosphatidylcholine in the Pseudomonas putida A ATCC 12633 response to tetradecyltrimethylammonium bromide and aluminium.

AIMS: To evaluate the effect of tetradecyltrimethylammonium bromide (TTAB) and aluminium stresses on the phospholipid (PL) composition of Pseudomonas putida A ATCC 12633. METHODS AND RESULTS: Pseudomonas putida were grown with TTAB in the presence or absence of AlCl(3), and the PL composition was analysed. The presence of TTAB resulted in an increase in phosphatidylglycerol and phosphatidic acid levels (6- and 20-fold, respectively) with respect to the levels in cells grown without the surfactant. With AlCl(3), phosphatidylcholine (PC) increased (threefold) and cell-free extracts contained approximately threefold more phosphatidylcholine synthase activities than extracts without AlCl(3), indicating that the PC level is dependent upon activation of this enzyme. CONCLUSIONS: The negative charges of the headgroups of PL are the primary membrane-associated factors for the response to TTAB. PC are involved in cellular responses to binding Al(3+) and should be viewed as a temporary reservoir of available Al(3+) to allow a more efficient utilization of TTAB by Ps. putida. SIGNIFICANCE AND IMPACT OF THE STUDY: The changes in the PL of Ps. putida in the presence of TTAB and AlCl(3) indicate that different responses are utilized by bacteria to maintain optimal PL composition in the presence of such environmental pollutants.

A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C-N bond with the production of trimethylamine.

This article describes a simple fluorescence method for the determination of tetradecyltrimethylammonium mono-oxygenase (TTAB mono-oxygenase) activity involving N-dealkylation of tetradecyltrimethylammonium bromide with concomitant production of trimethylamine (TMA). Activity was determined by measuring the formation of TMA using the morin reagent and aluminum (Al). Morin reacts with Al to form a fluorescent complex, Al-morin. In the presence of TMA, Al is tightly associated with TMA and cannot be sequestered by morin, thus providing evidence for formation of the Al-TMA complex. The concentration of TMA is estimated by calibration graphs constructed by plotting the fluorescence intensity of the Al-morin complex versus TMA concentration. The fluorescence intensities of the Al-morin complexes quenched by TMA are linearly dependent on both the time of the TTAB mono-oxygenase reaction and the amount of protein used in the reaction. The kinetic behavior is characterized by K0.5=4.26x10(-4) M, and the apparent Hill coefficient (napp)=2.24. These values are both comparable to those determined by GC-MS (K0.5=4.41x10(-4) M and napp=2.35). The advantages of this assay include rapid and efficient implementation and potential employment for routine accurate determinations of TTAB mono-oxygenase activity over a wide range of substrate concentrations.

Degradation of tetradecyltrimethylammonium by Pseudomonas putida A ATCC 12633 restricted by accumulation of trimethylamine is alleviated by addition of Al 3+ ions.

AIMS: To establish if tetradecyltrimethylammonium (TDTMA) might be degraded by pure culture of Pseudomonas strains, and how the presence of a Lewis' acid in the medium influences its biodegradability. METHODS AND RESULTS: From different strains of Pseudomonas screened, only Pseudomonas putida A ATCC 12633 grows with 50 mg l(-1) of TDTMA as the sole carbon and nitrogen source. A monooxygenase activity catalyzed the initial step of the biodegradation. The trimethylamine (TMA) produced was used as nitrogen source or accumulated inside the cell. To decrease the intracellular TMA, the culture was divided, and 0.1 mmol l(-1) AlCl(3) added. In this way, the growth and TDTMA consumption increased. The internal concentration of TMA, determined using the fluorochrome Morin, decreased by the formation of Al(3+) : TMA complex. CONCLUSIONS: Pseudomonas putida utilized TDTMA as its sole carbon and nitrogen source. The TMA produced in the initial step of the biodegradation by a monooxygenase activity was used as nitrogen source or accumulated inside the cell, affecting the bacterial growth. This effect was alleviated by the addition of AlCl(3). SIGNIFICANCE AND IMPACT OF THE STUDY: The use of Lewis' acids to sequester intracellular amines offers an alternative to achieve an efficient utilization of TDTMA by Ps. putida.

Modification of phospholipid composition in Pseudomonas putida A ATCC 12633 induced by contact with tetradecyltrimethylammonium.

