Docosahexaenoic acid elevates trans-18:1 isomers but is not directly converted into trans-18:1 isomers in ruminal batch cultures.

Pathways of docosahexaenoic (DHA) biohydrogenation are not known; however, DHA is metabolized by ruminal microorganisms. The addition of DHA to the rumen alters the fatty acid profile of the rumen and milk and leads to increased trans-18:1 isomers, particularly trans-11 18:1. This study included 2 in vitro experiments to identify if the increase in trans-11 C18:1 was due to DHA being converted into trans-11 18:1 or if DHA stimulated trans-11 products from biohydrogenation of other fatty acids. In each experiment, ruminal microorganisms collected from a lactating Holstein cow were incubated in 10-mL batch cultures for 0, 6, 24, and 48 h and a uniformly (13)C-labeled DHA was added to the cultures at 0 h as a metabolic tracer. Experiment 1 tested 0.5% DHA supplementation and experiment 2 examined 1, 2, and 3% DHA supplementation to determine if the level of DHA effected its conversion into trans-11 18:1. In both experiments, any fatty acid that was enriched with the (13)C label was determined to arise from DHA. Palmitic (C16:0), stearic (C18:0), all trans-18:1, eicosanoic (C20:0), and docosanoic (C22:0) acids were examined for enrichment. In experiment 1, the amount of trans-18:1 isomers increased 0.415 mg from 0 to 48 h; however, no label was found in trans-18:1 at any time. Docosanoic acid was highly enriched at 24h and 48 h to 20.2 and 16.3%. Low levels of enrichment were found in palmitic and stearic acids. In experiment 2, trans-18:1 isomers increased 185, 256, and 272% from 0 to 48 h when DHA was supplemented at 1, 2, and 3%, respectively; however, as in experiment 1, no enrichment occurred of any trans-18:1 isomer. In experiment 2, low levels of label were found in palmitic and stearic acids. Enrichment of docosanoic acid decreased linearly with increased DHA supplementation. These studies showed that trans-18:1 fatty acids are not produced from DHA, supporting that DHA elevates trans-18:1 by modifying biohydrogenation pathways of other polyunsaturated fatty acids.

[Tumour-like lesions of the orbit - clinical symptoms, difficulties in differentiation and options for therapy]. / Klinische Symptomatik, differenzialdiagnostische Schwierigkeiten und Therapieoptionen bei tumorartigen Läsionen der Orbita.

Tumour-like lesions are very rare entity among orbital diseases. Between 1996 and 2007, 83 patients with orbital disorders were treated and three of them complained about different tumour-like lesions: giant cell granuloma, eosinophilic granuloma and fibrous dysplasia. Because of the considerable increase of functional disorders and tumour growth, all of the patients underwent surgical treatment. In this paper we describe the clinical symptoms, diagnostics and the methods of therapy for each of the lesions. The results shows that the preoperative neuro-ophthalmological examination as well as radiological imaging are necessary to assess the motility, visual field, vision, eye ball dislocation and to plan the surgical strategy. In terms of this descriptive character of the paper, we hope to submit some relevant information in order to improve the diagnostics and therapeutic procedures for tumour-like lesions of the orbit.

[Intraorbital tumours of the peripheral nervous system]. / Intraorbitale Tumoren des peripheren Nervensystems.

In neoplastic processes in the orbital area, a not significant part of these new entities is caused by tumours of the peripheral nervous system. Due to a clinically similar behaviour and radiological features also being similar, these tumours sometimes cause differential diagnostic difficulties. Our experience with these tumours shows that they do not have a uniform symptom complex. Exophthalmus and displacement of the globe are the most visible symptoms besides pain, motility disturbances, reduced visus and double images. Clinical imaging allows the exact localisation of the tumour and makes it easier to choose the best surgical method. However, imaging does not allow the exact assessment of the entity or enable a decision to be made about the possible malignancy of the tumour. Therefore, a representative biopsy and a histopathological examination are essential. Then, a complete tumour resection should be performed using the most careful incision possible to avoid postoperative complications.

Probable adult polyglucosan body disease.

Adult polyglucosan body disease is a clinicopathologic entity characterized by progressive upper and lower motor neuron dysfunction, sensory loss in the lower extremities, sphincter dysfunction, and occasionally dementia. Pathologically, numerous large polyglucosan bodies are noted in peripheral nerves, cerebral hemispheres, and the spinal cord, as well as in other systemic tissues. We present a case of probable adult polyglucosan body disease based on clinical history and examination, magnetic resonance images, and sural nerve biopsy findings.

