bluuebunny/crossref_metadata_2025 huggingface dataset:
Internationalization and the Dynamics of Product Adaptation—An Empirical Investigation
Authors: Calantone Roger J. , Tamer Cavusgil S. , Schmidt Jeffrey B. , Shin Geon‐Cheol
Published: April 2004

Firms competing in foreign markets can choose to make no changes to the physical product and packaging, called a product standardization policy, which keeps costs low. The main drawback of such a policy is that the product might not satisfy customers. Conversely, firms may choose to modify, or to adapt, the physical characteristics or attributes of a product and its packaging to fit the needs and desires of consumers in different countries better, but this increases development, manufacturing, marketing, packaging, and distribution costs. Though product adaptation is a core aspect of customizing an export market offering, little research has investigated modifying the physical product and packaging. To be successful, an adapted product must add sufficient incremental revenue (through increased sales due to better satisfying customer needs and wants relative to competitive product offerings) such that the additional manufacturing and marketing costs that result from adapting the product are recovered.

In this article, a model of the product adaptation process is developed. Using mail surveys, information is gathered from managers in 239 U.S. organizations and 302 South Korean organizations, all of which export products. The goal was to understand better the motivation of firms to adapt their products for export markets as well as the performance implications of adapting products. Furthermore, the model was tested in these two countries to determine if the model is robust and to uncover differences between the United States and South Korea.

Using structural equation modeling to analyze the data, a positive association was found between the level of product adaptation and profitability at the project level. Second, U.S. firms appear to be more reactive when adapting products for export markets, doing so when laws and regulations in the export market mandate changes relative to the U.S. market. Conversely, South Korean firms appear to be more proactive and to adapt products even when not required by the governments of export markets. Third, greater international product adaptation is linked to a more responsive marketing organization with customer‐focused practices. Fourth, while a positive link was expected between business unit experience and the extent of international product adaptation, inconsistent results were found between the two country samples. For U.S. firms, it was found that greater experience in international business and product design capability is linked to a higher level of international product adaptation. For South Korean firms, however, a negative relationship was found. Greater international product adaptation occurred with less international business and product design experience. These findings are discussed, and areas for future research are noted.
Book Reviews
Authors: Smith Preston G.
Published: April 2004

Books reviewed in this issue:

Code Name Ginger: The Story behind Segway and Dean Kamen's Quest to Invent a New World

Open Innovation: The New Imperative for Creating and Profiting from Technology

Editor's note: The former section, “Books received for possible review in a future issue,” has been replaced by a more dynamic list available to reviewers. If you are interested in reviewing books for JPIM, please contact Preston Smith at preston@NewProductDynamics.com.
Screening for mutations in the lipopolysaccharide‐induced tumor necrosis factor‐alpha factor (litaf) gene in italian patients with charcot‐marie‐tooth disease.
Authors: Bellone E , Balestra P , Di Maria E , Pigullo S , Gulli R , Ajmar F , Mandich P
Published: April 2004

Mutations in LITAF (lipopolysaccharide‐induced tumor necrosis factor‐alpha factor), also referred to as SIMPLE, are associated with one of the dominantly inherited demyelinating forms of Charcot‐Marie‐Tooth disease (CMT1C). LITAF is a widely expressed gene, which maps on chromosome 16p13.1‐p12.3 and encodes a 161‐amino acid protein that may play a role in the degradation of proteins critical to peripheral nerve function. A cohort of 46 unrelated Italian patients affected with demyelinating peripheral neuropathy with dominant inheritance was screened for mutations in the LITAF gene. The three coding exons (2, 3 and 4) and flanking intron nucleotide sequence was examined by single strand conformation polymorphism (SSCP) and direct sequencing. All patients were negative for the 17p11.2 duplication and for mutations in the MPZ, PMP22 and EGR2 genes. SSCP analysis showed several electrophoretic variants in all exons. Direct sequencing demonstrated the presence of few single nucleotide substitutions. All of them were demonstrated to be common polymorphisms: one of them was non‐synonymous, two were synonymous and two were intronic. Based on the present analysis, LITAF mutations are not a frequent cause of autosomal dominant demyelinating neuropathies in our population.
Clinical history and new prognostic indicators in metachromatic leukodystrophy
Authors: Del Carro U , Biffi A , Baldoli C , Gerevini S , Amadio S , Fumagalli F , Roncarolo MG , Sessa M
Published: April 2004

Objective: To study clinical phenotypes and to increase knowledge of natural history of different variants of metachromatic leukodystrophy (MLD). Background: Little is known about factors influencing age of onset, progression rate and peripheral nerve involvement in MLD due to its rarity, heterogeneity and paucity of serial clinical and instrumental reports. Methods: 15 biochemically and molecularly characterized MLD patients were evaluated along a two‐year follow‐up period with clinical, electroneurographic (ENG) and brain MRI recordings. Results: Late infantile patients had a progressive and rapid course, whereas juvenile form showed marked variability. Different clinical presentations were associated with similar levels of ARSA activity; mutation screening indicated a high prevalence of rare or private mutations. In all late infantile and in the adult patient, ENG revealed a severe polyneuropathy. In juvenile patients a milder polyneuropathy or even normal tests were found. The earliest MRI change was periventricular white matter signal alterations, with initial involvement of posterior regions in a majority of late infantile patients, while in juvenile forms white matter lesions were mainly anterior. Conclusions: MLD course is highly variable and only partially influenced by age of onset, especially among juvenile patients. No clear‐cut correlations exist between clinical phenotype and biochemical or molecular characterization. The presence of peripheral neuropathy at onset seems a strong indicator of a poorer clinical outcome.
Dominant lamin A/C gene mutations can be associated with muscular dystrophy and peripheral neuropathy
Authors: Benedetti S , Previtali S , Toniolo D , Iannaccone S , Sferrazza B , Comi G , Carrera P , Ferrari M , Quattrini A , Bertini E
Published: April 2004

