A histochemical, light and electron microscopy study of the hatching gland cells (HGCs) in incubated 50-d-old trout embryos is reported. The distribution of carbohydrate residues in the glycoconjugates of these cells was studied by means of a battery of 13 different lectins conjugated with horseradish peroxidase (PNA, ConA, LCA, WGA, SBA, UEA-I, HPA, DBA) or digoxigenin (DSA, MAA, AAA, SNA, GNA). Identification of N- and O-linked oligosaccharides in HGCs was performed by application of both chemical and enzymatic treatments. Present results suggest that HGCs are seromucous cells which store both high choriolytic enzyme (HCE) and low choriolytic enzyme (LCE), and that their cytoplasmic granules, endoplasmic reticulum and Golgi complex contain additional sialic acid-rich glycoproteins. The negative charge of these glycoproteins might be responsible for the rapid expansion of mucin to form a highly hydrated gel, which would facilite the action of these enzymes in programmed cell death and might play a major role during the morphogenic events.

We have previously shown that exposing rats to a high dose of ethanol on postnatal d 5 can affect Purkinje cell numbers in the cerebellum whilst similar exposure on d 10 had no such effect. The question arose whether a longer period of ethanol exposure after d 10 could produce loss of Purkinje cells. We have examined this question by exposing young rats to a relatively high dose (∼420–430 mg/dl) of ethanol for 6 d periods between the ages of either 4 and 9 d or 10 and 15 d of age. Exposure was carried out by placing the rats in an ethanol vapour chamber for 3 h per day during the exposure period. Groups of ethanol-treated (ET), separation controls (SC) and mother-reared controls (MRC) were anaesthetised and killed when aged 30 d by perfusion with buffered 2.5% glutaraldehyde. Stereological methods were used to determine the numbers of Purkinje cells in the cerebellum of each rat. MRC, SC and rats treated with ethanol between 10–15 d of age each had, on average, about 254–258 thousand cerebellar Purkinje cells; the differences between these various groups were not statistically significant. However, the rats treated with ethanol vapour between 4–9 d of age had an average of only about 128000±20000 Purkinje cells per cerebellum. This value was significantly different from both the MRC and group-matched SC animals. It is concluded that the period between 4 and 9 d of age is an extremely vulnerable period during which the rat cerebellar Purkinje cells are particularly susceptible to the effects of a high dose of ethanol. However, a similar level and duration of ethanol exposure commencing after 10 d of age has no significant effect on Purkinje cell numbers.

Anatomical Society of Great Britain and Ireland

Future meetings

13–15 July 1999. University of Glasgow. This meeting will feature a symposium on the neurobiology of the basal ganglia (to include basic neuronal properties, disease aspects including imaging and transplantation, and evolutionary aspects).

11–16 September 1999. 15th Congress of the International Federation of Association of Anatomists, Rome, Italy.

17 September 1999. Royal Free and University College Medical School, University of London, Royal Free Campus. One-day symposium on the biology of ageing.

5–7 January 2000. Royal Holloway College, University of London. This will include a symposium on phenotypic changes in epithelial development.

24–26 July 2000. University of Cambridge; this will be a tripartite meeting with the Anatomische Gesellschaft and the Nederlandse Anatomen Vereniging.

The Wellcome Trust

An exhibition entitled The New Anatomists will be held in the Two 10 Gallery at the Wellcome Trust Offices, 210 Euston Road, London NW1 2BE from 11 March to 16 July 1999. It is part of a series exploring the relationship between science and art. The exhibition is open from 9 am to 6 pm on weekdays, public holidays excluded. Admission is free.

