Histochemical and morphometric investigations of the musculature of forelimb mass of sheep with reference to its function. 1. Bending and extension of elbow joints]

Muscle tissue was removed from the extensors and flexors of the elbow joint of six male sheep (180 days old) and stained for NADH tetrazolium oxidoreductase and myofibrillar ATPase after preincubation at pH 4.3 in order to identify three fiber types: slow twitch oxidative (STO), fast twitch oxidative (FTO) and fast twitch glycolytic fibers (FTG). The medial head of the M. triceps brachii and the anconaeus muscle had the largest fibers (> 50 and 60 microns). The smallest muscle fibers (35-43 microns) were found in the dorsal part of the long head of the triceps muscle. The medial head of the triceps muscle and the anconaeus muscle possessed a very high percentage of STO-fibers (90 and 100%) and FTG-fibers were absent in these muscles. In the other extensors and flexors of the elbow joint the STO-percentage amounted to less than 30%. The dorsal part of the long head of the triceps muscle contained only 13% STO-fibers, but had the highest percentage of FTG-fibers (49%), which is representative of fast-muscles. The muscles of the elbow joint perform both static and dynamic functions. The medial head of the triceps brachii muscle and the anconaeus muscle possess the complement of enzymes which permits them to fulfil the work of extensors in the standing position. Therefore, they are typical of antigravity muscles. The histochemical structure of the other extensors and flexors reflect their function in motion. The lateral and long head of triceps muscle oppose the flexors and extend the elbow joint of the raised limb in the swing phase, during the landing phase they also function to support the other extensors of the elbow.     

Histochemical studies of human stomach biopsies with special reference to hydrolases

Peptidases, phosphatases, glycosidases, non-specific esterases, succinate dehydrogenase, cholinesterases, and carbohydrate components were studied in bioptic material of the normal and diseased human stomach using well established older, modified older, and new qualitative histochemical methods. For the first time, an enzyme pattern is reported for all regions of the human mucosa. Local and regional enzyme histochemical differences existed between the cardiac, fundic, body, and pyloric mucosa. Differences were absent, however, in the same region, and no differences were found between the anterior and posterior wall and the large and small curvature of the stomach. In cases of histologically less severe gastritis as a rule, enzyme histochemical changes were not found. They were numerous, however, in biopsies of patients with severe gastritis; only amino-peptidases A and M were unchanged. Dipeptidyl peptidase IV was absent; gamma-glutamyl transpeptidase exhibited individual differences. Alkaline phosphatase occurred in the pericapillary stroma and adenosine phosphates were not hydrolysed in atrophic glandular epithelia. Activity increases of lysosomal dipeptidyl peptidase I and beta-D-glucuronidase were typical for inflammatory infiltration processes of the gastric mucosa. Severe atrophy was accompanied by an activity decrease of glandular non-specific esterases, dipeptidyl peptidase II, and beta-N-acetyl-D-glucosaminidase and an activity decrease of the stromal peptidases and glycosidases. Enzyme activity was absent in the gastric glands proper in cases of total atrophy. An increase in macrophage number was primarily linked with an increase in acid phosphatase activity. Alkaline phosphatase, aminopeptidase M and gamma-glutamyl transpeptidase activities were enhanced in malignant neoplasms. High activities of all peptidases and alkaline phosphatase were found in the brush border of surface epithelial cells in cases of intestinal metaplasia. Except for dipeptidyl peptidase I and II, the enzyme pattern corresponds to that of small intestinal enterocytes. Compared with histological routine procedures for gastric diagnosis and assessment of the course enzyme histochemical methods deliver additional information; practically, however, the enzyme histochemical analysis of gastric biopsies are only useful in special cases.     

Morphological and morphometric studies of the airways of sheep with acute airway hypersensitivity to inhaled Ascaris suum.