The coexistence of neurogenic and myogenic etiology in scapuloperoneal syndrome was not previously ascribed to a single gene. Defects in the lamin A/C gene (LMNA) have been associated with a variety of pathologies, affecting mainly muscular and adipose tissues, although recently reported also in autosomal recessive CMT2. We describe here two patients associating features of myopathy and neuropathy linked to dominant LMNA mutations. Patient 1 presented with distal weakness at lower limbs and elevated serum CK. Neurophysiological studies revealed a sensorimotor axonal neuropathy, whereas muscle biopsy showed signs of both neurogenic and myogenic features. Molecular analysis of LMNA gene revealed a heterozygous R571C substitution. Patient 2 presented with mild proximal weakness and fatigability. She was carrying a pacemaker for cardiac arrhythmia and had a family history of sudden death. Electronmyography showed mild myopathic changes. A muscle biopsy revealed chronic myogenic and neurogenic features including atrophic angular fibers and type grouping. Analysis of the LMNA gene showed a heterozygous four nucleotides deletion in exon 5 (864del4), leading to premature protein truncation. The same mutation was found in the asymptomatic younger brother. This report describes for the first time dominantly‐inherited alterations of the nuclear lamins affecting both muscle and peripheral nerve, and the clinical heterogeneity of LMNA mutations.
Standard deviation of the distances between consecutive epidermal nerve fibers as a parameter to study their distribution in human skin
Authors: Provitera V , Nolano M , Stancanelli A , Lullo F , Santoro L
Published: April 2004

Evaluation of epidermal nerve fiber (ENF) density by skin biopsy is nowadays considered as the gold standard for the diagnosis of small fiber neuropathies. ENFs show in normal subjects quite a regular distribution between the keratinocytes while in patients with small fiber neuropathies we can find, besides a loss of ENFs, evident anomalies of their distribution. Different patterns of nerve fiber dispersion can express different pathophysiological processes. The aim of this study was to define a tool capable of describing the distribution of ENF in the epidermis. For this purpose, we measured the distance between subsequent epidermal fibers on skin samples of healthy and diabetic subjects. We calculated the standard deviation of the interfiber length as a parameter (the dispersion index – DI) to evaluate the variability of the distance between consecutive ENFs. We selected thigh skin samples from five healthy subjects (age range 30–50 years) and five age and sex matched diabetic patients. Samples were processed using immunohistochemical techniques. Sections were PGP 9.5 and Collagen IV double stained to show nerve fibers and basement membrane. One randomly selected section for each subject was acquired for its entire length (usually seven 20x confocal images obtained from a z‐stack of 16 two micron optical sections) using a confocal microscope (CARV, Atto Biosciences, Rockville MD, USA). We measured the distance between consecutive ENFs along the entire epidermal length, using dedicated software (ScionImage, Scion Corporation Frederick, Maryland USA). In diabetic patients the DI resulted significantly higher than in normal subjects. We suggest that DI along with the ENF density can be useful to better characterize epidermal innervation.
Demyelinating motor guillain‐barré syndrome following rubella
Authors: Capasso M , Caporale CM , De Angelis MV , Cantarella C , Di Muzio A , Lugaresi A , Uncini A
Published: April 2004

A 20‐year‐old man developed weakness without sensory complaints ten days after rubella. Examination showed limb weakness and brisk tendon reflexes but no sensory abnormalities. Laboratory investigations revealed IgG and IgM anti‐Rubella and increased CSF protein content (0,8 g/L). Electrophysiological examination showed partial motor conduction blocks in eight nerves and normal sensory conductions even across the sites of CB. Brain and spinal cord MRI and SEPs were normal. The patient was treated with four plasmaphereses and fully recovered in six months. Conduction blocks gradually improved with increasing duration and abnormal temporal dispersion in proximal CMAPs. GBS has been rarely reported after rubella. Anti‐myelin basic protein antibodies have been found in a patient with a relapsing motor neuropathy following rubella vaccination. As antibodies cross‐reacted with a viral protein, molecular mimicry has been proposed as a pathophysiological mechanism. In our patient we did not find anti‐MBP antibodies and antibodies to‐glycolipids (GM1, GM2, GA1, GD1a, GD1b, GQ1b, sulfatides, galactocerebroside) were also negative. Indirect immunofluorescence after incubation of patient's serum on rabbit sciatic nerve and human sural nerve and roots was negative. Our patient confirms the occurrence of GBS following Rubella and shows some uncommon features: 1) hyperactive deep tendon reflexes; 2) demyelination selectively involving motor fibres; and 3) widespread early conduction blocks in intermediate nerve segments.
Acute demyelinating sensorimotor polyneuropathy in B‐cell lymphoma with IGM autoantibodies against glycolipid GD1B
Authors: Marfia GA , Pachatz C , Terracciano C , Leone G , Massa R
Published: April 2004

