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ims/hypothesis: Diabetes is considered the leading cause of neuropathies in developed countries. Dysfunction of nerve growth factor (NGF) production and/or utilisation may lead to the establishment of diabetic neuropathies. Electroacupuncture has been proved effective in the treatment of human neuropathic pain as well as in modulating NGF production/activity. We aimed at using electroacupuncture to correct the development of thermal hyperalgesia and the tissue alteration of NGF and sensory neuromodulators in a rat model of type 1 diabetes. Methods: Adult rats were injected with streptozotocin to induce diabetes and subsequently treated with low-frequency electroacupuncture for 3 weeks. Variation in thermal sensitivity was studied during the experimental course. Hindpaw skin and spinal cord protein content of NGF, NGF receptor tyrosine kinase A (TrkA), substance P (SP), transient receptor potential vanilloid 1 (TRPV1) receptor and glutamic acid decarboxylase-67 (GAD-67) were measured after electroacupuncture treatments. The skin and spinal cord cellular distribution of TrkA was analysed to explore NGF signalling. Results: Early after streptozotocin treatment, thermal hyperalgesia developed that was corrected by electroacupuncture. The parallel increases in NGF and TrkA in the spinal cord were counteracted by electroacupuncture. Streptozotocin also induced variation in skin/spinal TrkA phosphorylation, increases in skin SP and spinal TRPV1 and a decrease in spinal GAD-67. These changes were counteracted by electroacupuncture. Conclusions/interpretation: Our results point to the potential of electroacupuncture as a supportive therapy for the treatment of diabetic neuropathies. The efficacy of electroacupuncture might depend on its actions on spinal/peripheral NGF synthesis/utilisation and normalisation of the levels of several sensory neuromodulators. [ABSTRACT FROM AUTHOR]

Summary: An increasing number of researchers of the metabolic syndrome assume that many mechanisms are involved in its complex pathophysiology such as an increased sympathetic activity, disorders of the hypothalamo–pituitary–adrenal axis, the action of chronic subclinical infections, proinflammatory cytokines, and the effect of adipocytokines or psychoemotional stress. An increasing body of scientific research in this field confirms the role of the neurotrophins and mastocytes in the pathogenesis of inflammatory and immune diseases. Recently it has been proved that neurotrophins and mastocytes have metabotrophic effects and take part in the carbohydrate and lipid metabolism. In the early stage of the metabolic syndrome we established a statistically significant increase in the plasma levels of the nerve growth factor. In the generalized stage the plasma levels of the neutrophines were statistically decreased in comparison to those in the healthy controls. We consider that the neurotrophin deficit is likely to play a significant pathogenic role in the development of the metabolic anthropometric and vascular manifestations of the generalized stage of MetSyn. We suggest a hypothesis for the etiopathogenesis of the metabolic syndrome based on the neuro-immuno-endocrine interactions. The specific pathogenic pathways of MetSyn development include: (1) increased tissue and plasma levels of proinflammatory cytokines Interleukin-1(IL-1), Interleukin-6 (IL-6 ) and tumor necrosis factor – α (TNF-α) caused by inflammatory and/or emotional distress; (2) increased plasma levels of neurotrophin – nerve growth factor (NGF) caused by the high IL-1, IL-6 and TNFα levels; (3) high plasma levels of NGF which enhance activation of: the autonomous nerve system – vegetodystonia (disbalance of neurotransmitters); Neuropeptide Y (NPY) – enhanced feeding, obesity and increased leptin plasma levels; hypothalamo–pituitary–adrenal axis – increased corticotropin-releasing hormone (CRH) and cortisol (hormonal disbalance); immune cells – increased number and degranulation of mastocytes (MC) – immunological disbalance; (4) as a result of 1–3 insulin resistance is exhibited leading to diabetes mellitus. The hypothesis is confirmed by results obtained after 6-month nonsteroid anti-inflammatory treatment of patients with MetSyn. These results are reported in a separate publication. [Copyright &y& Elsevier]

