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Carotenoid cleavage dioxygenases (CCDs) are a class of enzymes involved in the biosynthesis of a broad diversity of secondary metabolites known as apocarotenoids. In plants, CCDs are part of a genetic family with members which cleave specific double bonds of carotenoid molecules. CCDs are involved in the production of diverse and important metabolites such as vitamin A and abscisic acid (ABA). Bixa orellana L. is the main source of the natural pigment annatto or bixin, an apocarotenoid accumulated in large quantities in its seeds. Bixin biosynthesis has been studied and the involvement of a CCD has been confirmed in vitro. However, the CCD genes involved in the biosynthesis of the wide variety of apocarotenoids found in this plant have not been well documented. In this study, a new CCD1 gene member (BoCCD1) was identified and its expression was charaterized in different plant tissues of B. orellana plantlets and adult plants. The BoCCD1 sequence showed high homology with plant CCD1s involved mainly in the cleavage of carotenoids in several sites to generate multiple apocarotenoid products. Here, the expression profiles of the BoCCD1 gene were analysed and discussed in relation to total carotenoids and other important apocarotenoids such as bixin.

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A reliable protocol is described for isolation of large full-length cDNA from Bixa orellana mature tissues containing large quantities of pigments, phenols, and polysaccharides. This protocol involves the optimization of a commercial RNA extraction protocol in combination with a long distance reverse transcript PCR protocol. The principal advantages of this protocol are its high RNA yield and quality. The resulting RNA is suitable for RNA expression evaluation and production of large, full-length cDNA. This is the first time RNA has been isolated from all mature tissues in the tropical perennial plant B. orellana and has been proved viable for downstream applications, especially important for molecular biology studies on this economically important pigment-producing plant.

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Dysfunction of small fibers may appear in isolation or associated with large fiber lesions. In some acute neuropathies, such as pandysautonomia, small-fiber impairment is relatively pure but it may also appear in disorders with prominent somatic damage, such as Guillain-Barré syndrome, in which autonomic failure worsens the prognosis. At the present time, chronic idiopathic distal small-fiber neuropathy is diagnosed more frequently, and in some prevalent disorders, such as diabetic or amyloidotic polyneuropathies, small-fiber dysfunction is very noticeable. In pure autonomic failure, a peripheral autonomic failure exists, distinguishing it from multiple-system atrophy. Complex regional pain syndrome is a severe condition in which small fibers are responsible for disabling signs and symptoms, and only instrumental recordings lead to the proper treatment. Standard neurophysiological techniques evaluate large myelinated fibers exclusively. Small-fiber polyneuropathy has been considered as a type of somatic neuropathy, but thin myelinated and unmyelinated fibers are responsible not only for temperature and pain perception but also autonomic function. For instance, full autonomic evaluation is needed in some clinical situations such as autonomic failure in the elderly or orthostatic intolerance syndrome. To evaluate small-fiber impairment we need a battery of sensitive, reproducible, specific and noninvasive tests covering somatic and autonomic systems. In this review, we describe and analyze a number of neurophysiological techniques used to diagnose and characterize small-fiber dysfunction in humans. These include cardiovascular monitoring, sudomotor testing, pupillary responses and quantitative sensory tests, and also to some extent thermography and laser evoked potentials. The use of such techniques has proven useful not only for diagnosis, but also to guide adequate therapy and optimize follow-up.

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INTRODUCTION: Standard neurophysiological techniques evaluate thick myelinated fibers. Yet, peripheral nerves are equally composed of thin myelinated and unmyelinated fibers. The latter are responsible for autonomic function as well as temperature and pain perception. DEVELOPMENT: Microneurographic studies are restricted to investigation laboratories. Since the techniques are complex and invasive, their performance is still poor for clinical purposes and some of the components to be analyzed, such as cardiovagal, cannot be directly recorded. The clinical need to evaluate the functions regulated by the autonomic nervous system (ANS) had led to devising a series of tests which, in most cases, rely on reflex responses evoked by already known standardize stimuli. The battery chosen has to be non invasive, reproducible, specific, providing relevant data to the investigated function, with a readily available technology, which has to be managed being aware of the physiological and pathological factors that might bear an influence on the results. The recent development of heart rate and blood pressure power spectral analysis, provides a new interesting insight for quantification of ANS abnormalities. The study of thermography and thermometry of body surface brings forward evidence on the activity of other thin and unmyelinated fibers components of the peripheral nerve spectrum. CONCLUSION: The adequate management of the above mentioned tests gives rise to a more extensive and appropriate knowledge of the whole peripheral nerve fiber spectrum.