We report the case of an acute demyelinating sensorimotor polyneuropathy in a patient with IgM autoantibody against peripheral nerve myelin glycolipid GD1b produced by a B‐cell lymphoma. A 76‐year‐old woman presented with one month history of weight loss, paresthesias at the distal extremities of the four limbs followed by progressive and diffuse motor weakness. Neurological examination showed sensory ataxia, marked motor deficit (4 at MRC scale in proximal and distal muscle districts of limbs), superficial and proprioceptive sensory deficit with a stocking‐glove distribution, areflexia at lower limbs. Electroneurography documented absent sensory action potentials, prolonged distal latencies of motor action potentials, markedly slowed motor conduction velocities and evidence of conduction block and temporal dispersion of motor action potentials. Diagnosis of acute demyelinating sensorimotor polyneuropathy was established. CSF examination showed a slight increase of protein concentration. Haematological screening revealed a monoclonal gammopathy with high serum IgM (1151 mg/dL) and light chain kappa proteinuria. A total body CT scan evidenced a pelvic mass identified as low grade B‐cell lymphoma by rectal biopsy. Research for antiganglioside autoantibodies evidenced the presence of IgM antibodies against glycolipid GD1b. Autoantibodies anti‐Mag, anti‐Hu, anti‐Yo and anti‐Ri were negative. The pathogenetic role of serum autoantibodies against glycolipid GD1b in the development of demyelinization in the peripheral nervous system has already been documented either clinically in post‐infective polyneuropathies or experimentally in animal research. This is the first report of selective positivity of anti‐GD1b in polyneuropathy associated to B‐cell lymphoma supporting the possible paraneoplastic significance of these serum autoantibodies.
Thalidomide‐associated neuropathy in multiple myeloma
Authors: Morino S , Petrucci MT , Gragnani F , Ceschin V , Clemenzi A , Di Pasquale A , Antonimi G
Published: April 2004

Thalidomide is a neurotoxic immunomodulating agent currently used in Multiple Myeloma (MM). We prospectively evaluated the frequency and characteristics of peripheral neuropathy in a continuous series of 25 patients (13 M, 12 F; age 38–60, median 55 yrs) treated with thalidomide for MM. Patients underwent a neurological and neurophysiological evaluation before starting thalidomide therapy and monthly throughout duration of treatment. Sixteen patients (5 M, 11 F) developed neurophysiological characteristics of axonal sensitive damage and/or clinical peripheral neuropathy with distal sensory symptoms; treatment duration ranged between 95 and 572 days (median 298) in patients with neuropathy, and 49–264 days (median 162) in patients without neuropathy; the total amount of thalidomide taken ranged between 26 and 169 g (median 83 g) for patients with neuropathy and 13–170 g (median 51 g) for those without. In four patients, ENG alterations appeared before clinical symptoms, while in two patients they were not followed by clinical symptoms. In the remaining three patients, clinical symptoms preceded neurophysiological alterations. Age at onset of MM, disease duration before thalidomide therapy was started, total dose, duration of therapy and previous treatments were not correlated with neuropathy (multivariate logistic regression analysis). Female gender was a risk factor for developing neuropathy (OR 7.7).
CMTX: heterozygosity for a GJB1/CX32 mutation in a XXY male results in a mild phenotype
Authors: Milani M , Pareyson D , Taroni F
Published: April 2004

Mutations in the GJB1/Cx32 gene (Xq13.1) cause the most common X‐linked form of CMT (CMTX1) and are the most frequent cause of CMT disease after the CMT1A duplication. The disorder is characterized by a moderate‐to‐severe neuropathy in affected males and mild‐to‐no symptoms in carrier females. We report here a CMT1A‐negative family in which 4 females and 2 males were affected, exhibiting different disease severity. Molecular analysis of the GJB1/Cx32 gene uncovered a nonsense mutation (Arg22stop) in exon 2. The mutation, which had been previously described by others and observed by us in numerous other families, occurred in heterozygous form in the 4 females. However, while one of the two male patients was severely affected and shown to be hemizygous, as expected, the other was mildly affected and found to carry the mutation in heterozygous form. Genotyping at the SRY (Yp11.3) and DMD (Xp21) loci suggested the occurrence of the XXY genotype associated with Klinefelter syndrome. Microsatellite analysis indicated that the nondysjunctional error was of paternal origin, as it is usually observed in about half the cases. The patient had no children. At clinical examination, he exhibited a very mild neurologic phenotype and showed signs of hypogonadism (mild gynecomastia and small testes) as well as moderate cognitive impairment. Electrophysiologic, cytogenetic and endocrinologic investigations are in progress in order to define the unusual phenotype in this patient.
Molecular analysis of the litaf/simple and prx genes in patients with demyelinating charcot‐marie‐tooth (CMT) disease
Authors: Milani M , Cesari M , Baratta S , Caccia C , Balestrini MR , Riva D , Pareyson D , Taroni F
Published: April 2004

At least 20 genetic loci and 12 genes have been associated with the demyelinating form of CMT disease and related disorders. Mutations in the LITAF/SIMPLE gene (CMT1C; 16p13.3) have been recently suggested to be a relatively common cause of CMT1 after the CMT1A duplication and GJB1/Cx32 mutations (Saifi et al., ASHG 2003). We have screened for LITAF mutations a group of 120 CMT1 patients negative for CMT1A‐dup, MPZ, PMP22 and GJB1/Cx32. We have so far identified two relatively frequent polymorphisms and 3 severely affected patients who exhibited unique abnormal DHPLC profiles in ex2 and ex3. Unlike the dominant forms, only a limited number of loci have been linked to the rare recessive forms (AR‐CMT). The 4.9‐kb coding region of the periaxin gene (PRX, CMT4F) was scanned for mutations by DHPLC and sequence analysis in a group of 30 CMT1A/MPZ/PMP22/Cx32‐negative patients with severe demyelinating neuropathy. Three novel frameshift and nonsense mutations were identified in two AR families. Interestingly, in one family, two siblings exhibited different disease severity and were found to be compound heterozygotes for two mutations affecting the long (L‐periaxin) but not the short (S‐periaxin) variant of the protein, suggesting that S‐periaxin expression may account for clinical variability in these patients.
Influence of gender and pregnancy on CMT1A
Authors: Pareyson D , Dell’Anna E , DiMonda T , Scaioli V , Ciano C , Sghirlanzoni A , Lauria G , Morbin M , Laurà M , Solari A , Taroni F
Published: April 2004