AIMS: The aim of this work was to establish if the response to tetradecyltrimethylammonium (TDTMA), a representative quaternary ammonium compound (QAC), involves changes in the phospholipid (PL) composition of Pseudomonas putida A ATCC 12633. METHODS AND RESULTS: Pseudomonas putida was exposed to 50 mg l(-1) of TDTMA for 15 min, and PL composition was analysed. With respect to control values, phosphatidic acid and phosphatidylglycerol increased by 140% and 120%, respectively; cardiolipin decreased about 60%. In TDTMA-adapted bacteria, the most significant change was a 380% increase in phosphatidic acid. Accompanying this change was a 130% increase in phosphatidylglycerol and a 70% decrease in cardiolipin. The changes in adapted cells were reverted after two subcultures without biocide. CONCLUSIONS: Pseudomonas putida responded to TDTMA through quantitative changes in PLs with specific variations in the content of phosphatidic acid, phosphatidylglycerol and cardiolipin. These modifications indicated that these PLs are involved in cellular responses to QACs, utilizing phosphatidic acid principally to neutralize the high positive charge density given for the ammonium quaternary moiety from TDTMA. SIGNIFICANCE AND IMPACT OF THE STUDY: The changes in PL composition give a new insight about the response inflicted by Ps. putida when these bacteria are exposed to QACs.

Changes in enzymatic activities involved in glucose metabolism by acyl-CoAs in Trypanosoma cruzi.

This study describes the effect of some saturated and unsaturated free fatty acids and acyl-CoA thioesters on Trypanosoma cruzi glucose 6-phosphate dehydrogenase and hexokinase activities. Glucose 6-phosphate dehydrogenase was sensitive to the destabilizing effect provoked by free fatty acids, while hexokinase remained unaltered. Glucose 6-phosphate dehydrogenase inhibition by free fatty acids was dependent on acid concentration and chain length. Both enzymes were inhibited when they were incubated with acyl-CoA thioesters. The acyl-CoA thioesters inhibited glucose 6-phosphate dehydrogenase at a lower concentration than the free fatty acids; the ligands glucose 6-phosphate and NADP+ afforded protection. The inhibition of hexokinase by acyl-CoAs was not reverted when the enzyme was incubated with ATP. The type of inhibition found with acyl-CoAs in relation to glucose 6-phosphate dehydrogenase and hexokinase suggests that this type inhibition may produce an in vivo modulation of these enzymatic activities.

Ventilatory and respiratory muscle responses to hypercapnia in patients with paraplegia.

To evaluate ventilatory and respiratory muscle responses to hypercapnia in patients with paraplegia with paralysis of abdominal muscles, we studied seven patients with complete transection of the midthoracic cord (Th6-Th7) and six normal subjects. Minute ventilation (V E) and mean inspiratory flow responses to hypercapnia were similar in normal subjects and patients with paraplegia, but in the latter, at any given level of end-tidal CO(2) partial pressure (PET(CO(2))), tidal volume (VT) was reduced and frequency was increased. In normal subjects during hypercapnia, end-expiratory transpulmonary pressure (PL) and abdominal volume at end expiration decreased markedly, whereas end-expiratory volume of the rib cage (Vrc,E) remained constant, suggesting progressive recruitment of abdominal muscles. In patients with paraplegia compared to normal subjects the decrease in end-expiratory PL was reduced, and it was associated with a decrease in Vrc,E, suggesting recruitment of rib cage expiratory muscles. For a PET(CO(2)) of 70 mm Hg the estimated expiratory muscle contribution to VT was 10.3 and 28.4% (p < 0.02) in patients with paraplegia and normal subjects, respectively. We conclude that the V E-CO(2) relationship is preserved in patients with paraplegia with the development of a rapid and shallow pattern of breathing. This suggests that expiratory muscle paralysis elicits adaptation of the ventilatory control system similar to that observed in patients with generalized respiratory muscle weakness.

Respiration of 2,4,6-trinitrotoluene by Pseudomonas sp. strain JLR11.

Under anoxic conditions Pseudomonas sp. strain JLR11 can use 2,4, 6-trinitrotoluene (TNT) as the sole N source, releasing nitrite from the aromatic ring and subsequently reducing it to ammonium and incorporating it into C skeletons. This study shows that TNT can also be used as a terminal electron acceptor in respiratory chains under anoxic conditions by Pseudomonas sp. strain JLR11. TNT-dependent proton translocation coupled to the reduction of TNT to aminonitrotoluenes has been observed in TNT-grown cells. This extrusion did not occur in nitrate-grown cells or in anaerobic TNT-grown cells treated with cyanide, a respiratory chain inhibitor. We have shown that in a membrane fraction prepared from Pseudomonas sp. strain JLR11 grown on TNT under anaerobic conditions, the synthesis of ATP was coupled to the oxidation of molecular hydrogen and to the reduction of TNT. This phosphorylation was uncoupled by gramicidin. Respiration by Pseudomonas sp. strain JLR11 is potentially useful for the biotreatment of TNT in polluted waters and soils, particularly in phytorhizoremediation, in which bacterial cells are transported to the deepest root zones, which are poor in oxygen.