The airways of 12 sheep with naturally-occurring allergic airway hypersensitivity, six of which had changes in both airway resistance and dynamic lung compliance (Group A) and six of which had changes in only dynamic lung compliance (Group B), were compared quantitatively with six non-reacting sheep (Group C) in order to examine the relation between airway hypersensitivity and various morphological features thought to be related to airway hypersensitivity. Compared to the non-reacting sheep (Group C), the hypersensitive sheep (Groups A and B) had a thinner epithelium in medium bronchi and bronchioles, fewer goblet cells in bronchioles, and greater gland area at most airway levels. The differences of the gland dimensions and the types of mucosubstance between hypersensitive and non-reacting animals were more variable. No significant differences between the three groups were noted with regard to luminal occlusion or epithelial sloughing and squamous metaplasia. Although there was a positive association between epithelial thickness and goblet cell density in the small airways, the development of allergic airway hypersensitivity in sheep may occur in the absence of major morphological changes in the airway epithelium.     

Absence of recurrent axon collaterals in motoneurones to the extrinsic digit extensor muscles of the cat forelimb.

Forelimb alpha-motoneurones were intracellularly recorded in anaesthetized cats and iontophoretically filled with horseradish peroxidase (HRP). All motoneurones to the elbow flexors, elbow extensor and to the extensor carpi radialis muscles displayed in parallel homonymous recurrent inhibitory postsynaptic potentials (RIPSPs) and axon collaterals. Homonymous RIPSPs and axon collaterals were missing in the nuclei to the long digit extensor muscles. Two populations of motoneurones, with and without recurrent axon collaterals, seem to be present in the extensor carpi ulnaris motor nucleus. These results are consistent with the hypothesis that the motoneurones to the extrinsic digit extensors lack a recurrent axonal system. This indicates that the contribution of the recurrent Renshaw systems to motor control may be more complex than hitherto assumed.     

Histochemical studies of intercellular components of salivary gland tumors with special reference to glycosaminoglycan,laminin and vascular elements

Summary In 41 salivary gland tumors, the characteristics of the intercellular components and vascular endothelial cells were surveyed by immunohistochmical staining for laminin and factor VIII-related antigen (VIII R:Ag), and by mucopolysaccharidase-digestion for glycosaminoglycan (GAG). In myxomatous areas of pleomorphic adenomas, small vessels (diameter 6.5 ± 0.11 µm) were frequent and found to be negative or weakly positive by VIIIR:Ag staining although endothelial cells were clearly positive for VIIIR:Ag in capsule surrounding the tumor tissues. Alcian blue stainability was diminished by treatment with both Streptomyces hyaluronidase and chondroitinase. By laminin staining, a vascular pattern was clearly detected, but the majority of tumor cells were not stained. In adenomatous areas, the basement membrane-like linear laminin-staining reaction was observed to be weak and inconsistent around some tumor cell nests. However, in adenoid cystic carcinomas, laminin-positivity was much more intense than in other tumors such as pleomorphic adenoma, mucoepidermoid tumor and adenocarcinoma. In cylindromatous areas, the inner luminal surface in the pseudocysts was markedly positive for laminin, and there was weak positivity around tumor cell nests having a trabecular pattern. By immunoelectron microscopy, a juxtacellular network of replicated basal lamina of tumor cells which lined the inner surface of pseudocysts was positive for laminin. Alcian blue-positivity in the pseudocyst was abolished with heparitinase and chondroitinase, but not with hyaluronidase.     

Neuronal projections to the lateral retrochiasmatic area of sheep with special reference to catecholaminergic afferents: immunohistochemical and retrograde tract-tracing studies

The retrochiasmatic area contains the A15 catecholaminergic group and numerous monoaminergic afferents whose discrete cell origins are unknown in sheep. Using tract-tracing methods with a specific retrograde fluorescent tracer, fluorogold, we examined the cells of origin of afferents to the retrochiasmatic area in sheep. The retrogradely labeled cells were seen by observation of the tracer by direct fluorescence or by immunohistochemistry with specific antibodies raised in rabbits or horses. Among the retrogradely labeled neurons, double immunohistochemistry for tyrosine hydroxylase, dopamine-beta-hydroxylase, and serotonin were used to characterize catecholamine and serotonin FG labeled neurons. The retrochiasmatic area, which included the A15 dopaminergic group and the accessory supraoptic nucleus (SON), received major inputs from the lateral septum (LS), the bed nucleus of the stria terminalis (BNST), the thalamic paraventricular nucleus, hypothalamic paraventricular and supraoptic nuclei, the perimamillary area, the amygdala, the ventral part of the hippocampus and the parabrachial nucleus (PBN). Further, numerous scattered retrogradely labeled neurons were observed in the preoptic area, the ventromedial part of the hypothalamus. the periventricular area, the periaqueductal central gray (CG), the ventrolateral medulla and the dorsal vagal complex. Most of the noradrenergic afferents came from the ventro-lateral medulla (Al group), and only a few from the locus coeruleus complex (A6/A7 groups). A few dopaminergic neurons retrogradely labeled with flurogold were observed in the periventricular area of the hypothalamus. Rare serotoninergic fluorogold labeled neurons belonged to the dorsal raphe nucleus. Most of these afferents came from both sides of the brain, except for hypothalamic supraoptic and paraventricular nuclei. In the light of these anatomical data, we compared our results with data obtained from rats, and we discussed the putative role of these afferents in sheep in the regulation of several specific functions in which the retrochiasmatic area may be involved, such as reproduction.     