Progesterone increases PMP22 expression. Transgenic rats overexpressing PMP22, a model of Charcot‐Marie‐Tooth type 1A (CMT1A), improve when treated with the progesterone antagonist, onapristone, and worsen with progesterone administration. CMT worsening during pregnancy (when progesterone secretion increases dramatically) has been occasionally reported. We investigated whether gender and pregnancy influence disease severity and course in CMT1A. We analyzed clinical and electrophysiological data of 94 CMT1A patients. Data in males and females were compared. We also determined disease severity in female patients in the fertile age as compared to males of the same age group. We retrospectively assessed influence of gestation on disease course in 20 CMT1A females through a comprehensive interview. There was no significant difference in disease severity in females as compared to males. Skeletal deformities, abnormalities of gait and deep tendon jerks, wasting, weakness, and sensory impairment were similar in the two genders. Nerve conduction abnormalities and decrease of sensory and motor evoked response amplitudes were comparable in the two groups. We found no difference in disease severity even when considering 15 to 50‐year‐old patients. Symptom worsening during pregnancy was reported by 4/20 CMT1A females. The effect of sex hormones on PMP22 does not result in a relevant difference in disease severity between the two genders. A few patients reported some worsening during pregnancy. Disease and normal controls need to be evaluated to establish whether this is a CMT1A‐related phenomenon.
Endothelin and endothelin‐converting‐enzyme‐1 in inflammatory neuropathies: an immunohistological study
Authors: Volpi N , Carbotti P , Greco G , Bibbò G , Alessandrini C , Giannini F
Published: April 2004

Endothelin (ET), a vasoconstrictive peptide, has a role in different biological processes. The isoform ET1, the most active in inflammation, is expressed by endothelial and glial cells, macrophages, leukocytes, fibroblasts and myocardiocytes. ET1 works as pro‐inflammatory cytokine in infectious vasculitis and myocarditis, and experimental myocardial infarction where antagonists of ET‐receptors are effective. Endothelin‐Converting‐Enzyme‐1 (ECE1), a zinc‐dependent metalloprotease, cleaves inactive precursors into mature ETs. ECE1 up‐regulation has been shown in experimental hypoglossal nerve axotomy and diabetic neuropathy. Interactions among cytokines, active mediators and matrix metalloproteases are currently being investigated in inflammatory neuropathies. We carried out immunohistochemical analysis of ET1 and ECE1 on sural nerves from nine patients with peripheral nerve vasculitis (NV), CIDP and CIDP‐diabetic neuropathy. NVs showed a clear ET1 up‐regulation on vessel walls of endoneurium and epineurium, notably in smooth muscle cells, and in inflammatory cells. ECE1 was expressed by endoneurial macrophages and Schwann cells. Both immunolocalizations were weaker in CIDP specimens, whereas CIDP‐diabetic neuropathy showed an intermediate pattern. Endothelin system may have a dual role in NV, as a vasoconstrictive agent contributing to fascicle ischaemia and as a pro‐inflammatory cytokine. Highest ECE1 expression in fascicles with severe axonal loss is possibly also due to its wider peptidase activity. In CIDP‐diabetic neuropathy, up‐regulation of ET1 and ECE1 is likely to depend on increase of ET‐receptors induced by hyperglycaemia.
Aquadynia. a manifestation of small fiber sensory neuropathy?
Authors: Brindani F , Gemignani F , Zinno L , Marbini A
Published: April 2004

Aquadynia is a rare phenomenon of water‐induced pain through presumed neural mechanisms. We describe two women in whom bathing was regularly followed by pain in the lower limbs, as a unique symptom (case 1) and, respectively, in the clinical context of an axonal polyneuropathy (case 2). Case 1: A 71‐year‐old woman complained, by age 69, of pruritus and pinprick‐like pain in the extremities that lasted about 20 minutes following bathing. Neurological examination and electroneurography were negative, as well as investigations for systemic diseases. Quantitative sensory testing (QST) showed abnormal cold‐pain sensation. Treatment with gabapentin was not effective. Case 2: A 69‐year‐old woman affected with HCV‐related cryoglobulinemia had numbness and aquadynia in the distal lower limbs and restless legs syndrome, in the last few months. Neurological examination showed absent ankle jerks, and decreased touch and vibration sense in the feet. Electroneurography demonstrated an axonal neuropathy. Aquadynia likely represents a manifestation of small fiber neuropathy. It is unclear whether it has to be viewed as a primary sensory phenomenon similar to allodynia, or a type of noradrenergic pain primarily due to autonomic dysfunction. Alteration of QST in case 1, and the association with obvious features of sensory neuropathy in case 2, may favour the sensory hypothesis.
Chronic ataxic neuropathy initially diagnosed as ataxic variant of guillain barré syndrome
Authors: Galassi G , Ruggeri A , Leone M , Cappelli G
Published: April 2004