Previous quantitative descriptions of cellular ultrastructure have focused on spatial content (volume, surface area and number of organelles and membrane domains). It is possible to complement such descriptions by also quantifying spatial arrangements. Hitherto, applications of stereological methods for achieving this (notably, estimation of covariance and pair correlation functions) have been confined to organ and tissue levels. This study explores 3-dimensional subcellular arrangements of key organelles within acinar cells of rabbit parotid salivary glands, highly polarised epithelial cells specialised for exocrine secretion of α-amylase. It focuses on spatial arrangements of secretion product stores (zymogen granules), rough endoplasmic reticulum (RER) and mitochondria. Systematic random samples of electron microscopical fields of view from 3 rabbits were analysed using test grids bearing linear dipole probes of different sizes. Unbiased estimates of organelle volume densities were obtained by point counting and estimates of covariance and pair correlation functions by dipole counting. Plots of pair correlation functions against dipole length identified spatial arrangement differences between organelle types. Volumes within RER and mitochondrial compartments were positively correlated with themselves at distances below 4 μm and 2 μm respectively but were essentially randomly arranged at longer distances. In sharp contrast, zymogen granules were not randomly arranged. They were clustered at distances below 6–7 μm and more widely scattered at greater distances. These findings provide quantitative confirmation of the polarised arrangement of zymogen granules within acinar cells and further support for the relative invariance of biological organisation between subjects.

We carried out a computerised tomographic (CT) examination to elucidate the modifications in the head related to orbital enlargement in the Baikal seal. Transverse CT images showed that (1) the external frontal contours and the frontal sinuses are compressed medially and ventrally by the orbital enlargement; (2) the caudal part of the nasal cavity and nasopharynx are compacted ventrally; and (3) the cranial cavity is displaced caudally. The neurocranium is obviously separated from the facial part in the transverse plane at the caudal region of the orbit. The disposition of the mandible, zygomatic arch, temporal bone, and the masseter, temporal, digastric and pterygoid muscles is changed by the enlarged orbit in the 3-dimensional reorganisation of the head in this species. It is suggested that adaptation for the Lake Baikal environment primarily resulted in orbital enlargement, and that the altered orbital design may subsequently have influenced the form and function of the masticatory and respiratory system.

Immunohistochemistry of normal eccrine sweat glands was performed on paraffin sections of human skin. Immunoreactivity (ir) for neuron specific enolase, S100 protein (S100), regulatory peptides, nitric oxide synthase type I (NOS-I) and choline-acetyltransferase (ChAT) was found in small nerve bundles close to sweat glands. In the glands, secretory cells were labelled with anticytokeratin antibody. Using antibodies to S100, calcitonin gene-related peptide (CGRP) and substance P (SP) a specific distribution pattern was found in secretory cells. Granulated (dark) and parietal (clear) cells were immunopositive for CGRP, and S100 and SP, respectively. Immunoreactivity was diffuse in the cytoplasm for CGRP and S100, and peripheral for SP. Myoepithelial cells were not labelled. Electron microscopy revealed electron dense granules, probably containing peptide, in granulated cells. Using antibodies to NOS-I and ChAT, ir was exclusively found in myoepithelial cells. Immunoreactivity for the atrial natriuretic peptide was absent in sweat glands. These results provide evidence for the presence of both regulatory peptides involved in vasodilation and key enzymes for the synthesis of nitric oxide and acetylcholine in the secretory coil of human sweat glands. It is suggested that human sweat glands are capable of some intrinsic regulation in addition to that carried out by their nerve supply.