Neurologic evaluation in thoracolumbar vertebral fractures.

A standardized and universal neurologic evaluation is of essential importance in defining the prognosis, treatment, and long-term results in patients with spinal cord injury. The American Spinal Injury Association was the first to publish a neurologic classification in 1992. This classification, which was subsequently submitted to numerous revisions, was also adopted by the International Medical Society of Paraplegia and by the national scientific associations of many other countries. In Italy, too, the ASIA/IMSOP form has been accepted as a standard for neurologic evaluation, but with the exception of specialized centers, it is still not well-known. The authors critically analyze the international terminology and its translation, they define the technical aspects of the neurologic examination in relation to priorities, and to the specific goals. Finally, they report several thoughts deriving from clinical practice in the Spinal Unit of Florence.

Constitutive choline transport in Pseudomonas aeruginosa.

Pseudomonas aeruginosa has a choline uptake system which is expressed in bacteria grown in the presence of succinate and ammonium chloride as the carbon and nitrogen source, respectively. This system obeys Michaelis-Menten kinetics with an apparent Km value of 53 microM; its activity is not inhibited by high osmolarities in the medium but is partially inhibited by choline metabolites such as betaine and dimethylglycine.

In vivo construction of a hybrid pathway for metabolism of 4-nitrotoluene in Pseudomonas fluorescens.

Pseudomonas fluorescens 410PR grows on 4-nitrobenzoate but does not metabolize 4-nitrotoluene. The TOL pWW0 delta pm plasmid converts 4-nitrotoluene into 4-nitrobenzoate through its upper pathway, but it does not metabolize 4-nitrobenzoate. P. fluorescens 410PR(pWW0 delta pm) transconjugants were isolated and found to be able to grow on 4-nitrotoluene. This phenotype was stable after growth for at least 300 generations without any selective pressure. P. fluorescens 410PR(pWW0 delta pm) converted 4-nitrotoluene into 4-nitrobenzoate via 4-nitrobenzylalcohol and 4-nitrobenzaldehyde. 4-Nitrobenzoate was metabolized via 4-hydroxylaminobenzoate and finally yielded NH4+ and 3,4-dihydroxybenzoate, which was mineralized.

[Myelomeningocele: surgical treatment and results]. / Mielomeningocele: tratamento cirúrgico e resultados.

The authors made a retrospective study of 72 patients with myelomeningocele treated at a pediatric hospital in Rio de Janeiro, Brazil. 65 (90.2%) lesions involved the inferior segments of the spine and 87.5% of the patients needed shunt procedures for hydrocephalus control. Shunt infections in spina bifida cystica patients were more frequent than in any other group of hydrocephalic patients treated at the institution. There was a great incidence of postoperative complications such as wound infections and skin necrosis. 11 (15.3%) patients developed signs and symptoms related to Chiari II malformation. The overall mortality was 8.3%, and in most of the cases, ascribed to the Chiari malformation.

[Ambulatory follow-up of patients with myelomeningocele in a pediatric hospital]. / Acompanhamento ambulatorial de pacientes com mielomeningocele em um hospital pediátrico.

Myelomeningocele is the most common congenital malformation of the nervous system and despite its complexity and involvement of multiple organs is compatible with long survival. The peculiar characteristics of this malformation expose myelomeningocele patients to acute and chronic care problems with effects in quality of survival. In order to evaluate the quality of the follow-up of spina bifida patients in a pediatric hospital, the authors examined 54 patients attending the neurosurgical outpatient unity of a pediatric hospital. The lack of a multidisciplinary spina bifida clinic in Rio de Janeiro forced the patients to pursuit for complimentary medical and paramedical care outside the hospital with significant effects in the quality of survival. In consequence, only 25% of the patients were able to walk and community ambulation was nearly absent. Only 66.6% had a regular rehabilitation program and nearly 50% had routine orthopedics consultations. Almost half of the patients had no urological referral at all and 75% were incontinent, with recurrent urinary infections ranging 72.2%. The rates of neurosurgical complications were similar to those observed in the literature. We concluded that the quality of survival of patients with neural tube defects is strongly influenced by the adverse socio-economical conditions and the lack of a spina bifida multidisciplinary clinic.