Acute ataxic neuropathy is characterized by cerebellar‐like ataxia, insignificant weakness, areflexia, and mildly impaired sensation. Patients usually experience antecedent infections as in typical Guillain Barré syndrome. Clinical course is usually monophasic or relapsing with high titer of antiganglioside antibodies. A 40‐year‐old mildly hypertensive man was admitted because of acral paraesthesias, dysarthria, swallowing difficulties, and imbalance. On admission, the patient exhibited inconstant diplopia on lateral gaze, scanned speech, facial diplegia, cerebellar–like ataxia, tremor, areflexia, mild deep sensory loss, and nearly normal muscle weakness. Brain and spinal cord MRI was normal. Electrophysiology showed delayed F waves. Normal laboratory results included extensive hematologic tests, search for IgM paraprotein, antiganglioside antibodies, namely GT1a, GQ1b IgG, and cold agglutinins. CSF revealed albuminocytologic dissociation. A remote malignancy was excluded. Patient was treated with plasmaphoresis (PE) with transient benefit. Four weeks later, because of progression of cerebellar signs, the patient was newly admitted. Electrophysiology confirmed axonal and demyelinating features. CSF protein were 330 mg/dl (normal <45). PE, IVIG, methylprednisolone e.v. (1 g daily for 3 days, 500 mg for 2 days switched to oral regimen) and oral chlorambucil gave transient improvement. Nine months after first onset, the patient suffered new relapse. Renal biopsy was performed showing focal glomerular changes. In conclusion, the patient exhibited a chronic relapsing ataxic neuropathy without ophthalmoparesis or significant deep sensory disturbances, accompanied during acute phase by bifacial weakness in absence of antiganglioside antibody activity.
Clinical open‐label study for evaluation of efficacy and tolerability of oxcarbazepine in the treatment of neuropathic pain
Authors: Cafforio G , D’Avino C , Calabrese R , Siciliano G
Published: April 2004

Oxcarbazepine (OXC) is a keto‐10‐analogue of carbamazepine (CBZ) with linear pharmacokinetics and a better tolerability profile compared to CBZ. For this reason, OXC has been used in some studies in the treatment of neuropathic pain of various genesis, with administration doses ranging from 300 to 2400 mg/day. In this study we have assessed the efficacy and tolerability of OXC 600 mg/day, a dose considered, based on previous data, the minimal therapeutic dose in the treatment of neuropathic pain in some neuropathies, such as diabetic neuropathy. We have studied 11 patients affected by predominant sensitive painful neuropathy, of metabolic, compressive, inflammatory or dysimmune genesis, clinically characterized by paresthesias, dysesthesias and neuralgic pain not responding to other pharmacologic therapies. At the follow‐up analysis, 7 patients have referred an improvement of painful symptoms, as evidenced by Visual Analogue Scale score, the reduction of which ranged from 20 to 60% compared to baseline values. One patient interrupted the trial for incompliance, two patients for collateral events (excessive sleepiness) and another one for a worsening of clinical picture. In conclusion, our study suggests that OXC can be considered as a valid alternative in the treatment of painful neuropathies owing to its characteristics of tolerability and efficacy in such conditions.
RAT in vivo models of taxanes’ peripheral neurotoxicity following chronic intravenous administration
Authors: Canta A , Lanzani F , Galbiati S , Frigeni B , Giussani G , Marmiroli P , Tredici G , Traebert M , Müller L , Cavaletti G
Published: April 2004

The “taxanes” family includes some widely used antineoplastic agents, such as paclitaxel and docetaxel. Treatment with these microtubule‐stabilizing drugs is often associated with neurotoxicity, a potentially severe side effect limiting the clinical utility of these agents. To study the pathogenesis of taxanes’ neurotoxicity and to compare it to the effect of new agents, the availability of reliable in vivo models is warranted. In this study we developed chronic iv models for the assessment of “taxanes” peripheral neurotoxicity. Forty‐eight adult Wistar rats were divided into six groups of 8 animals each and treated as follows: paclitaxel or docetaxel at doses of 5, 10, 12.5 mg/kg 1q7d × 4 via a chronic jugular vein implant. The evaluation was based on the assessment of body weight and survival as a measure of general tolerability, and on the measurement of tail nerve conduction velocity, a neurophysiological method previously used in animal models of toxic peripheral neuropathies. The results were compared with those obtained in untreated or vehicle‐treated control rats. A clear dose‐dependent effect was evident both on general toxicity, and on neurophysiological changes measured at the end of the treatment (untreated controls = 41,9 m/sec, vehicle = 40,3 m/sec; paclitaxel 5 mg/kg = 32,5 m/sec, 10 mg/kg = 28,5 m/sec, 12.5 m/kg = 27,4 m/sec; docetaxel 5 mg/kg = 33,6 m/sec, 10 mg/kg = 27,8 m/sec, 12.5 mg/kg = 27,0 m/sec), demonstrating the usefulness of this new model system to investigate peripheral neurotoxicity mediated by taxanes, and potentially other drugs, under chronic treatment schedules.
Down‐regulation of receptors for semaphorins and VEGF in a transgenic model of CMT1A
Authors: Panico MB , Palumbo C , Vigo T , Nobbio L , Pisani V , Terracciano C , Schenone A , Modesti A , Massa R
Published: April 2004

