Diverse changes in fibre type composition of the human lateral pterygoid and digastric muscles during aging

The fibre type composition of the superior and inferior portions of the human lateral pterygoid and the anterior and posterior bellies of the digastric muscles of five elderly subjects (mean age 73 years) was studied by morphological and enzyme-histochemical methods. Both muscles showed significant age-related changes in fibre type composition as compared with previous data for young adults. In the lateral pterygoid we observed a large proportion of type IIA fibres, which are rare or absent in young adults, and muscle fibre atrophy and an increased variability in fibre diameter. The digastric muscle of elderly showed a decrease in the proportion of type IIB fibres. The only difference in age-related changes between muscle portions was found in the lateral pterygoid with fibre atrophy in its inferior portion. Both the lateral pterygoid and digastric muscles are known to be active in mandibular depression (jaw opening) and horizontal positioning of the mandible. The present results and previous data from young adults show that the lateral pterygoid and digastric muscles differ not only in fibre type composition, but also in muscular changes following aging. This suggests that, even if they are simultaneously active, they fulfill different, specific tasks in natural jaw function. The differences in age-related changes in fibre type composition between these two muscles indicate that mechanisms underlying their alterations during aging are muscle specific. The results indicate that, although nerve supply and developmental history are essential for fibre composition of skeletal muscles, functional tasks and demands are of major importance.     

Isolation of sex-specific cDNAs from fetal mouse brain using mRNA differential display and representational difference analysis

Comparing female and male brain structures reveals a variety of sex differences in many vertebrates. These differences are manifested throughout the brain, in regions such as the hypothalamus, the preoptic area and the amygdala. Some are thought to be induced during the fetal period by the effect of steroid hormones produced in the gonads. It is well-established that fetal androgens, probably through the conversion to estrogen by the enzyme aromatase, masculinize the nervous system and set adult mounting behavior in rodents. However, less is known about molecular mechanisms involved in gender-specific development of the brain. We have taken a broad approach to isolate sex-specific genes from fetal brain. mRNAs from 18.5 days post-coitum (dpc) female and male mouse brain were screened with the classical and the recently developed signal peptide differential display (SPDD) and with representational difference analysis of cDNA (cDNA-RDA). Two sex-specific cDNAs were isolated, F29 and M17, corresponding to the female-specific Xist gene and the male-specific Smcy gene, respectively.     

Delta-opioid receptor immunoreactivity on astrocytes is upregulated during mitosis

Endogenous opioid peptides and opioid receptors are expressed by brain cells early during normal development, and exogenous opiate exposure in this period is known to affect brain cell proliferation and maturation. Despite the abundant evidence that opioids affect brain development, little is known about the mechanisms involved. In this study cortical astrocytes in primary culture were examined immunohistochemically by using antibodies against the opioid receptors. The immunoreactivity for delta-opioid receptors was strongly upregulated during mitosis with an increase in immunostaining that started in early prophase and lasted through the M-phase to cytokinesis. Similar effects could not be observed when antibodies against the mu- or kappa-opioid receptor subtypes were used. Cultured neurons and microglia presented a strong and homogenous immunostaining for the delta-opioid receptor and no further upregulation of immunoreactivity could be detected in these cells. The presence of functional delta-opioid receptors on the mitotic astrocytes was verified by using microspectrofluorometry for detection of delta-opioid agonist induced changes in intracellular free calcium concentrations ([Ca2+]i). In these experiments fluo-3/AM incubated cells showed a rapidly induced delta-opioid agonist (DPDPE, 10(-6) M) evoked increase in [Ca2+]i. These results suggest an upregulation of the delta-opioid receptors that could represent a mechanism involved in the response to opioids in the developing brain.     