Calibration of methods for determining numbers of dorsal root ganglion cells.

Numbers of primary afferent neurons underlie important generalizations concerning the organization of primary sensory systems. A major difficulty, however, is that different investigators do not agree on the neuronal counts. The problem, in our opinion, is that the various methods used to determine these numbers do not provide the same results. Thus to be certain that a method provides accurate counts, calibration is necessary. To do this, true numbers of ganglion cells were determined by serially reconstructing significant parts of four rat lumbar dorsal root ganglion cell populations. Then 6 commonly used methods of counting neurons were used to determine neuron numbers for these same populations. The data indicate that the empirical method, using the modifications recommended in this paper, estimates numbers of neurons with the needed accuracy whereas the other 5 do not. Thus, of the tested counting procedures, the empirical method is recommended. If other methods are to be used, they should also be calibrated.

The receptive part of the primary afferent axon is most vulnerable to systemic capsaicin in adult rats.

The present study shows that systemic capsaicin in adult rats results in a significant loss of axons in the subepidermal nerve plexus of the posterior leg but no loss of axons in the sural nerves of these same animals. These data are interpreted as indicating that the receptive part of the peripheral sensory axon is destroyed but that the cell body and most of the peripheral axon remains intact. Thus we suggest that the receptive part of the peripheral sensory axon is the most vulnerable part of the primary afferent neuron to capsaicin in these animals. These findings may explain the observation that adult rats treated with systemic capsaicin are deficient in their responses to certain painful stimuli but usually do not show obvious signs of primary afferent neuron death. We also suggest that as the dose of capsaicin is increased the whole neuron dies. It remains to be determined if the peripheral damage reported here is related to the striking loss of primary afferent markers in the dorsal horn that is also seen after this treatment.

Unmyelinated primary afferent fiber stimulation depletes dynorphin A (1-8) immunoreactivity in rat ventral horn.

The present study demonstrates many dynorphin (DYN)-immunoreactive fibers and presumed presynaptic terminals in rat lumbar ventral horn. The fibers and terminals seem to arise largely from DYN-containing intrinsic neurons in the dorsal horn. The majority of the presumed terminals closely surround a subpopulation of motoneurons that tend to be located in flexor motoneuron columns. Acute C fiber, but not A fiber, primary afferent stimulation depletes the ventral horn DYN immunostaining. We interpret these findings to indicate that the spinal DYN neurons are well positioned to serve both as modulators of nociceptive input and as interneurons in motor reflexes. We further hypothesize that the depletion of DYN-immunoreactivity that follows either acute C fiber stimulation or intense nociceptive stimuli may be the trigger for the upregulation in spinal cord DYN that occurs in models of chronic pain states.

Percentages of dorsal root axons immunoreactive for galanin are higher than those immunoreactive for calcitonin gene-related peptide in the rat.

The present study shows that 28% of the myelinated and 27% of the unmyelinated axons in the L5 and S1 rat dorsal roots are immunolabeled for galanin. By contrast only 10% of the myelinated and 15% of the unmyelinated axons are immunolabeled for calcitonin gene-related peptide, which is the numerically predominant primary afferent peptide marker for dorsal root ganglion cells. Thus galanin, because of its presence in so many primary afferent fibers, emerges as an important primary afferent marker. In addition, since our data also show that galanin is present predominantly in unmyelinated and fine myelinated sensory axons, a hypothesis is that it is particularly concerned with the transmission of noxious information.

Direct electrical stimulation of the superior ovarian nerve in rats causes an increase in neuronal activity in the ipsilateral ovarian plexus nerve.

Ovarian blood flow and neuronal activity in the ovarian plexus nerve (OPN) were monitored before, during and after direct electrical stimulation of the central cut end of the ipsilateral superior ovarian nerve (SON). Blood flow did not change during the observation period. Stimulation of the SON increased the frequency of action potentials in the OPN, suggesting an ovarian reflex pathway mediated by the SON and the OPN.

Do neuropeptides in the dorsal horn change if the dorsal root ganglion cell death that normally accompanies peripheral nerve transection is prevented?