Charcot‐Marie‐Tooth type 1 disease (CMT1) encompasses a group of primary demyelinating polyneuropathies caused by molecular defects of the Schwann cells (SC). Beside myelin damage, axonal atrophy and degeneration are invariably present and correlate with clinical severity. This is expected, since signals derived from SC regulate several axonal properties in myelinated fibers. However, the molecular mechanisms leading to axonal loss in CMT1 are still obscure. An important role in promoting axonal growth, guidance and fasciculation in peripheral nerve (PN) is played by semaphorins and the vascular endothelial growth factor (VEGF), by binding with specific receptors, the neuropilins (NP) and VEGF receptors (VEGFR). These molecules are expressed by peripheral neurons and SC, and induced in these cells during Wallerian degeneration. We have utilized a transgenic rat model of CMT1A to assess the immunoreactivity for NP and VEGFR in PN affected by PMP22 overdosage. These rats, in the hemizygote condition, show hypo/demyelination and axonal atrophy in PN. In the sciatic nerve of young hemizygotes we observed a substantial decrease of immunoreactivity for NP1 and VEGFR2, both in myelinated axons and SC, as compared to controls. Conversely, NP2 expression was slightly reduced. These data show that a down‐regulation of the semaphorin/neuropilin and VEGF signalling pathways may take place in CMT1. Such event may play a causative role in the axonal atrophy observed in this group of disorders.
Atypical rat cerebellar immunoreactivity in a patient with familial amyloid polyneuropathy
Authors: Mazzeo A , Laurà M , Giometto B , Ferlini A , Musumeci O , Autunno M , Vita G , Messina C
Published: April 2004

Objective: To describe an atypical immunoreactivity against Purkinje cells and glia of rat cerebellum in a case of transthyretin (TTR)‐related familial amyloid polyneuropathy (FAP). Background: TTR‐FAP is an autosomal dominant disease, usually presenting as a progressive sensory or sensorimotor polyneuropathy with autonomic disturbances. Case report: A 68‐year‐old man complained of a two‐year history of severe orthostatic hypotension, diarrhea and progressive sensorimotor polyneuropathy. The neurological examination showed a severe distal weakness at the four limbs, areflexia and distal impairment of pin‐prick sensation and proprioception. The patient was bed‐ridden because of the severe hypotension. Routine laboratory studies, autoantibodies, neoplastic markers and immunoelectrophoresis were negative; thyroid function was impaired with reduced levels of TSH, T3 and high level of anti‐tireoglobulin antibodies. An unusual reactivity against Purkinje cell membrane and glial cells of rat cerebellum white matter was found on immunohistochemistry. Western blot for the detection of onconeural antibodies (Hu, Ri, Yo and Amphiphysin) was negative. Electrophysiological studies showed a severe axonal sensorimotor polyneuropathy. Chest x‐ray and chest and abdomen CT scans did not reveal any malignancies. Genetic analysis revealed a Phe 64 Leu mutation on TTR gene. Discussion: The unusual immunoreactivity found in our patient has not been previously described. Our case report may be a warning to clinicians to suspect a genetic origin in patients with polyneuropathy and atypical pattern of neuronal immunoreactivity.
The spectrum of transthyretin gene mutations in italy: a report
Authors: Ravani A , Salvi F , Rimessi P , Lauria G , Vita G , Toscano A , Mazzeo A , Rappezzi C , Leone O , Tassinari CA , Calzolai E , Ferlini A
Published: April 2004

Transthyretin gene mutations causing autosomal dominant familial amyloidotic polyneuropathy (FAP) are often associated with cardiomyopathy. More than 80 amino acid variations have been described until now, showing a high allelic heterogeneity. We screened for TTR mutations 55 patients with mixed polyneuropathy with variable cardiac involvement. By means of direct sequencing of the full coding region we identified the following 26 genotypes: 2 Ser6 (polymorphism), 3 Met30, 4 Gln89, 2 Leu64 (TTT > TTG) and 7 Leu64 (TTT > CTT), 2 Ala49, 2 Leu68, 1 Thr34, 1 Arg50, 1 Glu53 and 1 Asn90. Interestingly, two Leu64 variants were due to a novel nucleotide variation TTT to TTG in codon 64 and 1 Asn90 mutation was found to co‐segregate with a FAP phenotype. Transthyretin mutations in hereditary amyloidosis deserve to be investigated in all cases of mixed polyneuropathy with cardiomyopathy both familial and sporadic. A marked allelic heterogeneity in Italy has been further confirmed by this study and PCR and direct sequencing seems to represent the more appropriate diagnostic tool for this pathology.
Peripheral neuropathy as initial sign of mitochondrial disorder
Authors: Manneschi L , Battisti C , Pesci I , Malandrini A , Santorelli FM , Scaglioni A , Federico A , Montanari E
Published: April 2004

Peripheral neuropathy is one of the clinical manifestations of mitochondrial disorders such as MELAS, MERRF, MNGIE, NARP, Leigh's syndrome, and Kearne‐Sayre syndrome. However, the prevalence of the neuropathy and its pathogenesis in mitochondrial diseases are not yet completely clarified. We describe the clinical case of a 40‐year‐old woman who, at 35 years of age, developed a progressive peripheral neuropathy that was the only and first clinical sign of disease for several years. In the family history a prevalence of severe cardiomyopathy in paternal lineage (the father and two brothers) is referred. Four years after the onset of the disease, our patient developed secondary thyroid hypothyroidism with multinodular gland and recently, bilateral cataract. Routinely laboratory investigations and neuroimaging study were normal. A mild increase of serum lactic acid was present (1.7 mmol/l, n.v. 0.3–1.3). Electrophysiological examination showed a sensorimotor neuropathy, with a prevalence of demyelinating pattern. Nerve biopsy confirmed myelin and axonal degeneration. Muscle biopsy showed 80% of negative cytochrome C oxidase fibres and mitochondria with paracrystalline inclusions. Biochemical analysis confirmed COX deficiency. Molecular genetic analysis excluded multiple deletions of mtDNA. The sequence of mitochondria DNA is being performed. In the literature there are a few cases of mitochondrial disorder with peripheral neuropathy at onset, usually juvenile cases. Our clinical case shows that a mitochondrial pathogenesis should be considered in the differential diagnosis of neuropathies.
Painful neuropathy vasculitis in HIV infection
Authors: Ferrari S , Cavallaro T , Lanzafame M , Malena M , Rizzuto N
Published: April 2004

