Nematicidal activity of essential oils and volatiles derived from Portuguese aromatic flora against the pinewood nematode, Bursaphelenchus xylophilus.

Twenty seven essential oils, isolated from plants representing 11 families of Portuguese flora, were screened for their nematicidal activity against the pinewood nematode (PWN), Bursaphelenchus xylophilus. The essential oils were isolated by hydrodistillation and the volatiles by distillation-extraction, and both were analysed by GC and GC-MS. High nematicidal activity was achieved with essential oils from Chamaespartium tridentatum, Origanum vulgare, Satureja montana, Thymbra capitata, and Thymus caespititius. All of these essential oils had an estimated minimum inhibitory concentration ranging between 0.097 and 0.374 mg/ml and a lethal concentration necessary to kill 100% of the population (LC(100)) between 0.858 and 1.984 mg/ml. Good nematicidal activity was also obtained with the essential oil from Cymbopogon citratus. The dominant components of the effective oils were 1-octen-3-ol (9%), n-nonanal, and linalool (both 7%) in C. tridentatum, geranial (43%), neral (29%), and ß-myrcene (25%) in C. citratus, carvacrol (36% and 39%), γ-terpinene (24% and 40%), and p-cymene (14% and 7%) in O. vulgare and S. montana, respectively, and carvacrol (75% and 65%, respectively) in T. capitata and T. caespititius. The other essential oils obtained from Portuguese flora yielded weak or no activity. Five essential oils with nematicidal activity against PWN are reported for the first time.

San Diego pelvic osteotomy in patients with closed triradiate cartilage.

PURPOSE: A San Diego pelvic osteotomy is frequently used as part of the surgical treatment of neuromuscular patients with hip displacement. This short-term follow-up study assesses the effectiveness of this osteotomy in patients with closed triradiate cartilage compared with open triradiates. METHODS: Retrospective review of 43 patients (44 hips) with neuromuscular hip disease who underwent a San Diego pelvic osteotomy to correct hip displacement. In all 24 hips had open triradiate cartilage at the time of surgery and 20 hips were closed. Centre edge angle (CEA), acetabular angle (AA) and Reimer's index (RI) were recorded from preoperative, postoperative and the latest follow-up radiographs and statistical analysis was performed. RESULTS: CEA improved by 39° (7° to 69°) in the open group and 30° (9° to 80°) in the closed group from preoperative radiographs to postoperative, with no significant difference in improvement between groups (p = 0.084). There was no significant difference in AA between the open (improved by 11° (3° to 23°)) and closed (improved by 10° (4° to 21°)) groups (p = 0.65). RI improved from 61% to 11% in the open group and 51% to 12% in the closed group. There was no statistically significant difference between groups in RI at preoperative, immediate postoperative and final follow-up radiographs. At latest follow-up, CEA, AA and RI all remained relatively stable in both groups. CONCLUSIONS: San Diego pelvic osteotomy is equally effective in improving radiographic parameters in neuromuscular patients with both open and closed triradiate cartilage. This study challenges the notion that closed triradiate cartilage is a contraindication to a San Diego pelvic osteotomy. LEVEL OF EVIDENCE: III.

Unsteady walking as a symptom in type 2 diabetes mellitus: independent association with depression and sedentary lifestyle and no association with diabetic neuropathy.