The aim of the present work was to relate age-related individual differences in cognitive function with behavioural strategies employed in social and non-social challenges. To this purpose, the behaviour of adult (5-month-old) and middle-aged (13-month-old) CD-1 mice was scored in the social interaction, plus-maze, Morris water maze (MWM) and open-field tests. In addition, brain levels of nerve growth factor and brain-derived neurotrophic factor (BDNF) were analysed and correlated with the behaviours scored. Compared to adults, middle-aged mice showed greater anxiety in both non-social and social situations, spending less time in the open arms of the plus-maze and performing more freezing behaviour in response to aggression. Based upon their behaviour in the social interaction test, adult and middle-aged subjects were classified as dominant or subordinate and their behaviour in the open field, plus-maze and MWM tests subjected to factor analysis, taking into account age and social status. Results highlighted meaningful differences in exploratory strategies as a function of social status only in middle-aged subjects. In particular, middle-aged dominants were, overall, more explorative than same-aged subordinates, spending less time in peripheral areas and approaching more readily a novel object. Interestingly, in middle-aged mice, superior performance in the MWM task was associated with exploratory strategies exploited by dominants. At adulthood, BDNF hippocampal levels, but not specific behaviours, were positively correlated with the ability to learn a spatial task. Overall, data indicate that, in middle-aged subjects individual differences in exploratory strategies, rather than neurotrophin levels, are able to predict the degree of impairment in a spatial learning task. [ABSTRACT FROM AUTHOR]

Oestadiol valerate (EV)-induced polycystic ovaries (PCO) in rats cause anovulation and cystic ovarian morphology. Denervation of ovarian sympathetic nerves restores ovulatory disruption. In the present study, we determined whether 5 weeks of voluntary exercise influence ovarian morphology and the expression of sympathetic markers in the EV-induced PCO rat model. The effect of exercise on (i) ovarian morphology; (ii) mRNA and protein expression of nerve growth factor (NGF); and (iii) mRNA and number of ovarian-expressing cells for the NGF receptor (p75 neurotrophin receptor) and the α1a-, α1b-, α1d- and β2-adrenergic receptors (ARs) in rats with EV-induced PCO was evaluated. PCO was induced by a single i.m. injection of EV, and controls were injected with oil alone in adult cycling rats. The rats were divided into four groups: (i) control (oil); (ii) exercise group (oil + exercise); (iii) a PCO group (EV); and (iv) a PCO exercise group (EV + exercise). The exercise and PCO exercise groups ran voluntarily for 5 weeks in computer-monitored wheels placed in the cages where they were housed. The results obtained indicated that ovarian morphology was almost normalised in the PCO exercise group; NGF mRNA and protein concentrations were normalised in the PCO exercise group; high numbers of NGF receptor expressing cells in PCO ovaries were lowered by exercise; and the number of immunopositive cells of the different AR subtypes were all reduced after exercise in the PCO group, except for the α1b- and β2-AR whereas the mRNA levels were unaffected, indicating transcriptional regulation. In conclusion, our data indicate a beneficial effect of regular exercise, as a modulator of ovarian sympathetic innervation, in the prevention and treatment of human PCOS. [ABSTRACT FROM AUTHOR]

It has been shown that lung mast cells, eosinophils, and fibroblasts are receptive to the action of nerve growth factor (NGF) and that NGF is released in to the bloodstream of subjects affected by allergic inflammatory response. The role of NGF in lung inflammatory disorders is unclear because there is evidence suggesting that NGF can be involved in both proinflammatory and anti-inflammatory responses. Lung fibroblasts play a marked role in inflammation. In this study we investigated the effect of NGF, interleukin 1β (II-1β), and dexamethasone (DEX) on human lung fibroblasts in vitro. We found that II-1β, but not NGF. promotes fibroblasts' survival and that NGF stimulates trkA receptor expression, down regulates TFG-α, and has no effect on TNF-β immunoreactivity. Moreover. DEX exerts different effects on NGF release by fibroblasts pre-exposed to II-1β. Our findings suggest that the NGF released by lung flbroblast during inflammation is not associated with the increase of proinflammatory factors such as TNF-α and II-1β. [ABSTRACT FROM AUTHOR]

