Terminology and labeling of cellular products-2: Implementation plan.

The publication of new standards for terminology and labeling marks an important step in ensuring consistency and traceability of cellular therapies at the global level. However, it is only with the widespread implementation of the standard that the benefits can be truly realized. This paper provides guidance on the practical aspects of adopting these new standards for organizations with differing current levels of computerization. It discusses project management, equipment, licensing, and validation topics.

Terminology and labeling of cellular products: 1. Standards.

The International Cellular Therapy Coding and Labeling Advisory Group was established to address the growing need for standardization of terminology and labeling for cellular therapy products as a result of increasing international transfer of these products. This paper presents new standards for terminology and labeling. These standards have been developed through a consultative process and are supported by key professional and accreditation bodies. By using these standards, together with the unique donation identification numbers and international product reference tables provided by the International Society of Blood Transfusion (ISBT) 128 Standard, consistency and traceability can be assured at the global level. A companion paper provides guidance on the implementation of the ISBT 128 system.

ISBT 128 blood labeling: introduction and reference laboratory applications.

ISBT 128 will be implemented in the United States during the next two years. In addition to improving unit traceability and lookback tracking, this information technology standard has the power to detect and prevent errors in data entry by using data identifiers and check characters. Additionally, its ability to encode special testing results such as CMV and RBC phenotype on a label provides laboratories a computerized mechanism to verify the accuracy of such labels.

Synaptic connections between identified neuron types in the antennal lobe glomeruli of the cockroach, Periplaneta americana: II. Local multiglomerular interneurons.

Synapses between three types of antennal lobe neurons, namely, local multiglomerular interneurons, antennal receptor neurons, and gamma-aminobutyric acid (GABA)-immunoreactive neurons, were studied by means of a combination of three different markers. The interneurons were labeled by intracellular injection of horseradish peroxidase (HRP) into a single soma or a small group of neurons. Antennal receptor cells were marked by experimentally induced anterograde degeneration, and GABA-containing neurons were identified by postembedding immunogold staining. The following types of connections were found: Local interneurons receive input synapses from 1) degenerated receptor neuron axons, 2) GABA-immunogold-labeled neurites, and 3) non-GABA-immunoreactive neurons. The interneurons form output synapses onto the same three neuron groups. Contacts were also found between HRP-labeled interneurons themselves. The majority of synapses were dyadic. In most cases, only one postsynaptic neuronal process of the dyads was labeled and, thus, was identified. Polysynaptic connections were found between GABA-immunoreactive neurites, HRP-labeled interneuron processes, and nonlabeled neurites or between HRP-labeled interneuron processes and two interconnected GABA-immunoreactive processes. The present findings provide anatomical evidence for an earlier suggested monosynaptic connection between afferent receptor fibers and local, at that time putative, GABAergic interneurons. They further reveal that local multiglomerular interneurons are synaptically interconnected. The interneurons, in addition, form serial connections via more than one GABA-immunoreactive neuron with non-GABA-immunoreactive and putative projection neurons. Such polysynaptic connections would be a substrate for a feed-forward "disinhibition" of projection neurons, which has been suggested on the basis of electrophysiological findings.

Synaptic connections between identified neuron types in the antennal lobe glomeruli of the cockroach, Periplaneta americana: I. Uniglomerular projection neurons.

