Forced locomotor activity improves walking ability of male turkeys and modifies carcass characteristics.

1. This trial investigated the effect of forced locomotor activity (training) on walking ability, leg posture, and growth performance, as well as carcass composition and meat quality in male fattening turkeys.2. A flock of 744 commercial turkeys was divided into three experimental groups, either without any training period (control), training from weeks 2 to 8 (short-term), or training from weeks 2 to 21 (long-term). All birds were slaughtered at an age of 21 weeks. To study the effect of short vs. long time lapse between shackling and stunning (suspension time), each training group was split into two halves that were hooked on the shackle for either 15 s or 3 min prior to stunning.3. Long-term physical training, compared to short-term or no training, resulted in better walking ability and in a lower percentage of leg malposition, as assessed at the end of fattening. No effect on final body weight was detected.4. In a subsample, the composition of 80 carcasses was determined by dissection. Long-term training favoured the percentage of the drumstick over that of the breast cut in comparison to the group without any training.5. Meat quality parameters were determined for breast muscle. The pH values 20 min post-mortem were reduced by long-term training, and the highest value was observed for a combination of no training with short suspension. After 24 h, pH values did not differ between experimental groups. The breast muscles were characterised as fast-glycolysing. Prolonged suspension time resulted in higher electrical conductivity after 24 h, and in higher a* values (redness).6. In conclusion, the study revealed that a long-term training period improved walking ability and leg posture of heavy male turkeys. Thus, training can contribute to the improvement of animal welfare in turkey husbandry. Meat quality variables of breast muscles were partially influenced by locomotor activity and suspension time.

Neural histology and neurogenesis of the human fetal and infant brain.

The human brain develops slowly and over a long period of time which lasts for almost three decades. This enables good spatio-temporal resolution of histogenetic and neurogenetic events as well as an appropriate and clinically relevant timing of these events. In order to successfully apply in vivo neuroimaging data, in analyzing both the normal brain development and the neurodevelopmental origin of major neurological and mental disorders, it is important to correlate these neuroimaging data with the existing data on morphogenetic, histogenetic and neurogenetic events. Furthermore, when performing such correlation, the genetic, genomic, and molecular biology data on phenotypic specification of developing brain regions, areas and neurons should also be included. In this review, we focus on early developmental periods (form 8 postconceptional weeks to the second postnatal year) and describe the microstructural organization and neural circuitry elements of the fetal and early postnatal human cerebrum.

Prominent periventricular fiber system related to ganglionic eminence and striatum in the human fetal cerebrum.

Periventricular pathway (PVP) system of the developing human cerebrum is situated medial to the intermediate zone in the close proximity to proliferative cell compartments. In order to elucidate chemical properties and developing trajectories of the PVP we used DTI in combination with acetylcholinesterase histochemistry, SNAP-25 immunocytochemistry and axonal cytoskeletal markers (SMI312, MAP1b) immunocytochemistry on postmortem paraformaldehyde-fixed brains of 30 human fetuses ranging in age from 10 to 38 postconceptional weeks (PCW), 2 infants (age 1-3 months) and 1 adult brain. The PVP appears in the early fetal period (10-13 PCW) as two defined fibre bundles: the corpus callosum (CC) and the fetal fronto-occipital fascicle (FOF). In the midfetal period (15-18 PCW), all four components of the PVP can be identified: (1) the CC, which at rostral levels forms a voluminous callosal plate; (2) the FOF, with SNAP-25-positive fibers; (3) the fronto-pontine pathway (FPP) which for a short distance runs within the PVP; and (4) the subcallosal fascicle of Muratoff (SFM) which contains cortico-caudate projections. The PVPs are situated medial to the internal capsule at the level of the cortico-striatal junction; they remain prominent during the late fetal and early preterm period (19-28 PCW) and represent a portion of the wider periventricular crossroad of growing associative, callosal and projection pathways. In the perinatal period, the PVPs change their topographical relationships, decrease in size and the FOF looses its SNAP-25-reactivity. In conclusion, the hitherto undescribed PVP of the human fetal cerebrum contains forerunners of adult associative and projection pathways. Its transient chemical properties and relative exuberance suggest that the PVP may exert influence on the development of cortical connectivity (intermediate targeting) and other neurogenetic events such as neuronal proliferation. The PVP's topographical position also indicates that it is a major site of vulnerability in hypoxic-ischaemic perinatal brain injury.