Nerve growth factor (NGF) and nerve growth factor receptor (NGFR) expressions have been found to be increased in sub-conjunctival scarring.The aim of this study was to investigate thein vitroeffects of NGF on some pro-fibrogenic properties of human conjunctival fibroblasts.Expression of NGF, trkANGFR and p75NTR on human fibroblasts grown from conjunctival biopsies and incubated for 2 or 6 days with NGF were evaluated by immunofluorescence, RT-PCR, flow cytometry and ELISA. The fibrogenic effect of NGF on conjunctival fibroblasts was investigated by evaluating their migration (wound model), proliferation ([3H]-thymidine incorporation), collagen production (3H]-proline incorporation), expression of alpha-smooth muscle actin (α-SMA) (cell surface ELISA) and contraction of 3D collagen gels.NGF induced the expression of p75NTR in the fibroblasts that constitutively expressed only trkANGF and increased the migration of wounded fibroblasts, but not their proliferation and collagen production. NGF induced the conversion of fibroblasts into myofibroblasts expressingα-SMA, and enhanced their contraction of a collagen matrix. Interestingly, chronic NGF treatment induced transforming growth factor-β1 (TGF-β1) production by fibroblasts, and following specific TGF-β neutralization, all the NGF-induced effects were completely abrogated.Our findings indicate that NGF, via TGF-β induction, is likely to be involved in the healing or fibrotic processes occurring in conjunctiva during some pathological conditions. [ABSTRACT FROM AUTHOR]

Perinatal asphyxia is a concern for public health and may promote subtle neuropsychiatric disorders. Anoxic insults to neonatal rats cause long-lasting neurobehavioral deficits. In the present study, we focussed on changes in emotional behaviors as a consequence of neonatal asphyxia in Wistar rats. Newborn pups (24 h after birth) underwent a single 30-min exposure to a 100% N2 atmosphere (or air). The offspring was tested for a) locomotor and exploratory activity with or without ad-amphetamine challenge (0, 1, or 2 mg/kg) on postnatal day (pnd) 15; b) social interactions and novelty seeking during adolescence; c) levels of the brain-derived neurotrophic factor (BDNF). In the open-field test (pnd 15), N2-exposed pups injected with the high (2 mg/kg) amphetamine dose exhibited reduced levels of locomotor hyperactivity, and a more marked involvement in stereotyped behaviors. Individual differences emerged in the locomotor response to the novelty-seeking test: two subgroups of rats (separated on the basis of the median value) showed either arousal/attraction or avoidance/inhibition in response to free-choice novelty. The N2-exposed group showed a more marked novelty-induced avoidance and inhibition. Time devoted to allogrooming and play-soliciting behaviors was reduced, whereas object exploration was increased. Levels of BDNF were reduced in the striatum of N2-exposed rats, suggesting poorer synaptic performance of dopamine pathways. In conclusion, these findings suggest an increased risk of developing social withdrawal, neophobia and behavioral stereotypies (common symptoms found in schizophrenia and autism) as a consequence of neonatal asphyxia in preterm humans. [ABSTRACT FROM AUTHOR]

Aim: To evaluate the expression of neurotrophic factors (nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF)) and their association with the clinical-radiological characteristics and outcome of children with viral and bacterial meningoencephalitis (ME).Methods: Prospective observational clinical study performed on 13 children with ME and 12 controls with non-inflammatory obstructive hydrocephalus. Neurotrophic factor levels in the cerebro-spinal fluid (CSF) and plasma were measured using an immunoenzymatic assay.Results: High levels of NGF and BDNF were demonstrated in all patients, while GDNF levels did not undergo significant variations. NGF expression in the CSF was higher in viral ME than in bacterial ME and was correlated with CSF cellularity (particularly mononuclear cells). BDNF expression in the CSF was higher in bacterial ME than in viral ME and was correlated with CSF cellularity and blood platelet count. No relationships were noted between CSF protein or serum C-reactive protein levels and the expression of neurotrophic factors. Regarding clinical and radiological features, elevated NGF/BDNF levels in the CSF correlated with higher incidence of seizures and prolonged comatose state and with specific radiological lesions. No correlation was found between NGF/BDNF levels and final outcome.Conclusions: The variations in neurotrophic factor levels may reflect an endogenous attempt at neuroprotection against biochemical and molecular changes during both viral and bacterial ME. The expression of these factors is likely to play a neuro-immunomodulatory or neurosurvival role in ME infections. [ABSTRACT FROM AUTHOR]

