Recent progress in human genetics and neurobiology has led to the identification of various mutations in particular myelin genes as the cause for many of the known inherited demyelinating peripheral neuropathies. Mutations in 3 distinct myelin genes, PMP22, P0, and connexin 32 cause the 3 major demyelinating subtypes of Charcot-Marie-Tooth (CMT) disease, CMT1A, CMT1B and CMTX, respectively. In addition, a reduction in the gene dosage of PMP22 causes hereditary neuropathy with liability to pressure palsies (HNPP), while particular point mutations in PMP22 and P0 cause the severe Dejerine-Sottas (DS) neuropathy. A series of spontaneous and genetically engineered rodent mutants for genes for the above-mentioned myelin constituents are now available and their suitability to serve as models for these still untreatable diseases is an issue of particular interest. The spontaneous mutants Trembler-J and Trembler, with point mutations in PMP22, reflect some of the pathological alterations seen in CMT1A and DS patients, respectively. Furthermore, engineered mutants that either over or underexpress particular myelin genes are suitable models for patients who are similarly compromised in the gene dosage of the corresponding genes. In addition, engineered mutants heterozygously or homozygously deficient in the myelin component P0 show the pathology of distinct CMT1B and DS patients, respectively, while Cx32 deficient mice develop pathological abnormalities similar to those of CMTX patients. Mutants that mimic human peripheral neuropathies might allow the development of strategies to alleviate the symptoms of the diseases, and help to define environmental risk factors for aggravation of the disease. In addition, such mutants might be instrumental in the development of strategies to cure the diseases by gene therapy.

There is evidence for lymphatic drainage of interstitial fluid from the brain along perivascular spaces in a number of mammalian species. Ultrastructural studies suggest that there are similar drainage pathways in the human cerebral cortex. Perivascular spaces in the basal ganglia, however, differ from those in the cortex in that they dilate to form lacunes and rarely accumulate beta-amyloid (amyloid angiopathy) in Alzheimer's disease; in the cortex, lacunes are rare but amyloid angiopathy is common. The aim of the present study is to compare the structure of perivascular spaces in the basal ganglia and at the anterior perforated substance with perivascular spaces in the cerebral cortex. Eight postmortem brains from patients aged 23–80 years (mean 68 y) were examined by light microscopy, by scanning and transmission electron microscopy and by direct visualisation of etched paraffin blocks. The results show that arteries in the basal ganglia are surrounded by 2 distinct coats of leptomeninges separated by a perivascular space which is continuous with the perivascular space around arteries in the subarachnoid space. The inner layer of leptomeninges closely invests the adventitia of the vessel wall and the outer layer is continuous with the pia mater on the surface of the brain at the anterior perforated substance. Veins in the basal ganglia have no outer layer of leptomeninges and thus the perivascular space is continuous with the subpial space. The anatomy of the periarterial spaces in the basal ganglia differs significantly from that in the cerebral cortex where there is only a single periarterial layer of leptomeninges. Differences in structure of perivascular spaces around arteries may reflect relative efficiencies in the drainage of interstitial fluid from different sites in the brain. Futhermore, the structure of the perivascular spaces may contribute to the relatively high frequency of lacunes in the basal ganglia, and the low frequency of amyloid angiopathy at this site in Alzheimer's disease.

The encephalomyocarditis virus of the diabetogenic M-strain (EMC-M) is known to cause diabetes in mice. The EMC-M virus has also been shown to cause paresis in some of the infected animals. The clinical features include an acute ascending predominantly motor paralysis, developing within days. This resembles acute idiopathic polyneuritis. The alpha motor neurons would be a possible target for the virus, so two parameters, the total number and the size distribution of motor neurons, were therefore selected for further investigation in 6 mice with neurological involvement and compared with 6 control mice. The optical fractionator method was applied for estimating the total number of motor neurons and the 3D size distribution was estimated using the rotator method in a vertical design. No difference was found in the total number of motor neurons and the size distributions were similar in the 2 groups. This design can be used as a model for the estimation of the total number of motor neurons and their size distribution in other experimental animal models.