POMT1-associated walker-warburg syndrome: a disorder of dendritic development of neocortical neurons.

We have analyzed the morphology and dendritic development of neocortical neurons in a 2.5-month-old infant with Walker-Warburg syndrome homozygotic for a novel POMT1 gene mutation, by Golgi methods. We found that pyramidal neurons frequently displayed abnormal (oblique, horizontal, or inverted) orientation. A novel finding of this study is that members of the same population of pyramidal neurons display different stages of development of their dendritic arborizations: some neurons had poorly developed dendrites and thus resembled pyramidal neurons of the late fetal cortex; for some neurons, the level of differentiation corresponded to that in the newborn cortex; finally, some neurons had quite elaborate dendritic trees as expected for the cortex of 2.5-month-old infant. In addition, apical dendrites of many pyramidal neurons were conspiciously bent to one side, irrespective to the general orientation of the pyramidal neuron. These findings suggest that Walker-Warburg lissencephaly is characterized by two hitherto unnoticed pathogenetic changes in the cerebral cortex: (a) heterochronic decoupling of dendritic maturation within the same neuronal population (with some members significantly lagging behind the normal maturational schedule) and (b) anisotropically distorted shaping of dendritic trees, probably caused by patchy displacement of molecular guidance cues for dendrites in the malformed cortex.

The morphological detection of bone and cartilage particles in mechanically separated meat.

The calcium content of mechanically separated meat (MSM) is a common indicator of elevated bone contents due to the separation process, e.g. as applied in EU regulations. But a direct morphological proof of bone material is needed, especially to identify undeclared MSM additives to meat products. We present a new morphological method for the detection of both bone and cartilage particles, because elevated cartilage contents may be indicative of MSM additives also. The method is based on simultaneous staining with Alizarin Red and Alcian Blue. We investigated MSM, commercially produced from breast of veal, for elevated particle contents. In addition, we determined the rate of recovery for minced pork mixed with 0-5% bone or cartilage. The results show that staining with Alizarin Red and Alcian Blue is suited to the efficient and simultaneous detection of bone and cartilage material in meat mixtures. The morphological staining is simpler than previously applied methods. Preparations can be assessed macroscopically, and they can be quantified gravimetrically. Manual sorting and weighing of particles appears to underestimate MSM contents if particles are ground very finely.

Nitrinergic neurons in the developing and adult human telencephalon: transient and permanent patterns of expression in comparison to other mammals.

A subpopulation of cerebral cortical neurons constitutively express nitric oxide synthase (NOS) and, upon demand, produce a novel messenger molecule nitric oxide (NO) with a variety of proposed roles in the developing, adult, and diseased brain. With respect to the intensity of their histochemical (NADPH-diaphorase histochemistry) and immunocytochemical (nNOS and eNOS immunocytochemistry) staining, these nitrinergic neurons are generally divided in type I and type II cells. Type I cells are usually large, intensely stained interneurons, scattered throughout all cortical layers; they frequently co-express GABA, neuropeptide Y, and somatostatin, but rarely contain calcium-binding proteins. Type II cells are small and lightly to moderately stained, about 20-fold more numerous than type I cells, located exclusively in supragranular layers, and found almost exclusively in the primate and human brain. In the developing cerebral cortex, nitrinergic neurons are among the earliest differentiating neurons, mostly because the dominant population of prenatal nitrinergic neurons are specific fetal subplate and Cajal-Retzius cells, which are the earliest generated neurons of the cortical anlage. However, at least in the human brain, a subpopulation of principal (pyramidal) cortical neurons transiently express NOS proteins in a regionally specific manner. In fact, transient overexpression of NOS-activity is a well-documented phenomenon in the developing mammalian cerebral cortex, suggesting that nitric oxide plays a significant role in the establishment and refinement of the cortical synaptic circuitry. Nitrinergic neurons are also present in human fetal basal forebrain and basal ganglia from 15 weeks of gestation onwards, thus being among the first chemically differentiated neurons within these brain regions. Finally, a subpopulation of human dorsal pallidal neurons transiently express NADPH-diaphorase activity during midgestation.