The purpose of this study was to look at the determinants of the unsteady walking (UW) symptom in patients with type 2 diabetes mellitus (T2DM) by defining if UW and/or the Diabetic Neuropathy Symptoms Score (DNSS) are associated with positive scores in Beck's Depression Inventory (BDI) and with a positive Michigan Neuropathy Screening Instrument score (MNSI). We evaluated 203 T2DM patients without visible gait disturbances. They were divided into UW (+) and UW (-) or DNSS (+) and DNSS (-) according to symptoms. We found a prevalence of 48.3% for UW (+) and of 63% for DNSS (+) in our sample. In univariate analysis, the presence of UW was significantly associated with waist circumference (P=0.024), number of comorbidities (P=0.012), not practicing physical exercise (P=0.011), positive BDI score (P=0.003), presence of neuropathic symptoms by the MNSI questionnaire (P<0.001), and positive diabetic neuropathy screening by MNSI (P=0.021). In multivariate analysis, UW (used as a dependent variable) was independently associated with a positive BDI score (P<0.001; 95%CI=1.01-1.03), T2DM duration (P=0.023; 95%CI=1.00-1.03), number of co-morbidities (P=0.032; 95%CI=1.01-1.37), and a sedentary lifestyle (P=0.025; 95%CI=1.06-2.5). The UW symptom and a positive DNSS are more closely related to a positive score for depression than to presence of neuropathy in T2DM.

Quantitative autoradiography of Na+-dependent [3H]L-aspartate binding to L-glutamate transporters in rat brain: structure-activity studies using L-trans-pyrrolidine-2,4-dicarboxylate (L-t-PDC) and 2-(carboxycyclopropyl)-glycine (CCG).

Sodium-dependent binding of [3H]L-aspartate was studied in thaw-mounted horizontal sections of fresh-frozen (i.e. not fixed) rat brain. After the incubation with [3H]L-aspartate, the sections were exposed against a 3H-sensitive film and the resulting autoradiograms were evaluated by quantitative densitometry. Effects of several inhibitors were examined and their potency expressed as IC50 and nH. Together with previously published data, the present study supports the view that [3H]L-aspartate binding to fresh-frozen sections of rat brain represents interaction of the radioligand with the substrate-binding sites on glutamate transporters. The most potent inhibitors were (2S,3S,4R)-2-(carboxycyclopropyl)-glycine (L-CCG III) and (2S,4R)-4-methylglutamate. In contrast, L-anti,endo-3,4-methanopyrrolidine dicarboxylate (L-a,e-MPDC) was about an order of magnitude less potent. Only subtle regional variations in the characteristics of inhibitors of [3H]L-aspartate binding were detected. It is not certain whether these differences reflect regional variations in the distribution of individual glutamate transporters or regional peculiarities in their pharmacological characteristics. In particular, (2S,4R)-4-methylglutamate, shown previously to differentiate between GLT-1 (principal glutamate transporter in the forebrain) and GLAST (expressed mainly in the cerebellum), did not strongly differentiate between the binding of [3H]L-aspartate in forebrain and cerebellum. Computer-assisted molecular modelling using selected glutamate analogues with restricted conformation (L-trans-pyrrolidine-2,4-dicarboxylate and four isomers of 2-(carboxycyclopropyl)-glycine: L- and D-CCG I, L-CCG III and L-CCG IV) identified at least one area of unfavourable steric interaction. We conclude that the quantitative autoradiographic studies using [3H]L-aspartate or other transporter-specific ligands, will be a useful tool to study the pharmacology of substrate binding sites on glutamate transporters in the mammalian brain in situ.

Inhibition of [3H]CGP 39653 binding to NMDA receptors by a P2 antagonist, suramin.

Suramin, which has been used for more than 50 years as an anthelmintic in humans, has recently been shown to inhibit P2 purinoceptors in the CNS and to block glutamate-evoked excitatory potentials in hippocampus. We now report that suramin inhibits (IC50 approximately 100 microM) the binding of [3H]CGP 39653, a ligand specific for NMDA-type glutamate receptors in brain. Consequently, suramin at concentrations used sometimes in physiological experiments may act as both P2x and NMDA antagonist. Since both NMDA receptors and P2x-subtype of purinoceptors form Ca(2+)-selective channels, suramin may block Ca2+ entry into neurones depolarized by either ATP, L-glutamate or any other neurotransmitter or drug acting via P2x or NMDA receptors.

Effect of post-mortem delay on density of tachykinin receptors in rat peripheral tissues.