Cultures of rat thymic epithelium were used to measure the effect of thymulin secretagogues on dye-coupling and proliferation. Dye-coupling was assessed after the injection of lucifer yellow dextran which cannot permeate the connexin pore of gap junctions and the smaller, permeant cascade blue. In addition to gap junctional communication, larger intercellular bridges were demonstrated by the transfer of lucifer yellow dextran between cells. The extent of intercellular communication was found to be influenced by both cell density and the number of passages. In control cultures, intercellular communication was reduced in cell groups of low (<20 cells/group) or high cell densities (>100 cells/group) compared with groups of 20–60 cells. The highest coupling indices were found in subcultures 20–30. Taking these factors into account, significant decreases in coupling index were observed after pretreatment of test cultures with factors known to influence the secretion of thymulin (5 U/ml interleukin 1 (α and β), 1 μM progesterone, 1 μM oestrogen, 1 μM testosterone, 1 ng/ml adrenocorticotropic hormone, 100 nm rat growth hormone) but 7.5 ng/ml thymulin had no effect on dye-coupling. The nonspecific gap junction uncoupler, octanol, abolished dye-coupling. Cellular proliferation, as measured by the uptake of tritiated thymidine, showed that the same factors that reduced coupling also increased proliferation. None of these factors affected the number of multinucleate cells present, except interleukin-1β which caused a significant reduction in the average number of nuclei per cell. Thus rat thymic epithelium in vitro provides a model for the study of the direct action of factors on cells of the thymic microenvironment.

Methods are described for the semiquantitative analysis of the connective tissue components of human peripheral nerve using light microscopy. General histological preservation was assessed using haematoxylin and eosin staining and the distribution of collagen type IV was investigated using immunohistochemistry. Several techniques were investigated to establish the one giving optimum structural preservation, immunobinding and greatest contrast for image analysis. Frozen sections were unsuitable for this tissue and paraffin wax sections were therefore used. Alcohol fixation was rejected due to poor preservation of the endoneurium, although immunobinding was excellent. Ice-cold formalin fixation for 24 h was found to be adequate for structural preservation and antibody binding, provided that a protease step was introduced. Trypsin was found to be superior to pepsin for exposing collagen type IV epitopes. Of the detection systems investigated indirect immunofluorescence was not suitable due to considerable autofluorescence of the nerve. The avidin-biotin method provided the greatest contrast, and was therefore the detection method of choice for image analysis. The optimum techniques for image analysis were then used on control human sural nerve to ascertain the best comparative method for collagen type IV in the perineurium. A method of semiquantitative analysis is described which takes into account the fact that there is a close linear relationship between collagen content per unit of perineurium and perineurial perimeter as fascicle size increases in peripheral nerve. This means that data from 2 different sample groups can easily be compared, provided that a range of fascicle sizes is analysed in each case.

The distribution of glycine-immunoreactive (glycine-IR) neurons and their associated axon terminals in the rat cuneate nucleus was studied using antiglycine postembedding immunoperoxidase labelling and immunogold staining, respectively. The immunoperoxidase-labelled glycine-IR neurons were widely distributed in the entire rostrocaudal extent of the nucleus. They made up 30.8% (9671/31368) of the neurons surveyed. Quantitative evaluation showed that the percentage of glycine-IR neurons in the caudal level was significantly higher than that in the middle and rostral levels. The glycine-IR neurons were small cells (mean area=198±1.9 μm2, n=2862) with ovoid or spindle-shaped somata. Statistical analysis showed that the size of the glycine-IR neurons in the rostral level was significantly smaller than that in the middle and caudal levels. Immunogold labelled glycine-IR terminals which contained predominantly pleomorphic synaptic vesicles were mostly small (mean area=1.24±0.03 μm2, n=286) and they constituted 24.7% (286/1158) of the total terminals surveyed. They formed axodendritic, axosomatic and axoaxonic synapses with unlabelled elements. It is suggested from this study that glycine is one of the major neurotransmitters involved in the depression of synaptic transmission in the cuneate nucleus.

This study describes the gross anatomy of the alimentary tract of Houbara Bustards (Chlamydotis undulata macqueenii), Kori Bustards (Ardeotis kori), Rufous-crested Bustards (Eupodotis ruficrista) and White-bellied Bustards (Eupodotis senegalensis) maintained in captivity by the National Avian Research Center in the United Arab Emirates. The morphology of the alimentary tract and the proportions of each region were similar in all 4 species. The length of the oesophagus, combined proventriculus and ventriculus, small intestine, and large intestine formed 24.2–28.4%, 7.3–9.7%, 40.5–55.1% and 9.1–14.7% of the total alimentary tract length respectively. Neither crop nor oesophageal enlargement was observed in the birds examined in this study, although male Kori Bustards possessed a saccus oralis in the oropharyngeal cavity. Oesophagi, proventriculi, ventriculi, caeca and large intestine were well developed in all species. The small intestine was shorter than that of other avian herbivores and granivores when compared on a bodyweight basis. The well differentiated stomachs and well developed caeca of the bustards examined in this study are characteristic of omnivores. Analysis of the mean lengths of the alimentary tract components and weight of the liver and pancreas showed sexual dimorphism in cases where male and female data were available for direct comparison.