The clinical picture of painful neuropathy is generally related to direct HIV involvement in the peripheral nervous system (PNS) or to neurotoxic effect of drugs contained in high activity antiretroviral therapy (HAART). We describe 2 patients with long‐standing HIV infection treated with HAART. Both patients developed a subacute distal painful symmetric neuropathy not influenced by analgesics and anti‐inflammatory drugs. Sural nerve biopsy revealed focal axonal loss, degeneration of myelinated fibers and epineurial necrotizing vasculitis with T‐cell infiltration of vessel wall. In both cases steroid therapy led to improvement of symptoms. Vasculitic neuropathy with a clinical picture of painful neuropathy is most unusual, the common clinical expression of vasculitis being mononeuritis multiplex. Our cases indicate that isolated vasculitis of peripheral nerve can occur in long‐standing HIV‐1 infection and the clinical presentation with distal symmetric polyneuropathy does not rule out such a diagnosis. High activity antiretroviral therapy is unable to prevent vasculitis in peripheral nerves; similar to a previous reported case of Guillain‐Barré syndrome, an immune reconstitution phenomenon plays a role in the induction of a vasculitic autoimmune process. In conclusion, in cases of DSPN with intense and persistent symptoms, a nerve biopsy is necessary for a differential diagnosis to be made and an appropriate and effective therapy given.
Are giant axons a pathological marker of charcot‐marie‐tooth neuropathy type 2E?
Authors: Cavallaro T , Fabrizi GM , Angiari C , Lus G , Cabrini I , Cotrufo R , Rizzuto N
Published: April 2004

Background: According to electrophysiological and pathological criteria Charcot Marie Tooth (CMT) disease includes primary demyelinating forms (CMT1) and neuropathies with primary axonal loss (CMT2). In CMT1, genetic analysis provided some associations between characteristic lesions and different proteins. In CMT2, four genes were identified recently (CMT2A, B, D, E); the molecular diagnosis is complex and phenotypical hallmarks are lacking. Objectives: To describe the nerve biopsy in three pedigrees with CMT2E caused by mutations of the neurofilament‐light chain gene (NF‐L): two pedigrees from Campania sharing a Pro22Ser substitution in the head domain of protein and one pedigree from Apulia with a novel Leu268Prol substitution in the central rod domain. In all three pedigrees electrophysiology was consistent with a mixed, demyelinating and axonal neuropathy. Results: The three patients analysed revealed a primary axonopathy characterized by giant axonal swelling filled with densely packed neurofilaments and some atrophic axons. Conclusions: We propose that, in the diagnostic work up of CMT2, giant axons may orientate towards CMT2E. The pathological alterations detected correlate intuitively with an altered function of the neurofilaments which constitute the axonal cytoskeleton and are critical for radial growth and for axonal transport.
A splice‐junction mutation in SBF2 gene causes autosomal recessive charcot‐marie‐tooth disease (CMT4B2) in a family from southern italy
Authors: Conforti FL , Muglia M , Mazzei R , Valentino P , Patitucci A , Bono F , Magariello A , Sprovieri T , Senderek J , Bergmann C , Nisticò R , Gabriele AL , Peluso G , Quattrone A
Published: April 2004

Autosomal recessive Charcot‐Marie‐Tooth disease type 4 (CMT4) comprises a group of clinically and genetically heterogeneous disorders of the peripheral nervous system. At least 10 loci are responsible for autosomal recessive CMT and six genes have been identified so far. In this study, we report a small pedigree with a recessive form of CMT (CMT4B) from Southern Italy. There were six individuals in two generations with two affected subjects. We performed haplotype analysis using highly polymorphic microsatellite markers located on chromosome 11p15. Subsequently, the coding region of the Sbf2 gene was sequenced by using primers flanking intron‐exon boundaries. Mutational screening of Sbf2 revealed a homozygous mutation in the splice‐junction donor‐acceptor site of intron 32 (+1G→C) in the affected patients. The variation was also confirmed by digestion with restriction enzyme Alu I and it was absent in 100 control chromosomes examined. This is the first finding of a mutation in the Sbf2 gene that alters the correct splicing of the gene. Furthermore, these data confirm that mutations in the Sbf2 gene are causative of CMT4B2.
Narrowing of the critical region in autosomal recessive spastic paraplegia linked to the SPG5 locus
Authors: Muglia M , Criscuolo C , Magariello A , De Michele G , Scarano V , D’Adamo P , Ambrosio G , Gabriele AL , Patitucci A , Mazzei R , Conforti FL , Sprovieri T , Morgante L , Epifanio A , La Spina P , Valentino P , Gasparini P , Filla A , Quattrone A
Published: April 2004

Hereditary spastic paraplegias are neurodegenerative disorders characterized clinically by progressive spasticity of the lower limbs; they are inherited as autosomal dominant, autosomal recessive and X‐linked traits. We have analyzed four autosomal recessive HSP families gathered from southern Italy. We performed genetic analysis using microsatellite markers associated with SPG5, SPG7, SPG11 and SPG14. Positive lod scores were obtained with markers located on chromosome 8. The lod scores for the four combined families were significantly higher than 3 for D8S509, D8S1102, D8S1723 and D8S260 with a maximum two‐point lod score at θ = 0 of 3.99 for the marker D8S260. In one of the examined families, the haplotype analysis suggests two key recombination events demonstrating that the gene is localized in the 11 cM region flanked by markers D8S285 and D8S544, refining the ARHSP region by approximately 22 cm. We also analyzed five candidate genes localized within the HSP region: TOX, syndecan‐binding‐protein (SDCBP), RAB2, CA8 and PENK, but we did not find disease causing mutations.
Unravelling the molecular basis of CMT4B pathology
Authors: Bolis A , Previtali S , Bussini S , Dina G , Dati G , Feltri ML , Quattrini A , Wrabetz L , Bolino A
Published: April 2004