A combination of three different labels was used to demonstrate synapses between three types of neurons within the glomeruli: 1) antennal receptor cells, 2) gamma-aminobutyric acid (GABA)-immunoreactive neurons, and 3) uniglomerular projection neurons. Receptor cell axons were experimentally severed and caused to degenerate; uniglomerular projection neurons, a subgroup of glomerular output neurons, were labeled by intracellular horseradish peroxidase (HRP) injection and GABA-containing neurons by postembedding immunogold staining. The following synaptic connections were identified: 1) Receptor cell axons form monosynaptic contacts in a dyadic fashion onto a dendritic process of a uniglomerular projection neuron and in addition onto a GABA-immunoreactive neuron. 2) Receptor cell axons form polysynaptic connections with dendrites of uniglomerular projection neurons via GABA-immunoreactive neurons. 3) GABA-immunoreactive neurons form dyadic output synapses onto receptor cell axons and in addition onto projection neuron dendrites. These findings provide further evidence that signal transfer from receptor cells onto uniglomerular projection neurons is mediated by two different paths: first, a monosynaptic and presumably excitatory route and, second, an inhibitory polysynaptic route via GABAergic, most likely multiglomerular interneurons. The output synapses of GABA-immunoreactive neurons onto both receptor cells and uniglomerular projection neurons are assumed to exert control functions in regulating the neuronal activity within the glomeruli.

Synaptic connection between olfactory receptor cells and uniglomerular projection neurons in the antennal lobe of the American cockroach, Periplaneta americana.

Both antennal receptor cell axons and uniglomerular projection neurons of the antennal lobe were specifically labeled, and their synaptic relationship was studied at the fine structural level. The labelings were applied in different combinations: i) Experimentally induced anterograde degeneration of sensory-afferent axons was combined with injection of horseradish peroxidase into uniglomerular projection neurons. ii) Lucifer Yellow was injected into uniglomerular projection neurons, and receptor cell axons were anterogradely labeled with the lipophilic dye DiI. The fluorescent dyes were transformed by immuno- or photochemical treatment into electron-dense markers. In both types of preparations, a considerable number of monosynaptic output synapses from antennal receptor neurons onto processes of uniglomerular projection neurons were identified within the glomeruli of the lobe. In most cases, the receptor axon was connected in a dyadic fashion firstly to a process of a projection neuron and secondly to a nonlabeled process. The results clearly demonstrate a direct connection between receptor cells and output neurons of the cockroach antennal lobe which exists in parallel to the already proposed and demonstrated polysynaptic connection via inhibitory local interneurons.

An improved model of the synaptic organization of insect olfactory glomeruli.

In order to characterize the neuronal pathways underlying central nervous processing of odor signals, the synaptic circuitry of identified neuron individuals or types within cockroach antennal lobe glomeruli has been studied in the electron microscope. The anatomical findings are consistent with physiological data from other investigations. For the characterization of the synaptic connections, different combinations of neuron labelings have been applied including (a) horseradish peroxidase injection of local interneurons (INs) or uniglomerular projection neurons (uPNs), (b) gamma-aminobutyric acid (GABA)-immunogold labeling, and (c) experimentally induced degeneration of antennal receptor neuron projections (RNs). The following pathways between glomerular input and output neurons are proposed: (1) A monosynaptic excitatory route, and (2) at least two polysynaptic routes, one of them probably inhibitory and the other disinhibitory. (3) Inhibitory feedback connections exist from these two paths via GABA-positive INs onto both RNs as well as uPNs. (4) Since a given GABA-positive IN innervates a large number of antennal lobe glomeruli, inhibitory signals could also be passed to neighbored glomeruli.

Central projections of the maxillary and antennal nerves in the mosquito Aedes aegypti

In the mosquito Aedes aegypti, CO2-sensitive receptor neurones are located together with two other types of chemoreceptor neurones in club-shaped sensilla basiconica on the most distal segment of the maxillary palps. In order to identify the central target neuropiles of these neurones and to determine whether antennal receptor neurones project into the same area, the palpal and antennal nerves were labelled by anterograde staining with horseradish peroxidase and by experimentally induced degeneration. The different methods revealed a consistent projection pattern. (1) Maxillary afferents project into the suboesophageal ganglion and ascend further into the ipsilateral antennal lobe. There, they terminate within an identified glomerulus of the ventroposterior lobe. (2) Afferents of the antennal flagellum project into all glomeruli of the ipsilateral antennal lobe, with the exception of the glomerulus innervated by the maxillary nerve. The present anatomical findings suggest that primary processing of information about CO2 levels takes place in a defined glomerulus which also receives input from other palpal chemoreceptor neurones.