In mdx mice, a model for Duchenne muscular dystrophy, the timing between the replication of myoblasts and their incorporation into myotubes was determined autoradiographically. Thirty-eight mdx mice aged 23 d were injected with tritiated thymidine to label myoblasts replicating early in the dystrophic process. At intervals from 8 h to 30 d after injection the tibialis anterior muscles were removed, processed for autoradiography and analysed for labelled central myonuclei (derived from the progeny of myoblasts which had been labelled at 23 d). At 8 h after injection there were no labelled central myonuclei, showing that the labelled myoblasts had not fused within this time. At 1 d, 2% of central myonuclei were labelled, at 2 d, up to 32% were labelled, at 3 d ∼60% were labelled, and at 4 d the labelling peaked at 74%. In the 27 mice sampled from 5–30 d after injection, the levels of central myonuclear labelling varied enormously: from 1–63%. However, there was a consistent decrease in the numbers of labelled central myonuclei with time. This may have been due to dilution of the relative numbers of labelled myonuclei due to other, nonlabelled, myoblasts replicating after the availability of tritiated thymidine, and fusing. It was also possible that labelled myofibres underwent subsequent necrosis and were eliminated from the muscle. The proposal that a regenerated myofibre can undergo a subsequent cycle of necrosis and regeneration was supported by evidence of some necrotic myofibres with labelled and unlabelled central nuclei. These results have implications for understanding the cellular biology and pathology of dystrophic muscle, particularly in relation to myoblast transfer therapy as a potential treatment of Duchenne muscular dystrophy.

The swelling behaviour of the general matrix of both normal and abnormally softened articular cartilage was investigated in the context of its relationship to the underlying subchondral bone, the articular surface, and with respect to the primary structural directions represented in its strongly anisotropic collagenous architecture. Swelling behaviours were compared by subjecting tissue specimens under different modes of constraint to a high swelling bathing solution of distilled water and comparing structural changes imaged at the macroscopic, microscopic and ultrastructural levels of resolution. Near zero swelling was observed in the isolated normal general matrix with minimal structural change. By contrast the similarly isolated softened general matrix exhibited large-scale swelling in both the transverse and radial directions. This difference in dimensional stability was attributed to fundamentally different levels of fibril interconnectivity between the 2 matrices. A model of structural transformation is proposed to accommodate fibrillar rearrangements associated with the large-scale swelling in the radial and transverse directions in the softened general matrix.

The aim of this study was to characterise pulmonary reimplantation injury in isolated, perfused rat lungs following 2 h of cold ischaemia, and 50 min. of in vitro reperfusion. The effects of 2 differently composed lung preservation solutions (low potassium Euro-Collins and Celsior; each n=5) were examined in comparison with untreated, nonischaemic control lungs (n=3). After fixation by vascular perfusion and tissue collection by systematic random sampling, the volume weighted mean volume (Vv) of alveoli and acinar pathways was estimated by light microscopic stereology using the method of point sampled intercepts in plastic embedded, Azan-stained material. Significantly higher Vv of alveoli and acinar paths was found in the Celsior group than in Euro-Collins preserved lungs. However, in the controls the size of acinar pathways was similar to Celsior preserved lungs whereas alveolar size was comparable to preservation with Euro-Collins. The between-animal coefficient of variation of alveoli was very low in controls and Celsior preserved but higher in the Euro-Collins group. Size distribution of alveoli and acinar paths in 15 size classes was largely homogeneous in all groups tested. In the Euro-Collins group the fractions of both class 1-alveoli and class 1-acinar paths significantly exceeded those of the other groups. Widely expanded alveoli (size classes 13–15) only occurred after preservation with Celsior whereas wider acinar paths (size class 15) were found in the Celsior group and in the controls. It is concluded that lung preservation with low-potassium Euro-Collins and Celsior solutions may act differently on distinct spaces in the distal gas-exchange regions of lungs. This may be due to selective effects on pulmonary surfactant activity and on elastic tissue elements in the alveolar ducts, respectively. Additionally, the method of point sampled intercepts is considered to be an efficient tool to evaluate the effects of different preservation solutions on lung parenchyma.

The morphology and distribution of granulated metrial gland cells and of interstitial trophoblast cells in the uterine wall was studied in the first half of pregnancy in the bank vole, Clethrionomys glareolus. The morphology and distribution of granulated metrial gland cells was generally similar to that found in other members of the Rodentia, although they were absent from the walls of the arterial vessels passing through the decidua basalis. Interstitial trophoblast invaded the decidualising endometrium mesometrial to, and antimesometrial to, the implanted embryos. There was no apparent spatiotemporal relationship between the distribution of granulated metrial gland cells and interstitial trophoblast cells.