Analysis of CSF amino acids in young patients with generalised refractory epilepsy during an add-on study with lamotrigine

The effect of add-on administration of lamotrigine (1-12 mg/kg per day, 2-12 months) on the levels of neurotransmission related amino acids including gamma-aminobutyric acid (GABA), glutamate, aspartate, glycine and antiepileptic drugs (AEDs) in lumbar cerebrospinal fluid (CSF) was studied in 22 children and young adults with generalised therapy resistant epilepsy. Two lumbar punctures were performed, one prior to, and one following a mean of 5 months (2-12 months) of lamotrigine treatment. Lamotrigine decreased seizure incidence and severity in 12 of the 22 patients without influencing CSF GABA, glutamate, aspartate or glycine levels. Lamotrigine did not alter the concentrations of AEDs in CSF or plasma. However, CSF GABA levels were 86% higher in those patients also treated with gamma-vinyl-GABA (vigabatrin, GVG) compared with patients treated with other combinations and this was not altered by co-medication with lamotrigine. The proposed mechanism of action of lamotrigine, namely that it may inhibit glutamate release in the CNS, is not reflected by changes in CSF glutamate levels. The present findings indicate that CSF GABA, glutamate, aspartate and glycine levels may not be useful as in vivo neurochemical markers in young patients responding to the therapeutic dose of lamotrigine used in this study.     

Developmental neurotoxicity of four ortho-substituted polychlorinated biphenyls in the neonatal mouse

The objective of the present study was to investigate whether neonatal exposure to single PCB (polychlorinated biphenyl) congeners 2,4,4'-trichlorobiphenyl (IUPAC 28), 2,2',5,5'-tetrachlorobiphenyl (IUPAC 52), 2,3',4,4',5-pentachlorobiphenyl (IUPAC 118) and 2,3,3',4,4',5-hexachlorobiphenyl (IUPAC 156) when given as one single dose (0.7-14 μmol/kg body weight per os) to 10-day-old male NMRI mice could induce persistent neurotoxic effects in the adult animal. Furthermore, to ascertain whether behavioural aberrations, both in spontaneous behaviour and in learning and memory function, were followed by changes in the cholinergic and/or the dopaminergic system. It was found that neonatal exposure to lightly chlorinated ortho-substituted PCBs, 2,4,4'-tri- and 2,2',5,5'-tetrachlorobiphenyls, can induce persistent aberrations in spontaneous behaviour. Neonatal exposure to 2,2',5,5'-tetrachlorobiphenyl also affected learning and memory functions in the adult animal. In the animals showing a deficit in memory and learning function, the cholinergic nicotinic receptors in the cerebral cortex were affected. Exposure to 2,3',4,4',5-penta- and 2,3,3',4,4',5-hexachlorobiphenyl, mono-ortho congeners ('co-planar-like'), in the same dose range did not cause any significant change in the investigated behavioural variables, spontaneous and swim-maze behaviour.     

Effects of α2-adrenoceptor agonists on locus coeruleus firing rate and brain noradrenaline turnover in N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ)-treated rats

Summary Previous studies have shown that a low dose of the alkylating compound N-ethoxycarbonyl-2-ethoxy1,2-dihydroquinoline (EEDQ) reduces the density of ₂-adrenoceptors in rat cerebral cortex and antagonizes the effects of an ₂-adrenoceptor agonist on noradrenaline release in rat cortical slices. In the present study, a corresponding dose of EEDQ (1 mg/kg, s. c., 24 h) was shown to reduce the effect of the ₂-adrenoceptor agonists clonidine and guanfacine on noradrenaline turnover in rat brain while not affecting the inhibitory effect of clonidine on locus coeruleus (LC) cell firing. When considerably higher doses of EEDQ were administered (10 and 20 mg/kg, s. c., 24 h) not only the biochemical but also the electrophysiological effects of clonidine were markedly reduced (or even reversed). The data support the notion that EEDQ decreases the responsiveness of brain ₂-adrenergic receptors; moreover, they indicate that ₂-adrenoceptors regulating LC activity are characterized by a larger receptor reserve or are less sensitive to the influence of alkylation than are the population of ₂-adrenoceptors regulating noradrenaline utilization. Send offprint requests to G. Engberg, at the above address     