Charcot‐Marie‐Tooth type 4B (CMT4B) disease is a severe autosomal recessive peripheral neuropathy with childhood onset, characterised by progressive muscular atrophy and weakness in the distal extremities, sensory loss, severely decreased nerve conduction velocities, and demyelination with myelin outfoldings in the peripheral nerve. We demonstrated that CMT4B is caused by loss of function mutations in the Myotubularin‐related 2 gene, MTMR2, on chromosome 11q22 (Bolino et al., Nat Genet 25:17–19, 2000). MTMR2 belongs to the myotubularin family of protein phosphatases, of which, myotubularin (MTM), mutated in the X‐linked myotubular myopathy (XLMTM) is the founder member. MTMR2 shows specific activity towards phosphatidylinositol 3‐phosphate and 3,5‐biphosphate, PI(3)P and PI(3,5)P2, respectively. However, how abrogation of this lipid phosphatase activity is leading to the specific disease phenotype has not yet been demonstrated. To elucidate the biological role of MTMR2 in the nerve, we performed an extensive expression analysis of this protein in the peripheral nervous system. Since MTMR2 was demonstrated to be ubiquitously expressed also within the nerve, we sought nerve‐specific interactors using the yeast two‐hybrid approach. The neurofilament light chain protein, NF‐L, mutated in various CMTs including axonal type CMT2E, and demyelinating Dejerine‐Sottas syndrome, was found to interact with MTMR2 in Schwann cells as well as in neurons. Since NF‐L is specifically expressed in the nervous system, the interaction between MTMR2 and NF‐L would explain why loss of a ubiquitously expressed phosphatase affects specifically the nerve (Previtali and Bolino, Hum Mol Genet 12:1713–1723, 2003). To model the CMT4B pathology, we generated a general knock‐out mouse arising from inactivation f Mtmr2 in all cells. The characterisation of this animal model is underway. Overall, Mtmr2 null mice display a milder phenotype with respect to the human disorder. The morphological analysis of the peripheral nerve of this mouse line performed at P28 revealed the presence of myelin outfoldings, which are the hallmark of CMT4B pathology.
Intraepidermal innervation and tail nerve conduction velocity in neurotoxicity models: results of a correlation study in normal and pathological conditions
Authors: Borgna M , Lombardi R , Lauria G , Grezzi P , Savino C , Bianchi R , Oggioni N , Canta A , Lanzani F , Galbiati S , Frigeni B , Giussani G , Tredici G , Cavaletti G
Published: April 2004

Animal models of human diseases affecting the peripheral nervous system are widely used to assess the pathogenesis of neurotoxicity and to compare the effect of new agents. Several behavioural, pathological and neurophysiological methods have been used, and each has advantages and disadvantages. A major goal in the study of neurotoxicity would be to assess the damage in the same way in animal models and in humans. In this study we correlated the neurophysiological results obtained in normal rats and in rats treated with cisplatin 2 mg/kg q3d × 8 with the density of intraepidermal fibers (IEF) obtained in skin biopsy specimens. The aim was to investigate the possible role of a minimally invasive procedure such as skin biopsy as an alternative method to assess the peripheral neurotoxicity of antineoplastic drugs. The nerve conduction velocity (NCV) in the tail nerve was assessed in thirty‐six young adult female Wistar rats which were left untreated, or treated with erythropoietin (EPO), cisplatin (CDDP) or EPO + CDDP. CDDP and CDDP + EPO‐treated rats had a significantly reduced NCV vs. age‐matched untreated rats. At sacrifice, skin specimens were obtained. The density of IEF was calculated by 2 independent blinded examiners and the correlation existing between NCV and IEF was highly significant (r = 0.670, p < 0.001). This preliminary result suggests that IEF should be evaluated in other animal models and might represent a useful tool to study peripheral neurotoxicity also in humans.
Increase of pro‐inflammatory cytokines in chronic sensory ataxic neuropathies
Authors: Sanvito L , Santuccio G , Speciale L , Sgandurra M , Merlino L , Ceresa L , Ferrante P , Nemni R
Published: April 2004

Chronic sensory ataxic neuropathies (CSAN) are characterised by a predominant involvement of sensory large fibers. We previously demonstrated the presence of serum antibodies against neuropathy‐related antigens in about 50% of these patients. The aim of our study was to analyse peripheral T and B cell subsets and cytokine network in CSAN patients in order to evaluate their immunological profile. We studied 10 CSAN patients. ELISA test detected the presence of neuropathy‐related autoantibodies in 5 out of 10 patients. We evaluated, by flow cytometry, CD3+, CD3 + DRII+, CD4+, CD8+, CD16+ and CD19+ cells subset distribution, and intracellular IL2, INFg, IL1, IL6, IL10 and TNFa production by CD4+, CD8+ and CD14+ cells. Results were compared with healthy controls (HC); statistical analysis was performed. Compared to HC, CSAN patients showed no significant difference in lymphocyte subset distribution. We found a significant increase of IL6 in CD4+, CD8+ and CD14+ cells. INFg and TNFa were increased in CD4+ cells. TNFa was decreased in CD14+. Our results show an increase of pro‐inflammatory cytokines not associated with imbalance of T and B cells subsets. These data support an activation of the immune system in CSAN, as previously suggested by the detection of neuropathy‐related autoantibodies.