Background: The prevalence of pressure ulcers of the foot is a major health care problem in frail elderly patients. A pressure sore dramatically increases the cost of medical and nursing care, and effective treatment has always been an essential nursing concern. Management options for pressure ulcers include local wound care; surgical repair; and, more recently, topical application of growth factors.Objective: To examine the effects of topical treatment with nerve growth factor in patients with severe, noninfected pressure ulcers of the foot.Design: Randomized, double-blind, placebo-controlled trial.Setting: Teaching nursing home of Catholic University of the Sacred Heart, Italy.Patients: 36 persons with pressure ulcers of the foot.Intervention: 18 patients received nerve growth factor treatment, and 18 patients received only conventional topical treatment.Measurements: The course of the ulcers during follow-up was evaluated by tracing the perimeter of the wound onto sterile, transparent block paper and determining the stage.Results: At baseline, the treatment and control groups did not differ across demographic variables, clinical characteristics, and functional measures. The mean area (+/-SD) of the ulcers was 1012 +/- 633 mm2 in the treatment group and 1012 +/- 655 mm2 in the control group (P > 0.2). The average reduction in pressure ulcer area at 6 weeks was statistically significantly greater in the treatment group than in the control group (738 +/- 393 mm2 vs. 485 +/- 384 mm2; P = 0.034).Conclusion: Topical application of nerve growth factor may be an effective therapy for patients with severe pressure ulcers. [ABSTRACT FROM AUTHOR]

Stress is elicited by environmental, social or pathological conditions occurring during the life of animals and humans that determine changes in the nervous, endocrine and immune systems. In the present review, we present data supporting the hypothesis that stress-related events both in animal models and humans are characterized by modifications of endogenous nerve growth factor (NGF) synthesis and/or utilization. Stress inducing alteration in NGF synthesis and/or utilization appears to be more severe during neurogenesis and in early postnatal life. However, NGF endogenously released during stress may promote remodeling of damaged tissues following acute and/or chronic stressful events. [Copyright &y& Elsevier]

Schistosomiasis is a parasitic disease due to Schistosoma mansoni .Schistosome infection is known to induce granulomas not only in the spleen,bladder, liver and intestine but also in the brain and spinal cord resultingin severe neuropathological and psychiatric disorders though the interactionbetween Schistosoma mansoni infection and the nervous system hasreceived on the whole little attention. In the present review it has beendiscussed recent findings from experimental Schistosoma mansoni infectionin mouse nervous system. We show that brain granulomas are associated witha significant alteration in the constitutive levels of nerve growth factor(NGF), a trophic factor playing an essential role in nerve growth and differentiationand in preventing neuronal damages. Animals infected with schistosomes sufferedalso of increased pain sensitivity which was inhibited by TNF-α antibodyinjections and not by anti-NGF. These findings suggest that the neuropathologicaldysfunctions in neuroschistosomiasis may be linked to changes in the basallevels and/or activity of neurotrophic factors caused by local formation ofgranulomas. [ABSTRACT FROM AUTHOR]

While multiple growth factor, cytokines, and immune cells are identifiedin atherosclerotic lesions, as well as an essential nonneuronal function ofneurotrophins implicated in cardiovascular tissue development and in lipidand glucose metabolism, the role of the neurotrophins NGF and BDNF and alsothe adipokine leptin in human coronary atherosclerosis and related disorders,such as metabolic syndrome, remains unclear. Here we report that (i) boththe amount and the immunoreactivity of NGF was reduced and the expressionof p75NGF receptor and the number of mast cell increased in human atheroscleroticcoronary arteries (n = 12) compared with control specimens (n = 9) obtainedfrom autopsy cases, and (ii) NGF and BDNF plasma levels were reduced in patientswith metabolic syndrome (n = 23) compared with control subjects (n = 10).Also, in metabolic syndrome patients, a positive correlation between the plasmaleptin levels and the number of adipose tissue mast cells was found, suggestingthat leptin may be a novel adipoimmune mediator. Altogether, the results providethe first correlative evidence for the potential involvement of NGF, BDNF,leptin, and mast cells in human coronary atherosclerosis and metabolic syndrome,implying neuroimmune and adipoimmune pathways in the pathobology of thesecardiovascular disorders. [ABSTRACT FROM AUTHOR]