Sensitivity of the Ottawa rules.

STUDY OBJECTIVE: To validate criteria predicting ankle and mid-foot fractures with 100% sensitivity. DESIGN: Prospective validation study SETTING: A 929-bed community teaching hospital with an annual census of 76,488 ED visits. PARTICIPANTS: Convenience sample of patients older than 18 years with acute ankle or midfoot injury. INTERVENTIONS: Radiography was performed in each patient received after pertinent history and physical examination findings were recorded. RESULTS: Five hundred seventy radiographs were obtained in 484 patients. Four hundred twenty-one were of the ankle, and 149 were of the foot. There were 93 ankle fractures and 29 midfoot fractures, giving a fracture yield of 22.1% for ankle films and 19.5% for foot films. Decision rules had sensitivity of 94.6% and specificity of 15.5% for ankle fractures and sensitivity of 93.1% and specificity of 11.5% for midfoot fractures. Prospective criteria failed to predict fracture in five of the ankle group and two of the midfoot group. Physicians predicting fracture solely on the basis of clinical suspicion had a sensitivity of 69% in ankle injuries and 76% in midfoot injuries. CONCLUSION: We were unable to validate with 100% sensitivity the Ottawa rules predicting ankle and midfoot fractures. However, the Ottawa rules were more sensitive than clinical suspicion alone.

[Factors affecting recurrence after surgical treatment of Crohn disease]. / Fattori influenzanti la recidiva dopo trattamento chirurgico del morbo di Crohn.

Surgery does not cure Crohn's disease, but only its complications, as the recurrence rate that requires a new intervention is 6% per year. The resections performed by the surgeon should be as limited as possible, in order to avoid the consequent malabsorption. The identification of two forms of Crohn's disease, with different aggressiveness, has found that the stricturoplastic is an encouraging way of treatment for those forms with a prevalent stenotic component. A lot of studies have evaluated the relationships between recurrences and resections on margins microscopically free or affected by the disease. The aim of this study was a retrospective verification of the influence of any possible microscopical residue of the disease on the recurrence rate, evaluating whether the two different forms of aggressiveness of the disease (presence of stenosis or fistula) can influence the rate and precocity of the recurrence onset. In 37 patients operated for the first time of ileal or ileocolic resection, the overall recurrence rate was 18.9%; neither the presence of microscopically affected margins nor the presence of fistulas or stenosis has showed to have an influence on the onset of the recurrences. The only data that emerged is a greater precocity of the onset of recurrence in those patients whose disease was characterised by the presence of enteric fistulas. The forms in which fistulas and perforations were evident showed a recurrence rate not significantly higher than that of forms with stenosis only, but the period of time free from the disease was notably longer for the latter. In the end, patients in which typical granulomas were present showed a recurrence rate of just 9%, compared to 23% of patients in which granulomas were absent. MATERIALS AND METHODS. From 1980 through 1992, 61 patients affected by Crohn's disease were operated. There were 39 men and 22 women (mean age: 40.4 years). The mean length of the follow-up was 55.5 months. It was the first operation for 43 patients, while 9 had already undergone surgery in other hospitals; 9 patients showed anorectal complications. The operations performed on the patients for the first time have been ileal resection in the following localizations: duodenum-jejunum 4, jejunum and ileal 34, colic 5; the recurrences treated have been ileal-jejunum in 7 cases and colic in 2. In 2 cases of recurrence a stricturoplastic has been performed. RESULTS. The operative mortality was of 3 patients: 2 due to sepsis for anastomotic dehiscence and 1 to systemic mycosis. Four postoperative fistulas were observed. Recurrence of the disease occurred in 13 patients (26.5%), specifically in 21.4% of the patients operated for the first time and in 57.1% of those that were operated for recurrences. DISCUSSION AND CONCLUSIONS. In the treatment of Crohn's disease, it is important to identify any possible group with high risk of recurrence in order to undertake an appropriate medical prophylaxis. The results concerning the presence of microscopical disease on the resection margins are today still controversial. Some groups of authors prefer wide resection margins, some others are in favour of restricted resections. Our considerations let us assert that in those patients in which the resections have been performed on margins with microscopic presence of the disease, the interval before the recurrence occurs is not significantly shorter than that of patients with free margins. But the patients suffering from Crohn's disease with fistulae, probably need medical post-operative therapy to delay recurrences onset.