Peripheral nerve section causes the death of dorsal root ganglion cells and changes in neuroactive peptides in the dorsal horn of the spinal cord. The relationship between these 2 events has not been previously studied, however. One approach would be to prevent sensory cell death and then determine changes in peptide immunoreactivity. To do this, transected rat sciatic nerve stumps were placed in an impermeable silicone tube for one month. The tube was then removed and after 30 additional days the cells were counted. The data indicate that no cell death occurred. We conclude that the sensory cells are first saved due to some factor present in the tube, and then after 30 days, the cells become independent of the tube and its contents. In these same animals, all of the peptides we examined were significantly changed. Four of the peptides, calcitonin gene-related peptide (CGRP), substance P (SP), cholecystokinin octapeptide (CCK) and galanin (GAL) were significantly depleted in the medial L4-L5 superficial dorsal horn, and vasoactive intestinal polypeptide (VIP) was significantly increased. We conclude that there are major changes in spinal peptide systems following peripheral nerve transection even if there is no accompanying death of sensory neurons. Thus we suggest that dramatic central changes in peptide immunoreactivity following peripheral nerve transection are independent of the sensory cell death that usually occurs in response to this injury.

The effects of A- and C-fiber stimulation on patterns of neuropeptide immunostaining in the rat superficial dorsal horn.

The present study determines the effects of sciatic nerve stimulation at intensities that activate A-fibers alone or both A- and C-fibers on immunostaining for substance P (SP), cholecystokinin-octapeptide (CCK-8), galanin (GAL), dynorphin (DYN) and vasoactive intestinal polypeptide (VIP) in the superficial dorsal horn of the rat spinal cord. The goal of this study is to provide a more precise spatial localization of the sites of release or accumulation of these compounds in relation to specific types of stimuli. Following A-fiber stimulation, there was no significant change in immunostaining for any of these compounds. However, A- and C-fiber stimulation resulted in major changes. For SP, CCK-8, GAL and DYN there was a large and significant loss of immunostaining in medial regions of the dorsal horn. This is the area where sciatic nerve primary afferent fibers terminate and the depletion is probably correlated with activity in these fibers. By contrast, VIP immunostaining is increased in the lateral part of the superficial cord, which is outside of the central sciatic afferent fiber terminations. This indicates that the increase is not in the fine sciatic sensory axons that are directly stimulated. As a final point, the fact that C-fiber but not A-fiber stimulation causes marked changes in the immunocytochemical distribution of all these compounds is further evidence, albeit indirect, that they are involved in nociceptive information processing.

Anatomical localization of afferent and postganglionic sympathetic neurons innervating the rat ovary.

The fluorescent retrograde tracer, True blue, was applied directly to either the superior ovarian nerve (SON) or the ovarian plexus nerve (OPN) in the rat. Afferent perikarya were located in lower thoracic-upper lumbar dorsal root ganglia and projected to the ovary via both nerve routes. Postganglionic sympathetic efferent perikarya were located in both prevertebral and thoracolumbar paravertebral ganglia and also utilized both the SON and OPN to reach the ovary. The significance of the dual origin of postganglionic sympathetic neurons innervating the rat ovary is not known.

Changes in calcitonin gene-related peptide immunoreactivity in the rat dorsal horn following electrical stimulation of the sciatic nerve.

Immunoreactive staining for calcitonin gene-related peptide (CGRP) in the rat superficial dorsal horn (L4-L5 segments) was examined following electrical stimulation of the sciatic nerve. Electrical stimulation for twenty minutes at an intensity to elicit both A and C fiber volleys resulted in a significant decrease in CGRP immunoreactivity in the medial two-thirds of the dorsal horn on the stimulated side as compared to the non-stimulated side of the spinal cord. The change was most pronounced in the medial area of the dorsal horn, with a 54% decrease in immunostaining density compared to the non-stimulated side. In contrast, when the sciatic nerve was stimulated for the same period at an intensity that stimulates only A fibers, there was no significant difference in the density of immunoreactive staining in the dorsal horn between the non-stimulated and stimulated sides of the spinal cord. These results suggest that activity in a specific population of primary afferent fibers causes a dramatic depletion of CGRP in the dorsal horn.

Afferent fibers in the reproductive system and pelvic viscera of female rats: anterograde tracing and immunocytochemical studies.