The disector is the only objective method for quantifying particles of variable size in a given volume. With this method, cell organelles are identified on adjacent sections, but only those present in one section are counted. When counting extremely rare structures in transmission electron microscope sections (physical disector), the usual procedure of counting on electron micrographs is limited for economic reasons (e.g. micrographs highly outnumbering the investigated structures). Hence, to apply this unbiased stereological method, a modification of the physical disector concerning 3 aspects has been developed. (1) The prerequisite of screening large corresponding tissue areas (here ∼65000 μm2) was fulfilled by examining tissue areas along the edges of ultrathin sections. (2) The size of the counting frame was determined by measuring the lengths of the section margins (minus a guard area) by means of a Morphomat. This value was multiplied by the width of the investigated tissue zone, corresponding to the diameter of the electron microscope viewing screen. (3) Disector counting was carried out simultaneously on both sections (bidirectional disector) to improve efficiency. In the present study tiny synaptic bodies (SBs) were quantitated by disector in a rat pineal gland, yielding ∼30 SBs/1000 μm3. By contrast, single section profile counts of SBs amounted to 90 SBs/20000 μm2. Since the presently described adaptation of the disector is time-consuming, it is proposed to determine a proportion factor allowing to estimate number of structures per volume based on single section profile counts. This would decrease the evaluation time by more than 50%.

Complete cross-sections from the femoral midshaft of 180 individuals of known height and weight, aged 21–97 y, from a modern Australian population were examined using automatic video image analysis to quantify total subperiosteal porosity (TSPP). More specifically, the aim was to investigate whether age changes were similar in both sexes in (1) total subperiosteal area (TSPA), cortical area (CA) and medullary area (MA), (2) intracortical porosity (ICP), and (3) the respective contributions to TSPP made by MA and intracortical void area (ICVA). Our findings indicated that both sexes showed a significantly greater height normalised TSPA in the 70s as compared with the 20s. Males had consistently larger bones with a greater height normalised CA. In both sexes CA showed a tendency to increase till the 7th decade and then to decline, more so in females. MA approximately trebled in females and doubled in males over the age range studied. Although ICP also increased, from 4–6% in young adults to over 9% in the elderly, it showed a significant difference between the sexes only in the 3rd decade, being greater in males at this stage. By contrast, TSPP became significantly greater in females (from that recorded in the 3rd decade) by the time they reached the 50s, while in males this did not occur till the 80s. It increased from ∼25% in young adults of both sexes to ∼50% in females and ∼37% in males in their 80s. However, in the elderly there was great variability in both sexes in the appearance of bones from individuals of similar chronological age. Some bones differed little from those in younger subjects, others showed greatly increased ICP, still others displayed reduced cortical widths with low ICP. The femoral midshaft resembles other skeletal sites in that age changes in TSPP are more marked in females than males.

Macrophages are important in inflammatory processes in heart disease and in transplantation rejection. A resurgence of interest in the macrophage has emanated from recent evidence implicating it as an effector cell in atherosclerosis and transplantation rejection. The detailed distribution of the macrophage within the normal human heart is unknown. We quantified macrophage numbers in the different chambers of the heart. Large tissue blocks (1.5–2.0 cm3) were removed from specific sites in 5 ‘normal’ control hearts (2 males, 3 females, age range 19–46 y). Paraffin-embedded sections were stained with a CD68 pan macrophage marker. Positive cells were enumerated within 20 random fields. Results were analysed using a generalised linear modelling method using the Poisson distribution. Macrophages were identified within septa, and often close to blood vessels, in the myocardium, and in the majority of areas in all hearts. Macrophage numbers varied significantly between areas (range 0–6 cells/high power field; P<0.001), and between the 5 hearts analysed (P<0.001). In general, there were significantly more macrophages in the ventricles (RV P<0.01, LV P<0.05), but these differences were affected by heart differences. This study provides a baseline for the range of macrophage numbers within normal hearts, thus enabling comparisons with macrophage numbers within diseased and transplanted hearts.