The effect of post-mortem delay on the affinity and density of tachykinin NK1 and NK2 receptors was examined in the rat submandibular gland and gastric fundus, respectively, using saturation binding studies with the radioligands [125I]Bolton-Hunter [Sar9, Met(O2)11]SP and [125I][Lys5, Tyr(I2)7, MeLeu9, Nle10]NKA(4-10). For NK1 receptors, no significant changes were seen in either Kd (control 375 +/- 35 pM, n = 5; 32 h post-mortem 390 +/- 59 pM, n = 5) or Bmax (control 96 +/- 16 fmol/mg protein, n = 5; 32 h post-mortem 62 +/- 10 fmol/mg protein, n = 5). For NK2 receptors, no alterations were seen up to 16 h post-mortem. However, significant (p < 0.001) changes were seen at 32 h post-mortem (n = 4), where values for Kd were increased (3.0 +/- 0.2 nM) and those for Bmax were reduced (42 +/- 5.9 fmol/mg protein), relative to control (Kd = 1.3 +/- 0.2 nM; Bmax = 208 +/- 30 fmol/mg protein, n = 5). These changes are probably related to observed histological deterioration. This study demonstrates the stability of tachykinin receptors in these peripheral tissues and indicates the suitability of post-mortem tissue as a valid control in future tachykinin receptor studies.

Binding characteristics of endothelin ET(A) receptors in normal and post-mortem rat lung.

In control lung homogenates, optimal specific binding of [(125)I]endothelin-1 and minimal filter binding was achieved using 50 microg/ml bacitracin, 30 microM phenylmethylsulphonyl fluoride (PMSF) and 10 mM EDTA. In post-mortem tissue (8, 16, and 32 h), no significant changes were seen in ET(A) receptor affinity (K(d)) or number (B(max)): control and 32 h K(d) = 309 +/- 75, 225 +/- 32 pM and B(max) = 173 +/- 42, 185 +/- 17 fmol/mg protein, respectively. Autoradiographic binding sites for [(125)I]endothelin-1 were densely expressed on bronchiolar smooth muscle and parenchyma with moderate binding on epithelium and blood vessels. Histologic sections of post-mortem lung showed minimal deterioration of structures expressing ET(A) binding sites. Hence the ET(A) receptor is stable in the rat lung for up to 32 h post-mortem.

Autoradiographic studies indicate regional variations in the characteristics of L-glutamate transporters in the rat brain.

Quantitative autoradiography of [3H]L-aspartate binding in thaw-mounted sections of rat brain has shown that L-trans-pyrrolidine-2,4-dicarboxylate and D-threo-3-hydroxyaspartate but not DL-2 aminoadipate strongly interacted with the binding sites while dihydrokainate, kainate and beta-aminoadipate produced only weak effects. The potency of inhibitors did not vary from one region to another in the telencephalon (neocortex, hippocampus and neostriatum) but, D-threo-3-hydroxyaspartate, L-trans-pyrrolidine-2,4-dicarboxylate, kainate and dihydrokainate inhibited [3H]L-aspartate binding in the cerebellar cortex less potently than that in the forebrain. Characteristics of the known excitatory amino acid transporters can, in part, explain the present results but contributions from additional transporter molecules to the heterogeneity of [3H]L-aspartate binding sites cannot be ruled out.

Strategies for studies of neurotoxic mechanisms involving deficient transport of L-glutamate: antisense knockout in rat brain in vivo and changes in the neurotransmitter metabolism following inhibition of glutamate transport in guinea pig brain slices.