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INTRODUCTION: Disfunction of thin myelinated and unmyelinated fibers may appear isolated or in association with large-myelinated fibers lesion. Small-fiber neuropathy includes autonomic and sensory symptoms, most prominent of them thermo-algesic deficits. DEVELOPMENT AND CONCLUSION: In some acute neuropathies, small-fiber lesion is relatively pure, as in pandysautonomia, but it also appears in disorders with prominent somatic involvement, such as the Guillain-Barre syndrome, in which case autonomic symptoms worsens the prognosis. Small-fiber dysfunction is important in certain diseases that involve different components of the nervous system, like paraneoplastic syndromes and porphyria. Some drugs and toxic substances may damage thin myelinated and unmyelinated fibers. Nowadays, chronic idiopathic small-fiber neuropathy is diagnosed more frequently, because of the recent development of techniques that selectively evaluate this peripheral nerve component. Hereditary sensory and autonomic neuropathies can also be studied. Small-fiber dysfunction is very prominent in some diseases, e.g. diabetes mellitus and amyloidosis. In the pure autonomic failure, only the peripheral component of the autonomic nervous system is affected, and this feature is the key to make diagnosis versus multisystem atrophy. There are situations in which there is no clear deviation from normality, namely old age autonomic failure and orthostatic intolerance syndrome in which autonomic evaluation is mandatory.

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INTRODUCTION: Ambulatory monitoring of EEG by means of a cassette recording (A/EEG) allows cerebral electrical activity or other biological signals to be recorded during long periods, not less than 24 hours, with the patient being free to move around in his usual surroundings or in the hospital. MATERIAL AND METHODS: The recordings are made using a system of electrodes, recording apparatus and software. CONCLUSIONS: The A/EEG is useful in the study of various diagnostic procedures, basically in the diagnosis and characterization of epilepsy. It is also useful in presurgical evaluation of patients with drug-resistant epilepsy in whom surgical treatment is being considered. In these cases, the possibility of prolonged monitoring allows some of the patient's crises to be recorded, which is very useful in finding the site of the lesion. It is also useful when studying patients with hypersomnia or disorders of the wakefulness-sleep cycle. It is complementary to multiple sleep latency tests (TLSM) and polygraphic recordings of nocturnal sleep, since it allows the quantity, duration and type of sleep to be recorded during prolonged monitoring for more than 24 hours.

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INTRODUCTION: Electroencephalography, together with new neuroimaging techniques is still the main diagnostic tool for pre-operative assessment of epilepsy. The electrical episodes, both ictal and intercritical, are random paroxystic phenomena with regard to presentation, so the conventional EEG is sometimes of limited value. Therefore, in recent years, long duration EEG monitoring techniques have been favoured, with or without simultaneous recording of clinical behaviour, permitting recording for an indefinite period. Amongst these techniques, monitoring using cassette recording (A/EEG) if of specific interest since it permits the study of outpatients. OBJECTIVE: The main aim of surgical treatment of epilepsy is to abolish or reduce the number of crises. Therefore it is essential to localize the epileptogenic area as precisely as possible. The A/EEG has made a major contribution to the procedure of pre-operative assessment because of the considerable information it gives regarding the recording and character of the crises. MATERIAL AND METHODS: During the past three years, preoperative assessment has been carried out in our hospital on 44 patients with drug-resistant partial epilepsy. A total of 103 long duration cassette recordings (A/EEG) were done. Of these, 58 were done with surface electrodes and individualized set-up depending on the EEG findings, 8 with semi-invasive (sphenoid) electrodes, and 37 recordings used implanted sub-dural electrode strips. RESULTS: Combined study of the results obtained with these techniques permitted localization of the lesive-epileptogenic complex in 42 of these patients. We present the results obtained and conclude that A/EEG monitoring is useful in the pre-operative assessment of epilepsy.

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The potential differences in hematologic profiles of blood samples drawn simultaneously from the right utero-ovarian vein and from the upper extremity were investigated in four patients with uncomplicated molar pregnancy in stable obstetric conditions. The patients had undergone no previous chemotherapy and were scheduled for total abdominal hysterectomies. The dominant abnormalities in uterine venous blood were prolongation of thrombin time; shortening of activated partial thromboplastin time; positive protamine sulfate test; and increase in coagulation factors II and VII, with a tendency to low values in factor V. Peripheral samples gave almost parallel results in all altered and normal tests, except in one case with very striking differences in factors II, V, VII and X. Several local and systemic influences are discussed. It is concluded that molar pregnancy seems to have important systemic mechanisms affecting the stability of the blood coagulation homeostasis, which act in addition to those at a local level.

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