Ovaries from adult cycling rats were studied from 1600 h on the day of prooestrus to 0700 h on the day of oestrus in order to relate the cyclic hormonal changes to the proliferative activity of preovulatory and postovulatory (i.e. newly-formed corpora lutea) follicles. Proliferative activity was studied by the immunohistochemical demonstration of DNA-incorporated 5-bromodeoxyuridine (BrdU). The proliferative activity of granulosa cells (GC) in large preovulatory follicles showed a centripetal pattern and decreased during prooestrus, reaching a minimum at 2100 h. However, a proliferative wave was found in the GC of preovulatory follicles at 0200 h on the day of oestrus and in those of newly-formed corpora lutea at 0700 h on the day of oestrus. These results suggest that the granulosa cells of preovulatory follicles show maturational changes that followed a different pattern, depending on their location within the follicle, and that the proliferative wave found from 0200 to 0700 h on oestrus is important for the establishment of the number of steroidogenic cells in the cyclic corpus luteum.

Peripheral sensory and autonomic neurons are known to possess insulin receptors. These have been considered to be of the peripheral type, i.e. similar to those of hepatic and fat cells rather than of the brain type which show dual specificity for both insulin and insulin-like growth factor (IGF-1). We have examined the localisation of insulin and IGF-1 receptors in cultured sensory and sympathetic ganglion cells using confocal microscopy and indirect labelling with FITC (fluorescein isothiocyanate) and TRITC (tetramethyl rhodamine isothiocyanate) respectively. We have shown that in cultured U266B1 multiple myeloma cells these receptors display separate localisation, whereas they are colocalised in IM-9 lymphocytes which are known to possess hybrid receptors. We have confirmed the sequestration of insulin and IGF-1 receptors in the cytoplasm of sensory and sympathetic neurons, consistent with a brain-type receptor. The colocalisation of insulin and IGF-1 receptors in sensory and sympathetic ganglion cells is consistent with the view that they are hybrid receptors, similar to those present in the CNS. The function of these receptors, as suggested for the CNS, may be related to trophic support for neurons.

The peptidergic and noradrenergic innervation of rat and human thymus was investigated by immunohistochemistry at the light and electron microscopical level (avidin-biotin-complex, sucrose-phosphate-glyoxylic-acid, and immunogold techniques). The distribution of noradrenergic neural profiles, and positive immunoreactivity for calcitonin gene-related peptide (CGRP), tyrosine hydroxylase (TH) and neuropeptide Y (NPY) is described in female rats during ageing, and in human children. In the neonatal rat thymus, the arteries and septa are well supplied by fine varicose nerves. In older animals (2 wk–1 y) the number of septa and blood vessels increase and consequently also the innervation. No nerves were found in the cortex. Apart from the innervation of the septal areas, immunoreactivity for CGRP and TH was present in thymic cells. Except for the young rats (neonatal–14 d), all rats showed CGRP positivity in subcapsular/perivascular epithelial cells (type 1 cells). All rat thymuses also contained a few TH positive cells in the medulla, which could only be confirmed as epithelial cells (type 6 cells) in children. Type 1 cells in the human thymus were not CGRP positive, but as in the rat, there were similar TH positive cells in the medulla. It was concluded that in addition to nerves containing CGRP, noradrenaline or dopamine, epithelial cells also contain these transmitters. They could therefore act on different cells (compared with neural targets) in a paracrine manner.

In 1813 Gantzer described 2 accessory muscles in the human forearm which bear his name (Wood, 1868; Macalister, 1875; Testut, 1884; Le Double, 1897). The more frequent of the 2 accessory muscles or ‘accessorius ad pollicem’ was found to arise from the coronoid process of the ulna, coursing distally to attach into the flexor pollicis longus muscle (flexor pollicis longus accessory head, FPLah). The less frequently observed or ‘accessorius ad flexorem profundum digitorum’ was again found to arise from the coronoid process and course to join into the flexor digitorum profundus (flexor digitorum profundus accessory head, FDPah). Since their initial description, they have been examined in further detail by a number of authors (Wood, 1868; Macalister, 1875; Le Double, 1897; Dykes & Anson, 1944; Mangini, 1960; Malhotra et al. 1982; Dellon & McKinnon, 1987; Kida, 1988). These studies, most of them focusing on the FPLah, all show different results of prevalence, origin, insertion, relations and nerve supply. We undertook this study with the aim of providing a more accurate account of the detailed morphology of both accessory muscles because of the above-mentioned inconsistent anatomical descriptions and the lack of information as to important aspects such as vascular supply, morphology (shape and length) and the coexistence of both accessory heads.