This paper presents the dynamic injury tolerances for the female humerus and forearm derived from dynamic 3-point bending tests using 22 female cadaver upper extremities. Twelve female humeri were tested at an average strain rate of 3.7±1.3%/s. The strain rates were chosen to be representative of those observed during upper extremity interaction with frontal and side airbags. The average moment to failure when mass scaled for the 5th centile female was 128±19 Nm. Using data from the in situ strain gauges during the drop tests and geometric properties obtained from pretest CT scans, an average dynamic elastic modulus for the female humerus was found to be 24.4±3.9 GPa. The injury tolerance for the forearm was determined from 10 female forearms tested at an average strain rate of 3.94±2.0%/s. Using 3 matched forearm pairs, it was determined that the forearm is 21% stronger in the supinated position (92±5 Nm) versus the pronated position (75±7 Nm). Two distinct fracture patterns were seen for the pronated and supinated groups. In the supinated position the average difference in fracture time between the radius and ulna was a negligible 0.4±0.3 ms. However, the pronated tests yielded an average difference in fracture time of 3.6±1.2 ms, with the ulna breaking before the radius in every test. This trend implies that in the pronated position, the ulna and radius are loaded independently, while in the supinated position the ulna and radius are loaded together as a combined structure. To produce a conservative injury criterion, a total of 7 female forearms were tested in the pronated position, which resulted in the forearm injury criterion of 58±12 Nm when scaled for the 5th centile female. It is anticipated that these data will provide injury reference values for the female forearm during driver air bag loading, and the female humerus during side air bag loading.

A band of fascial thickening, termed the iliopubic tract, lies on the posterior aspect of the inguinal region and has been described in the surgical literature as playing an important role during herniorraphy. This study was undertaken to examine the gross and microscopic anatomy of the iliopubic tract in 12 cadavers. The results confirmed that the iliopubic tract can be readily identified as a thickening of the transversalis fascia running deep and parallel to the inguinal ligament. It attaches to the superomedial part of the pubic bone medially, but laterally its fibres fan out within the fascia transversalis and fascia iliaca without bony attachment to the iliac spines. In contrast to the inguinal ligament, the histological analysis of the iliopubic tract shows a high elastin to collagen ratio. The functional signficance of this structure merits further study, but there is no doubt that it is important in many approaches to inguinal herniorraphy. For this reason it is considered that the iliopubic tract deserves greater emphasis in the anatomy teaching of the inguinal region.

The anatomy of the psoas major muscle (PMA) in young black and white men was studied during routine autopsies. The forensic autopsies included 44 fresh male cadavers (21 black, 23 white) with an age span of 14 to 25 y. The range for weight was 66–76 kg and for height 169–182 cm. The PMA was initially measured in its entire length before measuring the diameter and circumference at each segmental level (L1–S1). At each segmental level, the calculated anatomical cross-sectional area (ACSA) was more than 3 times greater in the black group compared with the white (P<0.001). The psoas minor muscle (PMI) was absent in 91% of the black subjects, but only in 13% of the white subjects. These data show that the PMA is markedly larger in black than white subjects. The marked race specific difference in the size of the PMA may have implications for hip flexor strength, spine function and race specific incidence in low back pathology, and warrants further investigation.

The most frequent disposition of the structures within the ulnar tunnel is for the ulnar nerve to be located medial or posteromedial to the ulnar artery. The structures within the ulnar tunnel are closely related to the medial part of the flexor retinaculum. Lesions of the ulnar nerve and artery during endoscopic decompression of the carpal tunnel have been reported (Agee et al. 1992; Lee et al. 1992; Nath et al. 1993; De Smets & Fabry, 1995). An adequate anatomical knowledge of such structures and their variations is therefore important. During a study of the palmar region, we found that a special branch originated from the ulnar nerve in relation to the flexor retinaculum. Documentation of this variation will contribute to the knowledge of the anatomy of the ulnar nerve and its distal branches.