Histological studies on the inner surfaces of the articular cavities of human temporomandibular joints with special reference to arthroscopic observations

Histological findings relating to the arthroscopic anatomical features in the inner surfaces of human temporomandibular joints are described. Articular surface covering the bony surface is histologically fibrocartilage. The articular surface of the articular disk is histologically composed mainly of tightly packed collagen fibers, but partly of fibrocartilage which is considered to be developed in the area where presumably maximal stress is given during the joint movement. The synovial membrane consist of 2 layers. The superficial one is the lining cell layer formed by single or multiple rows of cells with from ovoid to cuboidal shapes. The deep one is a capillary rich loose connective tissue layer. The present study suggests that arthroscopic views well reflect histological features of the inner wall of the articular cavity of the temporomandibular joint.     

Comparative anatomy, homologies and evolution of the pectoral and forelimb musculature of tetrapods with special attention to extant limbed amphibians and reptiles

The main aim of the present work is to synthesize the information obtained from our dissections of the pectoral and forelimb muscles of representative members of the major extant taxa of limbed amphibians and reptiles and from our review of the literature, in order to provide an account of the comparative anatomy, homologies and evolution of these muscles in the Tetrapoda. The pectoral and forelimb musculature of all these major taxa conform to a general pattern that seems to have been acquired very early in the evolutionary history of tetrapods. Although some muscles are missing in certain taxa, and a clear departure from this general pattern is obviously present in derived groups such as birds, the same overall configuration is easily distinguishable in these taxa. Among the most notable anatomical differences between the groups, one that seems to have relevant evolutionary and functional implications, concerns the distal insertion points of the forearm musculature. In tetrapods, the muscles of the radial and ulnar complexes of the forearm are pleisomorphically mainly inserted onto the radius/ulna or onto the more proximal carpal bones, but in mammals some of these muscles insert more distally onto bones such as the metacarpals. Interestingly, a similar trend towards a more distal insertion of these muscles is also found in some non‐mammalian tetrapod taxa, such as some anurans (e.g. Phyllomedusa). This may be correlated with the acquisition of more subtle digital movement abilities in these latter taxa.     

From fish to modern humans – comparative anatomy, homologies and evolution of the pectoral and forelimb musculature

In a recent study Diogo & Abdala [(2007 ) J Morphol  268 , 504–517] reported the results of the first part of a research project on the comparative anatomy, homologies and evolution of the pectoral muscles of osteichthyans (bony fish and tetrapods). That report mainly focused on actinopterygian fish but also compared these fish with certain non-mammalian sarcopterygians. This study, which reports the second part of the research project, focuses mainly on sarcopterygians and particularly on how the pectoral and forelimb muscles have evolved during the transitions from sarcopterygian fish and non-mammalian tetrapods to monotreme and therian mammals and humans. The data obtained by our own dissections of all the pectoral and forelimb muscles of representative members of groups as diverse as sarcopterygian fish, amphibians, reptiles, monotremes and therian mammals such as rodents, tree-shrews, colugos and primates, including humans, are compared with the information available in the literature. Our observations and comparisons clearly stress that, with regard to the number of pectoral and forelimb muscles, the most striking transition within sarcopterygian evolutionary history was that leading to the origin of tetrapods. Whereas extant sarcopterygian fish have an abductor and adductor of the fin and a largely undifferentiated hypaxial and epaxial musculature, extant salamanders such as Ambystoma have more than 40 pectoral and forelimb muscles. There is no clear increase in the number of pectoral and forelimb muscles within the evolutionary transition that led to the origin of mammals and surely not to that leading to the origin of primates and humans.     