Compared with other joints, the ligaments of the ankle have not been studied in great detail; consequently relatively little literature exists. The positions of the 3 major bands of the lateral collateral ligament are well known and documented (Schafer et al. 1915; Sarrafian, 1983; McMinn, 1994; Palastanga et al. 1994; Williams et al. 1995). The detailed anatomy of the ligaments is, however, relatively complex with variations of the major bands and several minor additional bands being reported (Sarrafian, 1993; Burks & Morgan, 1994; Rosenberg et al. 1995).

Anomalies of the brachial plexus and its terminal branches are not uncommon. Variations in the course and branches of the musculocutaneous nerve have been noted (Clemente, 1985; Bergman et al. 1988) and its absence was reported by Le Minor (1990). Several anomalies were present in the left plexus of a 59-y-old Japanese man (Fig.). There were no anterior and posterior divisions of the middle trunk, although there were communications between the posterior, medial and lateral cords. The musculocutaneous nerve was absent (Le Minor, 1990) and the medial and lateral roots of the median nerve did not unite in the axillary fossa but in the upper arm about 5 cm distal to the lower border of latissimus dorsi (Adachi, 1928; Buch-Hansen, 1955). The hitherto unreported findings were branches arising directly from the lateral cord to supply coracobrachialis, both heads of biceps brachii and brachialis. The lateral cutaneous nerve of the forearm was derived from the lateral cord with a small contribution from the medial root of the median nerve. Since there were communications between the posterior cord (a continuation of the middle trunk) and the medial and lateral cords, it is theoretically possibly, but not proven, that the root values of branches innervating the flexor muscles of the arm and forearm and the skin of lateral border of the forearm were normal.

The embryology and development of the hip joint are complex. In few areas of the body can so many morphological variations be seen. Minor anatomical abnormalities in the acetabular and femoral head shape, femoral–neck angle, and joint congruency, are frequent. Controversies still exist on the importance of these variations and on whether or not they put the patient at risk of developing early degenerative changes (Wedge et al. 1990). Even in clinically asymptomatic hips, the acetabulum is not always of the same shape or same depth. Minor degrees of ‘dysplasia’ have no more effect than to create a shallow acetabulum or possibly predispose to a labral tear (Dorrel & Caterall, 1986). Greater degrees may cause congenital subluxation or dislocation (Wiberg, 1939; Ponseti, 1978a).

Anomalous muscles in the first flexor compartment of the forearm are quite common. Flexor digitorum superficialis (FDS) is a well known example (Kernohan et al. 1984; Ametewee et al. 1985; Elias et al. 1985; Tonkin & Lister, 1985). However, the anatomical variations of muscles, for example flexor digitorum profundus (FDP) in the second flexor compartment, are not rare (Hollinshead, 1969; Williams & Warwick, 1980; Honing et al. 1995; Williams et al. 1995). These muscles are usually asymptomatic and discovered incidentally during anatomical dissection or at operation. However, they may be symptomatic, presenting as an enlarging tumour-like mass simulating a ganglion (Vichare, 1970; Hayes, 1974; Murakami & Todani, 1982) or may cause a compression neuropathy (Hutton & Kernohan, 1981; Simodynes & Cochran, 1981).

During archaeological excavations in the early modern cemetery in Kernavé, Lithuania, a complete skeleton of a presumed adult male individual was found (grave 108). This skeleton showed a short right humerus and missing radius, ulna and hand. Other parts of the skeleton appeared to be normal, characteristic of a robust constitution. The skeletal material was analysed by macroscopic and radiological techniques. Sex and age were determined following the suggestions of the European Association of Anthropologists (Ferembach et al. 1980), measurements were recorded according to Martin (1928) and Bräuer (1988), and the pathological alterations according to Schultz (1988). The robustness and the measurements indicate a male individual, whose age was put at 40–45 y using the combined method (cf. Ferembach et al. 1980; Szilvássy, 1988) of cranial suture closure, spongiosa structure of the proximal humerus and femur and structure of the pubic symphysis. Skeletal elements analysed included both humeri, clavicles and scapulae.

Studying the shoulder joint in a palaeo-osteological context in recent years, I have been struck, like Prescher & Klümpen (1997), by the lack of information about the glenoid notch provided by the standard anatomical texts; indeed equally unexplained is the pear-shape of the glenoid cavity itself.