Croatian institute for brain research: bridging the gap from the decade of the brain to the next millenium.

The Croatian Institute for Brain Research has been founded at the Zagreb School of Medicine as a central institution for neuroscience research in Croatia. The major research program of the Institute is based on multidisciplinary research using a variety of approaches, from molecular to clinical, for studying the neurobiological basis of normal and disturbed development and functioning of the human brain. Basic organizational features and social goals of the Institute are: permanent correlation of basic and clinical neuroscience, optimal exploitation of resources and manpower, providing the best education and research opportunities for young investigators in neuroscience, and involvement of Croatian neuroscientists into global professional organizations and research programs. Thus, with the establishment of this Institute, Croatia strives to contribute to the overall advancement of neuroscience in The Decade of the Brain and to take part in pushing the frontiers of research along the path leading into the next millenium.

Transient patterns of organization of the human fetal brain.

Fetal development of the human brain is characterized by continuous transformations and reorganization of the fetal telencephalic wall which consists of transient, cytoarchitectonically defined cellular compartments, the so-called embryonic/fetal zones. The cellular and fiber content of these zones is permanently changing, so that fetal neuronal circuitry elements (afferent fibers, synapses, and postsynaptic neurons) display transient patterns of areal, laminar, and modular organization. In the late human fetus and preterm infant, transient patterns of structural and physiological organization form the basis of transient behavioral states and patterns of activity. The transient subplate zone is a key compartment for transient fetal neuronal circuitry, and competitive cellular interactions within the subplate zone are crucial for the areal specification of the cerebral cortex and the formation of cortical connectivity. The subplate zone may also have a key role in cortical repair and plasticity after perinatal brain lesions.

Transformation of the shape of the ethmoid bone during the course of life.

The ethmoid bone undergoes significant changes in the course of life. The critical period is the time from 9 to 35 years of age. The changes in its shape depend upon the intensity of pneumatization of the ethmoid labyrinth and frontal sinus. If pneumatization of the labyrinth is excessive, the ethmoidal cells are not covered by the frontal bone. On the contrary, they are incorporated in the inferior floor of the anterior cranial fossa, resulting in a possible risk of penetration during surgical ethmoidectomy.

Anti-GM3 (II3Neu5Ac-lactosylceramide) ganglioside antibody labels human fetal Purkinje neurons during the critical stage of cerebellar development.

The ganglioside GM3 (II3Neu5Ac-lactosylceramide) represents a minor ganglioside in normal human brain compared to major gangliosides with gangliotetraose-backbone. In this study the presence of GM3 in three 23 and 24 weeks of gestation old human cerebella was demonstrated by immunostaining extracted gangliosides on thin-layer chromatography plate as well as by immunohistochemical analysis of cerebellar cryosections. During this stage of brain development GM3 was found to be dominantly expressed on cells corresponding to Purkinje neurons. Delipidation of histological sections with chloroform/methanol caused significant reduction of anti-GM3 immunostaining, thus confirming the prevalent ganglioside nature of this antigen. These results give evidence that (1) GM3 ganglioside is associated with a specific subset of human fetal cerebellar neurons during the critical development stage, and (2) a definite ganglioside in general is distributed to a specific subset of cells in normal human brain.