Pancreatic tumors in multiple endocrine neoplasia type 1: Clinical presentation and surgical treatment

Among 33 patients with endocrine pancreatic tumors due to multiple endocrine neoplasia type 1 (MEN-1), 19 (58%) patients had hypergastrinemia, 7 (21%) patients had hyperinsulinism, and 7 (21%) patients had clinically non-functioning lesions. At least one gross tumor was found in all patients undergoing pancreatic surgery, including those with negative localization studies prior to operation. The patients also had additional macroscopic tumors as well as numerous microadenomas, and the lesions frequently were positive for immunostaining with multiple hormones, mainly pancreatic polypeptide, insulin, glucagon, and somatostatin. Duodenal endocrine lesions were found in 4 of 5 investigated patients and stained with gastrin and somatostatin antibodies. Distal, mainly subtotal pancreatic resection, was performed in 18 patients, eventually combined with caput tumor enucleation or duodenotomy, while a few patients underwent only tumor enucleation or a Whipple procedure. The long-erm outcome of operation was most favorable in patients with hyperinsulinism; only 1 patient had clinical recurrence. Patients with hypergastrinemia experienced only transitory lowering of serum gastrin values after pancreatic surgery and 47% of them had or developed metastases. Such tumor spread was seen in 57% of the patients with non-functioning lesions. Nine patients died from progressive tumor disease during follow-up. Consistent with previous studies, we found that surgery is indicated in MEN-1 patients with hyperinsulinism even if a lesion is not visualized by radiology. In addition, these indications should be extended to also include patients with only biochemical markers of disease, including elevations of gastrin, as these indicate the presence of gross tumors. This strategy should be applied especially in patients with aggressive family histories to possibly reduce the risk of malignant tumor progression.

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Resumen Entre 33 individuos con tumores pancréaticos endocrinos como componente del síndrome de neoplasia endocrina múltiple tipo 1 (NEM-1), 19 pacientes (58%) tenían hipergastrinemia, 7 (21%) hiperinsulinismo y 7 (21%) lesiones clínicas no funcionantes. En la totalidad de los pacientes sometidos a cirugía pancreática fue hallado por lo menos un tumor, incluso en aquellos con examenes de localización negativos anteriores a la operación. Estos pacientes también albergaban tumores macroscópicos, así como numerosos microadenomas; con frecuencia las lesiones demostraron inmunocoloración con diferentes hormonas, principalmente polipéptido, insulina, glucagón y somatostatina. Se encontraron lesiones endocrinas duodenales en 4 de cada 5 pacientes investigados, las cuales colorearon con gastrina y anticuerpos a la somatostatina. Se practicó resección pancreática distal (principalmente resección subtotal) en 18 pacientes, eventualmente combinada con enucleación del tumor (cuando éste se hallaba ubicado en la cabeza del páncreas) o duodenectomía; solamente unos pocos pacientes fueron sometidos a simple enucleación del tumor o al procedimiento de Whipple. El resultado a largo plazo fue más favorable en los pacientes con hiperinsulinismo, puesto que sólo uno presentó recurrencia clínica. Los pacientes con hipergastrinemia exhibieron apenas una disminución transitoria de los valores de gastrina sérica luego de la cirugía pancreática. Cuarenta y siete por ciento del conjunto tuvo o desarrolló metástasis, en tanto que la extensión local del tumor se presentó en 57% de los casos con lesiones no funcionantes. Nueve pacientes murieron por progresión de la neoplasia en el curso del seguimiento. En acuerdo con sugerencias previas, se considero quo la cirugía está indicada en pacientes con NEM-1 e hiperinsulinismo, aún en los casos en que no se visualiza radiológicamente la lesión, pero que la indicación puede ser ampliada para incluir también pacientes con sólo marcadores bioquímicos, tales como niveles elevados de gastrina, indicativos de la presencia de tumores macroscópicos. Esta estrategia debe ser aplicada principalmente en aquellos pacientes con historia familiar agresiva, con lo cual tal vez se reduce el riesgo de progesión maligna del tumor.