Immunopathologic studies of renal glomerular change in liver cirrhosis with special reference to its pathogenesis.

The kidneys from 62 proven liver cirrhosis cases were examined by immunofluorescence (IF), and 94% of the cases were positive for some immune reactants deposition. Combined deposition of immunoglobulin(s), both or each one of C1q and C4, and further with C3 were observed in about 70% of all IF positive cases. The morphological alterations of the glomeruli correlated with the intensities of regional immune reactants depositions. Guinea pig C3 was frequently activated in vitro on the glomeruli of these cases. Immune reactants depositions in the glomeruli appeared to form immune complex locally. Smooth muscle and liver cell antigens in the immune complex at the glomeruli were examined by indirect method of IF using monospecific antibodies and positive cases concerning each antigen were found in about 1/3 of the kidneys from 21 liver cirrhosis cases. These facts suggest that the high rate of the occurrence of immune complex deposition type glomerulo-nephritis may be due to the glomerular deposition of some autoantigen-antibody complexes including smooth muscle and liver cell antigens.     

Development of the spinal nerves in the mouse with special reference to innervation of the axial musculature

Development of the mouse spinal nerves was studied. On E11 (11th day of gestation), the primitive spinal nerve fascicle extended ventrally in the anterior half of the sclerotome. Spinal nerves in the forelimb region united with each other to form the primitive brachial plexus. Their terminal segment was covered by a peculiar cell mass. On E12, five primary branches developed along the primitive spinal nerve trunk. The ramus dorsalis was originally a cutaneous nerve, supplying two series of branches to the skin of the back. The medial series was derived from the dorsal ramus of C2–C8, and the lateral series from C8 and the more caudal dorsal rami. Nerves of the former series took the presegmental course through the intermyotomic space, while those of the latter the postsegmental course. The ramus cutaneus lateralis was a nerve that took the presegmental course to become cutaneous. The ramus intercostalis externus was a muscle branch whose distribution was restricted within the segment. The ramus anterior was a muscle branch from the end of the primitive spinal nerve trunk. The ramus visceralis connected a thoracic nerve with the para-aortic sympathetic cell cord. On E13–16 the ramus anterior secondarily gave off a cutaneous branch (ramus cutaneus anterior). The ramus intercostalis externus extended ventrally deep to the intercostalis externus muscle, crossing just caudal to the ramus cutaneus lateralis that secondarily gave off branches to the obliquus externus abdominis muscle.     

Vasculitis and its renal manifestation with special reference to anti-neutrophil cytoplasmic antibody]

The nomenclature and classification of vasculitis have been problematic, with each researcher proposing their own system. Since the discovery of anti-neutrophil cytoplasmic antibody and the consensus conference on the nomenclature of vasculitis in Chapel Hill, order has been brought to the chaos to some extent. Measurement of P-ANCA and C-ANCA in patients' sera has exerted an immeasurable influence on the clinical aspects of vasculitis, since they are shown to be valuable for the diagnosis and assessment of the vasculitic activity in most cases. Pathogenetic significance is also given to ANCA. ANCA has been reported to release myeloperoxidase(MPO) and proteinase-3 antigens from neutrophils activated by inflammatory cytokines. When both MPO antigen in serum as well as ANCA were measured serially in the same patients, however, clinical condition was better reflected by the amount of serum MPO than the titer of ANCA in some patients. Thus, the amount of serum MPO increased without concomitant rise in ANCA titer in exacerbation of vasculitis in one patient. In two more, high serum titers of ANCA and low MPO antigen in the active phase of vasculitis were reversed when pulmonary hemorrhage occurred, and fatal outcome ensued. It could be speculated from these observations that some ANCA might be directed against the active sites of MPO and even suppress the vasculitic activity through blocking MPO. Since definitive evidence has not been obtained so far on the pathogenetic role of ANCA, the clinical significance of ANCA should be addressed at the same level as anti-dsDNA antibody in systemic lupus erythematosus: aid for diagnosis and disease activity.