This communication briefly reviews characteristics of glutamate transport in the central nervous system and is involved in the aetiology of slow neurodegenerative diseases. Data in the literature suggest that antisense oligonucleotides targeted against glutamate transporters and administered in vivo over a period of days could be used to test the hypothesis. Data from our laboratory have indicated that single intraventricular doses of antisense oligonucleotides can also results in significant reductions in the numbers of substrate binding sites associated with glutamate transporters and may even cause subtle changes in their characteristics. In order to study metabolism in brain tissue, we have used 13C-nuclear magnetic resonance spectroscopy to analyse extracts of slices of guinea pig cerebral cortex exposed to glutamate transport inhibitor L-anti,endo-methanopyrrolidine dicarboxylate (L-a,e-MPDC). The results have shown-for the first time in an experimental model that preserves the relationship between glia and neurones within the context of brain tissue-that inhibition of L-glutamate transport can exert a significant influence on neurotransmitter-related metabolism. These findings suggest that metabolic disturbances caused by deficient glutamate transport could play a significant role in the death of neurones under pathological conditions in vivo.

Fractures of the distal tibial epiphysis in adolescence.

We studied the cases of nine adolescent patients with a Salter-Harris Type-III fracture of the lateral portion of the distal tibial epiphysis (sometimes called the juvenile Tillaux fracture) and eight patients with a triplane fracture of the distal tibial epiphysis. The mechanism of both injuries was postulated to be external rotation of the foot on the leg. There were both two-fragment and three-fragment triplane fractures in the series. The three-fragment triplane fractures occurred in younger patients, and surgical treatment was more frequently administered. The pattern of closure of the distal tibial growth plate in the adolescent strongly influenced the type of fracture.

Rotational deformities of the lower limb in myelomeningocele. Evaluation and treatment.

We studied the results of surgical treatment for rotational deformities of the lower extremity in fifty children. The preoperative rotational deformities were subdivided as follows: (1) external rotation deformity of the hip (nine hips), (2) external tibial torsion (twenty-nine limbs), and (3) toeing-in gait (thirty-one limbs). These were further analyzed as to imbalance between the forces of the medial and lateral hamstrings, that imbalance associated with fixed internal tibial torsion, and isolated internal tibial torsion. The average length of follow-up was four years and six months. For the external rotation deformity of the hip we performed a derotation osteotomy with internal fixation at the subtrochanteric level. That procedure led to eight good results and one fair result. The external tibial torsion was treated by a derotation osteotomy of the tibia and fibula distally, with correction of the valgus deformity of the ankle if that was more than 10 degrees. In twenty patients a good result was obtained. For the toeing-in gait associated with a medial-lateral hamstring imbalance, the semitendinosus was transferred to the biceps and the head of the fibula. Good results were seen in ten of the fourteen patients. In twelve limbs a derotation osteotomy of the distal ends of the tibia and fibula was done alone, with eight good and fair results. In five patients a simultaneous semitendinosus transfer and derotation osteotomy was used. All had a good result. The over-all rate of good and fair results in the entire series was 79.6 per cent.

Selective posterior rhizotomy and soft-tissue procedures for the treatment of cerebral diplegia.

The results of selective posterior rhizotomy in fifty patients (group I) and of soft-tissue procedures in fifty patients (group II), all 100 of whom had cerebral diplegia and were seen in a private office, were reviewed retrospectively. No effort was made to randomize the treatment, as the selection criteria for the two procedures are different. We evaluated the range of motion and the ability and quality of walking preoperatively and postoperatively as well as the need for additional operative intervention in the two groups. The average age of the patients in both groups was five years (range, three to twelve years in group I and one to thirteen years in group II). The average duration of follow-up in both groups was four years (range, one to six years in group I and one to seven years in group II). Thirty-two patients (64 percent) in group I and forty-one patients (82 percent) in group II were able to walk independently at the latest follow-up examination. Both groups had an over-all improvement in the ranges of abduction of the hips and dorsiflexion of the ankles, a decrease in the flexion contractures of the hips, and more normal popliteal angles; however, with the numbers available, there were no significant differences in these measurements between the two groups at the 0.05 percent confidence level. Despite the overall improvement in range of motion, thirty-one patients in the rhizotomy group subsequently had soft-tissue releases, and twenty-two patients in the soft-tissue-release group had additional operative intervention.

Musculoskeletal problems in association with cloacal exstrophy.