Objective: To determine the circulating levels of nerve growth factor (NGF), neuropeptide Y (NPY), and vasoactive intestinal peptide (VIP) in systemic sclerosis (SSc), and to correlate these levels with clinical and laboratory features.Methods: Forty four patients with SSc were evaluated for circulating NGF (immunoenzymatic assay), NPY and VIP (radioimmunoassay), anticentromere and antitopoisomerase I autoantibodies, lung disease (pulmonary function tests with carbon monoxide transfer factor (TLCO), ventilation scintiscan with 99mTc DTPA radioaerosol, high resolution computed tomography (HRCT), pulmonary pressure (echo colour Doppler)), heart disease (standard and 24 ECG, echocardiography), cutaneous involvement (skin score), joint involvement (evidence of tender or swollen joints, or both), peripheral nervous system (PNS) involvement (electromyography), rheumatoid factor, angiotensin converting enzyme (fluorimetric method), von Willebrand factor (ELISA), and erythrocyte sedimentation rate (ESR) (Westergren).Results: Circulating NGF levels in SSc were significantly increased compared with controls (p<0.00001) and significantly higher in the diffuse than in the limited subset of patients (p<0.01). Patients with articular disease had significantly higher levels of NGF. A significant indirect correlation between NGF levels and TLCO was detected (p<0.01), but no correlation was found between NGF and HRCT, DTPA, skin score, PNS involvement and angiotensin converting enzyme and von Willebrand factor levels, antitopoisomerase or anticentromere antibodies, and ESR. NGF levels increased progressively as the disease worsened. Similarly, VIP circulating levels were significantly increased in patients with SSc (p<0.001), whereas the increase of NPY levels did not reach statistical significance. However, both neuropeptides, following the same trend as NGF, increased as the disease worsened (skin score and lung disease).Conclusions: The increase of NGF and VIP in patients with SSc, the former in the diffuse subset of the disease, and in patients with prominent articular disease, may suggest a link between neurotransmitters and the disease pathogenesis. Neuropeptide circulating levels seem to increase only in patients with the most severe disease. [ABSTRACT FROM AUTHOR]

In this study we measured with a highly sensitive two-site immunoenzymatic assay the levels of nerve growth factor (NGF) in human optic nerve affected by multiple sclerosis (MS). The result of this study showed an elevated increase of NGF in the MS optic nerves, as compared to the level of NGF found in the optic nerves non-affected by this demyelinating disorder. Moreover, the results showed that the optic nerves also expressed trkA NGF receptor and NGFmRNA, most likely by oligodendrocytes, implying that NGF is locally produced and suggesting that the presence of NGF might be regulated by an autocrine mechanism. These and other ongoing studies on animal models indicate that altered NGF levels are among one of the early symptoms of these demyelinating diseases. The physiopathological role of NGF in the optic nerve during demyelinating disorders remains however to be defined. [ABSTRACT FROM AUTHOR]

Abstract Using adult mice infected with the trematode Schistosoma mansoni, we observed that this infection induces both thermal hyperalgesia and an increase in the levels of nerve growth factor in the paws. To explore the mechanism involved in peripheral hypersensitivity during chronic infection, mice were infected with 60 cercariae of S. mansoni and injected 17 weeks later with nerve growth factor, anti-nerve growth factor or with other molecules known to be associated with hyperalgesic processes. The results of these studies showed that antibodies against tumor necrosis factor-alpha, but not against nerve growth factor, reduce thermal sensitivity in schistosome infected mice, suggesting that this cytokine but not NGF plays a crucial role in schistosome-induced thermal hyperalgesia. Treatments with anti-inflammatory drugs support this hypothesis. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effects of IL-1 β and TNF- α on NGF levels in the knee joint in experimentally induced arthritis in adult mice. Out data showed that IL-1 β, but not TNF- α, induces an increase in NGF levels, while concomitant injection of both cytokines enhances the effect of IL-1 β on NGF presence. Analysis of NGF levels in the knee joint of carrageenan- and IL-1 β-induced inflammation after administration of antibodies against TNF- α supports this hypothesis. Our studies also showed that exogenous administration of NGF antibody reduces the enhanced level of TNF- α occurring in arthritic mice. This latter observation indirectly suggests that NGF is implicated in the upregulation of TNF- α in these animal models of joint inflammation. The functional significance of NGF or NGF antibody in inflammatory arthritis is discussed. [ABSTRACT FROM AUTHOR]