Several experimental models have demonstrated that the central nervous system is functionally linked to the immune system by means of the autonomic nervous system. Samples of 36 lymph nodes of patients whose ages ranged from 16 to 69 y were studied. In order to demonstrate the existence and distribution of sympathetic nerve fibres, a polyclonal antibody antityrosine hydroxylase (TH), with the streptavidin-biotin system of detection, was used. TH-positive nerve fibres appeared in all reactive patterns of the lymph nodes studied. Thin nerve fascicles ramified at the hilar region and also in the connective tissue septae. Adventitial adrenergic nerve fibres were found following afferent, and to a lesser extent, efferent blood vessels. Another source of incoming nerve fibres was found at capsular level, accompanying blood vessels. On the arterial side, the innervation ceased before reaching the follicular arterioles. Our demonstration of innervation in postcapillary venules could support a regulatory role of adrenergic neurotransmitters in lymphocyte traffic. Occasional nerve fibres were also seen in T areas among parenchymatous cells. These findings confirm the existence of sympathetic innervation in human lymph nodes, and provide indirect evidence that the psychoneuroimmune axis could also exist in humans.

Neurotisation involves transfer of nerves for the restoration of function following injury. A number of nerves have been used in different part of the peripheral nervous system. This study was undertaken to develop a practical and relatively safe surgical approach to the treatment of L4 root lesion's. We examined the effectiveness and safety of neurotisation of the deep peroneal nerve and its branches by the superficial peroneal nerve. Twelve legs of dissected cadavers provided for teaching purposes in the anatomy laboratory were used to display the common peroneal nerve and its branches. Each branch was measured using calipers and analysed to investigate the possibility of neurotisation of the deep peroneal nerve by the superficial peroneal nerve and its branches. It was found that of the measured branches, transposition was possible between those to peroneus longus and tibialis anterior on the basis of their diameter and length. In recent decades, advances in microsurgical reconstruction and understanding of the microanatomy have played major roles in improving the results of surgical treatment of nerve injuries. There is a need for further experimental studies on the feasibility of this surgical approach.

The gross anatomical features of human cervical vertebrae during the fetal-neonatal period were investigated in order to develop morphological standards for the individual ossification centres for use in forensic and anthropological osteology. It was found that the morphology of the cervical vertebral arches and the centra cannot be used for the determination of fetal age although the dens of the axis displays some developmental differences which may be useful for the determination of fetal maturity.

A superficial artery may be present in the forearm, arising from the axillary, brachial or superficial brachial arteries and crossing over the origin of the flexor muscles of the forearm to reach the palm (Adachi, 1928; Bergman et al. 1988). When this superficial artery continues as the normal ulnar artery accompanying the ulnar nerve at the wrist, it is referred to as the superficial ulnar artery, with an incidence of ∼4%. When the artery passes below or superficial to the flexor retinaculum in the middle of the forearm, sometimes continuing to join the superficial palmar arch, it is called the superficial median artery, with an incidence of ∼1%. We have observed a relatively rare variation involving the presence of a superficial median artery in both upper limbs. We discuss the clinical importance and the developmental aspects of this arterial variation.

The chondroepitrochlearis is an extremely rare muscular anomaly. Bergman et al. (1988) in their book on anatomical variations refer to ‘costoepitrochlearis, chondroepitrochlearis, or chondrohumeralis’ and describe the anomaly as a muscular slip which arises from one or more ribs, crosses the axilla, and inserts into the median intermuscular septum or medial humeral epicondyle. The phylogenetic significance of the muscle has received attention (Landry, 1958; Chiba et al. 1983; Bergman, 1991), and case reports with ulnar nerve entrapment and restriction of arm movements (FitzGerald, 1935–1936; Voto & Weiner 1987; Lin, 1988; Spinner et al. 1991) have emphasised its clinical relevance.