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Résumé Parmi 33 patients ayant une tumeur pancréatique endocrine due à une néoplasie endocrine multiple de type 1 (MEN-1), 19 (58%) avaient une hypergastrinémie, 7 (21%) un hyperinsulinisme et 7 (21%) une lésion cliniquement muette. On a mis en évidence au minimum une grosse tumeur chez tous les patients, y compris chez ceux dont les examens préopératoires de dépistage tumoral étaient négatifs. Les patients étaient également porteurs de tumeurs macroscopiques et de nombreux microadénomes. Les lésions montraient souvent un immunomarquage positif pour de multiples hormones, principalement le polypeptide pancréatique, l'insuline, le glucagon et la somatostatine. Des lésions endocrines duodénales furent retrouvées chez 4 des 5 patients explorés; elles montraient un immunomarquage avec les anticorps angigastrine et anti-somatostatine. Une résection pancréatique distale, le plus souvent subtotale, a été réalisée chez 18 patients. Elle était éventuellement complétée par une énucléation tmorale de la tête ou par une duodénotomie. Peu de patients ont bénéficié d'une simple énucléation ou d'une intervention de Whipple. L'évolution postopératoire à long terme a été plus favorable en cas d'insulinome puisque seul un patient a eu une récidive clinique. Les patients atteints de gastrinome n'ont présenté que transitoirement une diminution des taux sériques de gastrine après la chirurgie pancréatique. Quarante sept pour cent de ces patients avaient ou ont développé des métastases contre 57% des patients porteurs de lésions sans traduction clinique. Neuf patients sont décédés en raison de l'extension tumorale au cours du suivi. Conformément à des suggestions antérieures, la chirurgie semble indiquée chez les patients atteints de MEN-1 avec hyperinsulinisme même si la radiologie ne visualise pas de lésion. Mais cette indication peut être élargie aux patients dont seuls les paramètres biologiques sont en faveur d'une grosse tumeur (dont l'hypergastrinémie). Cette stratégie pourrait convenir particulièrement aux patients ayant des antécédents familiaux importants; elle permettrait peut-être de réduire le risque d'extension tumorale.

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Presented at the International Association of Endocrine Surgeons in Stockholm, Sweden, August, 1991.     

Vascularization of skin following implantation of an arteriovenous pedicle: implications in flap prefabrication.

In a rat model, a skin flap was fabricated by implantation of a distally ligated arteriovenous pedicle. The femoral artery and vein were implanted as a unit beneath the abdominal skin, a portion of which was later raised as an island flap, based on these vessels. Percentage area of survival, tissue blood flow, and pattern of vascularity were compared in two groups of flaps. In group I, the pedicle to be implanted was dissected with a cuff of surrounding muscle still attached; in group II, the pedicle was skeletonized to the level of adventitia. Flap survival in the two groups was similar (73% vs. 77%), as was skin blood flow (5.4 vs. 5.7 ml/100 g/min). Angiography demonstrated two principal patterns of vascularization: connection between donor and recipient vessels (inoculation), and sprouting and arborization of new vessels. Strengths and limitations of this and other models of flap "prefabrication" are discussed.     

The presence of glutathione in primary neuronal and astroglial cultures from rat cerebral cortex and brain stem

Summary The concentration of the tripeptide glutathione (GSH) was measured in primary cultures of neurons and astroglial cells from rat cerebral cortex and brain stem. The concentration of GSH was found to be approximately 20 nmol/ mg protein in the neuronal culture from the cerebral cortex and ca. 40 nmol/ mg protein in the neuronal brain stem cultures. A GSH concentration of approximately 20 nmol/mg was observed in the astrocyte cultures from both brain regions. The possibility to increase the GSH concentration was tested by incubating the cultures in the presence of the GSH precursor -glutamylcysteine (-GC). In the cultured astrocytes -GC produced a dose-dependent increase in GSH. A similar increase was observed in the neuronal cultures, but this effect failed to reach statistical significance.