Ontogenesis of goal-directed behavior: anatomo-functional considerations.

Recent neuroanatomical and neurophysiological studies in man have revealed ontogenetic events which coincide with broadly defined phases of behavioral and cognitive development. During the early fetal period, early produced neurons make initial synapses which form the basis for the earliest electrical activity of the human brain. The overall immaturity of neuronal connections, in particularly in cortical areas, correlates with the absence of any behavioral pattern or goal-directed movements. In the late fetus and preterm infant, transient accumulation of major afferent pathways, the presence of transient layers (subplate zone) and transient pattern of transmitter-related organization form the neurological basis of cortical electric responses as well as transient behavioral states and sleep patterns. Parallel to the profound structural and chemical reorganization of the human cerebrum during the first 6 postnatal months there is a disappearance of transient behavioral and motor patterns. The previously close spatio-temporal correlation between these events becomes progressively looser. The overproduction of circuitry elements during the subsequent period peaks in associative cortex between 1 and 2 years of age, corresponding to the emergence of skilled actions and cognitive functions. After the elimination of some circuitry elements after the second year of life, the prolonged maturation of goal-directed behavior and the protracted emergence of different cognitive functions correlates with the development plateau of synapse production which can be seen up to 16 years of age. Parallel to the prolonged maturation of postsynaptic elements, there are well defined maturational changes in the chemical properties of associative pyramidal neurons of cortical layer III. These findings correspond to the prolonged maturation of movement-related brain macropotentials as well as other cognition-related potentials, where the last prominent changes were seen after 10 years of age. Although the coincidence of the developmental events does not necessarily mean a causal relationship, the combination of structural and physiological data opens new vistas for the further investigation of the neurobiological basis of goal-directed movement and cognitive behavior.

Identification of war victims in Croatia.

The intention of this paper is to describe the organizational principles and indicate the results already achieved in the identification of war victims in Croatia. By 25 February 1993, 6,493 victims had been identified. A model is proposed that could be used in the course of identification processes, examining the methods and principles of identification which have been complicated by the time interval of more than a year from the time of death, for a presumed number of several thousand (up to 14,000) unidentified victims, possibly in mass graves. Identification is further complicated by the lack of ante-mortem medical and dental records and the incapacity to utilize more expensive methods of identification. Attention is drawn to a group of more complex cases examined at the Institute of Forensic Medicine and Criminology.

Early areal differentiation of the human cerebral cortex: entorhinal area.

The early cytoarchitectonic specialization and area-specific differentiation of the prospective entorhinal cortex were studied in the postmortem human fetal brains (9.5-13.5 postovulatory weeks). At 10 weeks, using the Golgi method, we saw the appearance of area-specific large neurons (promoter neurons) with widely bifurcating apical dendrites situated at the outer margin of the cortical plate of the prospective entorhinal cortex. The analysis of the serial Nissl-stained sections revealed the concomitant appearance of a one-cell-thick layer (monolayer) at the interface between the cortical plate and the marginal zone and multilaminated spread of the deep part of the cortical plate. This is the earliest sign of area-specific cytoarchitectonic differentiation of all pallial regions characterized by the presence of the typical cortical plate. The first subareal differentiation within the entorhinal cortex begins at 13 postovulatory weeks with uneven development of fiber-rich lamina dissecans, which separates two cellular laminae principals (externa and interna), and with the appearance of characteristic cell islands of the prospective layer II. At rostral levels, cell islands begin to develop in the rostromedial entorhinal area at the subpial depths where large promoter neurons reside. At intermediate levels, both lamina dissecans and lamina principalis interna are well delineated. At caudal levels, lamina principalis interna is continuous with the upper subplate zone of the adjacent neocortex. Both area-specific neurons (promoters) and fiber-rich (afferent) strata develop synchronously during the earliest areal differentiation of the cerebral cortex. The precocious lamination of the cortical plate is the crucial event in the histogenesis of the entorhinal cortex.