Numerous studies published in the last 10-15 years have shown that nerve growth factor (NGF), a polypeptide originally discovered in connection with its neurotrophic activity, also acts on cells of the immune system. NGF has been found in various immune organs including the spleen, lymph nodes, and thymus, and cells such as mast cells, eosinophils, and B and T cells. The circulating levels of NGF increase in inflammatory responses, in various autoimmune diseases, in parasitic infections, and in allergic diseases. Stress-related events both in animal models and in man also result in an increase of NGF, suggesting that this molecule is involved in neuroendocrine functions. The rapid release of NGF is part of an alerting signal in response to either psychologically stressful or anxiogenic conditions in response to homeostatic alteration. Thus, the inflammation and stress-induced increase in NGF might alone or in association with other biologic mediators induce the activation of immune cells during immunologic insults. A clearer understanding of the role of NGF in these events may be useful to identify the mechanisms implicated in certain neuroimmune and immune dysfunctions. [ABSTRACT FROM AUTHOR]

Our acute awareness of the cosmetic, psychosocial and sexual importance of subcutaneous adipose tissue contrasts dramatically with how poorly we have understood the biology of this massive, enigmatic, often ignored and much-abused skin compartment. Therefore, it is timely to recall the exciting, steadily growing, yet underappreciated body of evidence that subcutaneous adipocytes are so much more than just ‘fat guys’, hanging around passively to conspire, at most, against your desperate attempts to maintain ideal weight. Although the subcutis, quantitatively, tends to represent the dominant architectural component of human skin, conventional wisdom confines its biological key functions to those of energy storage, physical buffer, thermoregulation and thermoinsulation. However, already the distribution of human superficial adipose tissue, by itself, questions how justified the popular belief is that ‘skin fat’ (which actually may be more diverse than often assumed) serves primarily thermoinsulatory purposes. And although the metabolic complications of obesity are well appreciated, our understanding of how exactly subcutaneous adipocytes contribute to extracutaneous disease – and even influence important immune and brain functions! – is far from complete. The increasing insights recently won into subcutaneous adipose tissue as a cytokine depot that regulates innate immunity and cell growth exemplarily serve to illustrate the vast open research expanses that remain to be fully explored in the subcutis. The following public debate carries you from the evolutionary origins and the key functional purposes of adipose tissue, via adipose-derived stem cells and adipokines straight to the neuroendocrine, immunomodulatory and central nervous effects of signals that originate in the subcutis – perhaps, the most underestimated tissue of the human body. The editors are confident that, at the end, you shall agree: No basic scientist and no doctor with a serious interest in skin, and hardly anyone else in the life sciences, can afford to ignore the subcutaneous adipocyte – beyond its ample impact on beauty, benessence and body mass. [ABSTRACT FROM AUTHOR]

Rett syndrome is a severe neurological developmental disorder. In this syndrome, the high incidence of sudden death is correlated with an alteration of ventricular repolarization. The purpose of this study was to evaluate plasmatic levels of nerve growth factor (NGF) in Rett patients with prolonged corrected QT (QTc) interval in comparison with those of Rett patients with normal QTc. We observed 23 female Rett patients (9.9 ± 4.7 years). NGF plasma levels and QTc interval were measured in all patients. Student t-test was performed for statistical analysis. NGF plasma levels were significantly lower in Rett patients with QTc interval prolongation (QTc > 0.44 sec) in comparison with Rett patients with a normal QTc interval (4.5 ± 4.5 vs 11 ± 8.3 pg/ml, p = 0.02). The alteration of NGF levels, observed in Rett patients with a long QTc interval, may explain the presence of an altered ventricular repolarization associated with a higher risk of cardiac arrhythmias. [ABSTRACT FROM AUTHOR]

Nerve growth factor (NGF) is the first described neurotrophin that stimulates the growth and differentiation of nerve cells and promotes skin and peripheral tissue regeneration. Recent studies suggest that NGF influences endothelial cell proliferation and angiogenic activity. In view of these proposed regenerative effects, the efficacy of subcutaneous administration of highly purified murine NGF in a child with severe crush syndrome of the lower left limb has been evaluated. NGF 10 μg was administered subcutaneously every eight hours for seven days to the extensive ischaemic skin lesion of the calcaneal area. After treatment, gradual improvement of the ischaemic area was observed, while no side effects were noted.