Innervation of the female reproductive system provides an important signal for a variety of neuroendocrine reflexes and behaviors in the female rat. Although some studies suggest that afferent feedback from the gonads is involved in the hypothalamic control of gonadal function and pituitary hormone release, the extent and function of afferent feedback from the gonads in these neuroendocrine reflexes has yet to be clarified. Deafferentation studies have provided only partial support for the afferent control of the gonads. Some studies even suggest functional asymmetries in the neural control of the gonads, but knowledge regarding the neuroanatomical substrate for these possible neurogonadal interactions remains incomplete. Studies with retrograde tract tracers indicate that the ovaries receive a substantial afferent supply from lower thoracic-upper lumbar dorsal root ganglia. Despite stringent precautions to prevent diffusion of tracers following injections into the ovary or related nerves, many of the retrogradely labeled cell bodies identified by these studies may represent an overestimation of the extent of afferent innervation. We have reexamined the afferent innervation of the female reproductive tract by means of the anterograde transport of horseradish peroxidase (HRP) from thoracic, lumbar and sacral dorsal root ganglion to pelvic visceral organs and have studied the effects of unilateral ganglionectomy on substance P containing fibers in the ovary, oviduct and uterus. The neuroanatomical results show that the T13 and L1 dorsal root ganglia provide major afferent innervation to the cranial portion of the reproductive tract and the L6 and S1 dorsal root ganglia provide primary afferent fibers to the caudal portion of the reproductive tract as well as the bladder, rectum and perineum.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of MK-801 on rat primary afferent neurons and fibers.

Recent evidence suggests that glutamate and its N-methyl-D-aspartate (NMDA) receptor may participate in regulating neurite morphology and peptide expression. A previous study from this laboratory showed that treatment with the NMDA receptor antagonist, MK-801, induced an apparent increase in the density of calcitonin gene-related peptide (CGRP)-immunoreactive primary afferent fibers in the dorsal spinal cord of the rat. The present study was undertaken to extend this work by: 1) quantifying the MK-801-induced increase in CGRP immunostaining in the dorsal grey commissure/medial dorsal horn region and 2) examining the effect of MK-801 on the number of CGRP-immunoreactive primary afferent cell bodies in lumbar dorsal root ganglia. Following 7 days of MK-801 treatment, a significant increase (p less than 0.001) in CGRP immunostaining was observed in the dorsal grey commissure/medial dorsal horn. However, after MK-801 treatment, no significant difference was noted in the numbers of CGRP-immunoreactive primary afferent cell bodies in dorsal root ganglia. These data suggest that MK-801 produces significant alterations in the intraspinal projection of CGRP-immunoreactive fibers without inducing immunocytochemically detectable CGRP within a new population of primary afferent neurons.

A simple approach to ranking a group of aggregated fuzzy utilities.

When ranking a large quantity of fuzzy numbers, the efficiency, accuracy, and effectiveness of the ranking process is critical. The paper considers the application of "alpha-cut" and "fuzzy arithmetic operations" to the fuzzy weighted average (FWA) method which can be used to rank aggregated fuzzy utilities (or generalized fuzzy numbers). The purpose of this application is to make the method easier to program and the data easier to manipulate, which results in a more practical method for fuzzy decisions.

Dietary supplementation of conjugated linoleic acid in horses increases plasma conjugated linoleic acid and decreases plasma arachidonic acid but does not alter body fat.