The records of all thirteen patients for whom a diagnosis of cloacal exstrophy had been recorded in our hospitals were analyzed for evidence of musculoskeletal problems. All thirteen patients had spina bifida, four had congenital scoliosis, two had congenital kyphosis, and three had non-congenital scoliosis. All had a lipomeningocele, and eleven had paralysis of the lower extremities. Hydrocephalus and its associated problems were not found, but tethered-cord syndrome was diagnosed in eleven patients. Persistent diastasis of the symphysis pubis was found in all patients. Abduction and external rotation of the hips were more than normal. Mild dysplasia was seen in six of the twenty-six hips. Deformities of the foot were common, and twelve feet had been operated on for correction. Recurrent equinovarus deformity of the foot was associated with tethered-cord syndrome in two patients.

The natural history of congenital kyphosis in myelomeningocele. A review of 51 children.

The progression of congenital lumbar kyphosis in myelomeningocele is a well-known problem, but rates of progression are not well documented in the literature. Fifty-one children with congenital kyphosis and myelomeningocele were followed for an average of 4.8 years. Minimum follow-up was 1 year. Group I (35 patients) had initial radiographs at 1 year of age or less. Group II (16 patients) had radiographs taken after the age of 1 year. Curves less than or equal to 90 degrees in Group I progressed 7.7 degrees/yr; those greater than 90 degrees progressed 12.1 degrees/yr. Curves less than or equal to 90 degrees and greater than 90 degrees progressed at similar rates, regardless of initial curve magnitude: 6.4 degrees/yr and 6.7 degrees/yr, respectively. No correlation existed among the rate of curve progression, the frequency of shunt revisions, or the presence of vertebral anomalies, aside from the dysraphism.

Evaluation of treatment of hip subluxation in myelomeningocele by intertrochanteric varus derotation femoral osteotomy.

A retrospective study of 27 subluxed hips was carried out in 18 patients with myelomeningocele treated by varus derotation intertrochanteric osteotomy. Twenty-three of the 27 hips were stable at follow-up review. The causes of failure were related to the presence of pelvic obliquity secondary to scoliosis or to a dysplastic acetabulum (acetabular index above 25 to 30 degrees). Transfer of the iliopsoas tendon was not found to be necessary in order to achieve stable reduction.

Strategies of hip management in myelomeningocele: to do or not to do.

Paralytic hip dislocation in myelomeningocele is common and can be a complicated problem. This review summarizes results of surgical techniques employed in this patient population in order to achieve and maintain reduction of paralytic hip dislocations. This review also examines the controversial question of whether or not hip surgery in patients with myelomeningocele provides improved functional results. Finally this paper suggest appropriate goals and recommendations for treatment of the paralytic hip dislocation in myelomeningocele.

Describing phytotoxic effects on cumulative germination.

Phytotoxic studies strongly depend on evaluation of germination responses, which implies the need for adequate procedures to account for distinct aspects of the germinative process. For this, indices, comparisons among treatments at various times, and model fitting have been proposed. The objective of this work is to compare the three approaches and select the one providing the greatest insight and precision. Speed of germination, speed of accumulated germination, the coefficient of the rate of germination, comparisons at each determination time, including final germination, and the parameters of the Weibull function were examined. The Weibull function proved the best approach to describe the germination process, providing not only the same type of information about the speed of germination, with greater precision, but also additional information about the initiation and shape of the germination response curve.

Valgus deformity of the ankle joint: pathogenesis of fibular shortening.

An abnormal shortening of the fibula, leading to a valgus deformity of the ankle joint, is a common finding in the paralytic ankle. This article analyzes 173 ankles in which shortened fibulae were present. Analysis of several factors that interfere with fibular growth showed that soleus strength and anatomical continuity of the fibula are important factors in the normal fibular growth. Any factor that interferes with the normal balance of forces at the distal fibula physis can cause an abnormal shortening of the fibula and lead to valgus deformity of the ankle.