Carnitine resembles choline in the induction of cholinesterase, acid phosphatase, and phospholipase C and in its action as an osmoprotectant in Pseudomonas aeruginosa.

The present study demonstrates that under conditions of iso or hyperosmolarity, P. aeruginosa utilized carnitine as the carbon, nitrogen or carbon and nitrogen sources. As occurred in the case of choline, the bacteria synthesized cholinesterase (ChE), acid phosphatase (Ac.Pase) and phospholipase C (PLC) under any of these conditions and in the presence of high or low Pi concentrations. Carnitine acted as an osmoprotectant when the cells were grown in the presence of preferred carbon and nitrogen sources and high NaCl concentrations. Under these conditions the three enzyme activities were not produced. The osmotically stressed bacteria grown under any of the above conditions accumulated betaine. Its presence indicated that carnitine may be metabolized by P. aeruginosa to produce betaine which could account for the induction of the three enzyme activities or its action as an osmoprotectant. The phosphatidylcholine encountered in the host cell membranes allows the bacteria to obtain free choline by the coordinated action of PLC and Ac.Pase. Since the consequence of this action may be cell disruption, the increase of free carnitine in the natural environment of the bacteria is also possible. These two compounds, choline and carnitine, acting in conjunction or separately, may increase the production of PLC and Ac.Pase activities by P. aeruginosa and thus enhance the degradative effect upon the host cells.

A simple and reliable method for the purification of Pseudomonas aeruginosa phospholipase C produced in a high phosphate medium containing choline.

1. Pseudomonas aeruginosa phospholipase C from culture supernatants of bacteria grown in high-Pi basal salt medium with choline, as the sole carbon and nitrogen source, was purified by precipitation with 70% saturation ammonium sulfate in the presence of celite. 2. The PLC activity was eluted of this mixture by the use of a reverse gradient of 70-0% ammonium sulfate. 3. The peak containing the PLC activity revealed a single protein after SDS-PAGE. 4. The method could also be applied to purify PLC produced in a low-Pi complex medium. The resultant preparation was not homogeneous. 5. The molecular weight for both PLC preparations was about 70 kDa. 6. Both PLC used phosphatidylcholine and sphingomyelin as substrates, displayed hemolytic activity an exhibited an apparent KM of 25 mM for p-nitrophenylphosphorylcholine. 7. They were not inhibited by 1% sodium deoxycholate but were 30% inhibited by 1% Triton X-100. 8. 2% sodium dodecylsulfate and 1% tetradecyltrimethylammonium bromide inhibited the PLC from the HP1-BSM plus choline but not the enzyme from the LP1-CM.

[Effect of dobutamine on the transport and consumption of oxygen during general anesthesia with enflurane]. / Efecto de la dobutamina sobre el transporte y consumo de oxígeno durante la anestesia general con enflurano.

We undertook a comparative study on the effects of dobutamine and placebo on hemodynamic parameters, O2 transport (DO2), O2 consumption (VO2, and acid lactic production during peripheral arterial surgery under general anesthesia with enflurane at a constant inspiratory concentration (1.2%). This study involved 18 patients older than 65 years (9 patients allocated in the dobutamine group and 9 in the placebo group). The hemodynamic course was monitorized by means of noninvasive methods such as the aortic output measured by continuous transoesophageal echo-Doppler. After introduction of enflurane into the circuit aortic output decreased by 39% in dopamine group and 36% in placebo. Total systemic vascular resistances increased by 52% in dopamine and by 48% in placebo treatment. DO2 showed a decrease of 39% in dopamine and 38% in placebo group. There were no appreciable differences in VO2 among the two groups. Recovery of hemodynamic parameters and DO2 was only observed in the dopamine group when the drug was perfused at a rate of 4 +/- 1.2 micrograms/kg/min. Dobutamine induced a transient increase of VO2 up to 225% of the baseline value. During the postanesthetic period VO2 and blood acid lactic were significantly higher in the dopamine than in the placebo group (192%, p less than 0.01 and 33%, p less than 0.05, respectively). The course of hemodynamic parameters, DO2, VO2, and blood acid lactic of dobutamine group appear to demonstrate that dobutamine perfusion reverts myocardial depression and improves cellular perfusion during general anesthesia with enflurane.