Studies using dietary supplementation of eicosapentaenoic and docosahexaenoic fatty acids (FA) in horses report inconsistent anti-inflammatory results but consistently report an increase in plasma arachidonic acid (C20:4), the major substrate of cyclooxygenase (COX) II inflammatory pathway. Conjugated linoleic acid has shown anti-inflammatory effects in laboratory and food animal species, but effects of CLA supplementation in horses have not been reported. Our objective was to determine the effects of CLA supplementation on plasma CLA and C20:4 and body fat in healthy horses at maintenance. In a crossover study, 12 mature mares were blocked by breed, age, and BCS and separated into 2 treatment groups (n = 6/group). Groups were fed CLA and corn oil (CO; isocaloric control) for two 6-wk feeding periods, separated by a 4-wk period during which treatment was withheld. Corn oil or CLA supplement (55% mixed CLA isomers) was incorporated into diets at 0.01% BW/d. Mares were fed individually and restricted to dry lots to control forage intake. Rump fat thickness (RFT), BW, and BCS were measured before (d 0) and after (d 42) each feeding period. Blood was collected on d 0, 14, 28, and 42 of each 6-wk period for GLC analysis of plasma CLA isomers (cis-9, trans-11; trans-10, cis-12; and trans-9, trans-11) and C20:4. An ANOVA was conducted to compare the response of RFT, BW, and BCS of CLA-treated and control mares. A mixed methods analysis with repeated measures was used to detect differences in plasma FA concentrations. There were no differences in BW, RFT, or BCS between treatment groups. All CLA isomers present in the CLA supplement were greater in plasma of horses fed CLA compared with controls (P < 0.01). Additionally, plasma concentrations of C20:4 were decreased in horses fed CLA (P < 0.05). This decline in C20:4 may impact the COX II pathway and warrants further investigation. These results suggest that in an equine model, dietary CLA increases circulating concentrations of supplemented CLA isomers and decreases circulating C20:4. Examining physiological effects of CLA supplementation in horses at varying levels of growth, exercise, and progression of joint disease may offer insight to potential benefits of CLA in the horse.

[Orbital tumours in children and adolescents. Differential diagnostics and clinical symptoms]. / Orbitale Raumforderungen im Wachstumsalter. Differenzialdiagnostik und Krankheitsbilder.

BACKGROUND: The aim of this retrospective study was to determine the incidence of and surgical strategies used for treating orbital tumours in children and adolescents. PATIENTS AND METHODS: A retrospective chart review was carried out for 22 patients between the ages of 2 weeks and 18 years who were treated in the years 1997-2007 for suspected space-occupying orbital lesions. The specific investigation methods, therapy, and treatment results were described. RESULTS: The most frequent orbital lesions in the children in this study were mesenchymal tumours; the second most common were dermoids. Nineteen patients were surgically treated. Three children with malignant tumours underwent radiochemotherapy after surgery, and two died in the first year after diagnosis. Two other children with benign tumours suffered recurrence of their disease. CONCLUSION: The variety of orbital diseases in paediatric patients includes benign and malignant tumours as well as nonneoplastic lesions. The treatment of orbital disease is a diagnostic and therapeutic challenge for the attending physician.

Substance P- and vasoactive intestinal polypeptide (VIP)-immunoreactive nerve fibers in relation to ovarian postganglionic perikarya in para- and prevertebral ganglia: evidence from combined retrograde tracing and immunocytochemistry.

Retrograde neuronal tracing with the fluorescent dye True Blue and immunocytochemistry were utilized to examine postganglionic sympathetic neurons in para- and prevertebral ganglia projecting to the rat ovary. Perikarya in both ganglia were labeled with True Blue after application of the tracer to either the superior ovarian or ovarian plexus nerve. After application of True Blue to the superior ovarian nerve, 17% of the labeled cells in paravertebral ganglia were immunoreactive for vasoactive intestinal polypeptide. In contrast, after application of True Blue to the ovarian plexus nerve, approximately 1% of the labeled cells in paravertebral ganglia were immunoreactive for the same polypeptide. Some vasoactive intestinal polypeptide-immunoreactive perikarya in paravertebral ganglia were not labeled with True Blue. In some cases, substance P- and vasoactive intestinal polypeptide-immunoreactive fibers were closely apposed to True Blue-labeled perikarya in para- and prevertebral ganglia. Paravertebral vasoactive intestinal polypeptide-immunoreactive perikarya projecting to the ovary presumably participate directly in the control of various ovarian functions. Substance P- and vasoactive intestinal polypeptide-immunoreactive fibers closely apposed to perikarya projecting to the ovary may participate indirectly in the control of various ovarian functions by affecting the activity of ovarian postganglionic neurons.

Simple thin-layer chromatographic method for the investigation of six related compounds in flurazepam hydrochloride and its capsules.

A simple and rapid thin-layer chromatographic procedure is presented for the separation and tentative identification of flurazepam hydrochloride and its six related impurities in bulk samples and capsules. A methanolic sample solution is applied to a plate of silica gel containing a fluorescent indicator, which is developed once with a basic quaternary solvent system. Spots are visible under a short wavelength UV lamp, with a limit of detectability ranging from about 62.5 ng (related compound B) to 500 ng (related compound C). The proposed procedure shows several advantages over the related compounds test of the United States Pharmacopeia.