Zagreb research collection of human brains for developmental neurobiologists and clinical neuroscientists.

The aim of this paper was to offer for the first time a selective and systematic description of the "Zabreb Neuroembryological Collection" of human brains and to illustrate the major results of our research team. Throughout these 16 years of continuous and systematic research, we have applied different techniques for demonstrating the cytoarchitectonics (Nissl staining), neuronal morphology (Golgi impregnation), synaptogenesis (EM analysis), growing pathways (acetylcholinesterase histochemistry) and transmitter-related properties of developing neuronal populations (immunocytochemistry and acetylcholinesterase histochemistry) on several hundred human brains ranging in age from the 5th week post-conception to 90 years. The combination of classical and modern research techniques applied to the constantly growing developmental collection, as well as the continuous evaluation of our data in the light of experimental work in non-human primates, has led to the discovery of an early synaptogenesis within the human cortical anlage and hitherto undescribed transient subplate zone; our results also provided the first comprehensive evidence concerning the timing and pattern of development of afferent fiber systems in the human cortex. All this enabled us to offer a well-documented and coherent reconstruction of major histogenetic events in the human brain. We concluded that structural remodeling and reorganization of the brain, from the transient patterns of the fetal organization through the postnatal phase of transient overproduction of circuitry elements to the final maturation, is the crucial principle of development. Fetal neuronal elements (afferents, synapses and postsynaptic neurons) display transient patterns of laminar, vertical and modular organization and transient cellular interactions and competition in the subplate zone are crucial for the formation of cortical connections. The elucidation of the nature and timing of these histogenetic reorganizational events in the human brain represents the first step towards determining the neurobiological basis of the emergence of behavior, neural functions and cognition in human fetuses, infants and children, which takes place during perinatal and early postnatal life.

[Civilian massacre in Banija: Kraljevcani and Pecki, 14-16 March 1991]. / Pokolji civila u Baniji: Kraljevcani i Pecki 14. i 16. kolovoza 1991.

On 14 and 16 August 1991, Serbian terrorists attacked two villages near Petrinja in the Banija region. The village of Kraljevcani was attacked on 14 August, when the terrorists killed five older villagers, 3 women and 2 men who stayed in the village to watch the livestock and the houses. The women were killed by automatic rifle fire and grenades, and the bodies of the two men were blown up by a hand rocket-launcher projectiles. On 16 August 1991, Serbian terrorists captured four villagers from Pecki, who came to the deserted village to feed the livestock left after the inhabitants fled from the terrorists. The forensic medical expertise revealed that they were first wounded by rifle fire, then tortured and finally executed by hand axes and bayonets.

Neuroscience in Yugoslavia.

This survey is a personal account of the present status of neuroscience in Yugoslavia within the context of recent upheavals in Eastern Europe. The current situation in Yugoslavia, characterized by the absence of a Federal Ministry of Science and a poor scientific communication between federal states (republics), does not allow a comprehensive overview of neuroscience at the federal level. Even more difficult is to envisage the prospects of Yugoslav neuroscience in the light of European integration. Several problems serve to illustrate the present situation concerning Yugoslav neuroscience. First, the weakness of the self-organization of science in Yugoslavia during the past 20 years is still the most important denominator in the current trend of neuroscience. Second, different Yugoslav republics have significantly different systems of science funding and evaluation, which reflect very plainly different levels of democratic (and socioeconomic) changes that were attained during 1990. Third, due to the different numbers of trained scientists, facilities and equipment, funds and levels of international scientific cooperation there are major differences between republics in the tempo of progress towards real achievements in science. Finally, the present explosive development of neuroscience and the proclamation of the 'Decade of the Brain' will hopefully stimulate Yugoslav neuroscientists to seek better programmes of neuroscience research and to improve the extent and quality of international cooperation.