Localization of odor-induced neuronal activity in the antennal lobes of the blowfly Calliphora vicina: a [3H] 2-deoxyglucose labeling study.

The distribution of odor-evoked neuronal activity in the antennal lobes of the blowfly Calliphora vicina has been studied by means of [3H] 2-deoxyglucose (2-DG) uptake. Stimulation with natural attractants like meat or cheese induced uptake of 2-DG in a large number of glomeruli, stimulation with the single odorant butyric acid only in a small number of glomeruli. In control brains, the majority of glomeruli showed low 2-DG uptake. The distribution of labeled glomeruli was odor-specific and consistent in different specimen. The labeling patterns evoked by different odors overlapped partially but were clearly distinct. The highest uptake within the glomeruli was found in the axons of antennal receptor neurons. The present data provide evidence that different odors are represented as defined spatial patterns of activity across the antennal lobe glomeruli. The overlapping patterns suggest that certain glomeruli participate in the nervous processing of different odors.

Development of AChE-positive neuronal projections from basal forebrain to cerebral cortex in organotypic tissue slice cultures.

Development of the innervation of the cerebral cortex by acetylcholinesterase (AChE)-stained basal forebrain neurons was studied in vitro using the roller tube technique. Slice cultures were maintained from 3 days to 4 weeks either in serum based medium or in chemically defined medium, each supplemented in some cases with nerve growth factor (NGF). The distribution of AChE and choline acetyltransferase (CAT)-containing neurons was investigated using histo- and immunocytochemical techniques. Slice cultures of basal forebrain revealed the presence of large and medium sized AChE-positive neurons. Within one week of cultivation, numerous AChE-labeled fibers could be seen growing out from the basal forebrain toward the cortex. After entering cortical tissue most of the afferent basal forebrain fibers projected either radially or obliquely into the cortical layers. Many afferent axons initially also travelled tangentially within the white matter, and turned then to grow into the cortical layers. Cerebral cortex tissue maintained a coarse laminar organization. Ramifications of basal forebrain fibers were visible within the subplate region, the deep and superficial cortical layers, and within the marginal zone; greatest density occurred in the subplate region and in marginal zones. Many of these processes exhibited branching patterns markedly similar to those observed during cortical development in vivo. Cortex slices placed with the pial surface adjacent to the basal forebrain revealed AChE-stained fibers that entered the cortical tissue through the marginal surface and gave off ramifications within the superficial layers and, less frequently, the deeper cortical layers. CAT-immunostaining revealed labeled cell bodies and neurites only in the basal forebrain, not in the cortex tissue. Control experiments with co-cultures of basal forebrain and cerebellum slices showed no AChE-positive fiber ingrowth into the cerebellum tissue. The results of these studies demonstrate that basal forebrain projections to cerebral cortex in vitro appear similar to the projections that develop in vivo, and indicate that organotypic co-cultures provide a valuable model for studies of developing cortical afferents.

Synaptic connections of dopamine-immunoreactive neurons in the antennal lobes of Periplaneta americana. Colocalization with GABA-like immunoreactivity.

Dopamine-like immunoreactivity was demonstrated histochemically in about ten local interneurons in the antennal lobe of Periplaneta americana. The somata of these neurons are within the ventrolateral group of cell bodies. Additional immunohistochemical tests revealed that the same neurons also have a GABA-like immunoreactivity. Immunohistochemical dopamine staining (preembedding) of preparations in which the antennal receptor fibers had been caused to degenerate showed that in the glomerular neuropil these antennal fibers form output synapses on dopamine-immunoreactive neurons. The latter form output synapses on unstained neuron profiles.

GABA-immunohistochemistry as a label for identifying types of local interneurons and their synaptic contacts in the antennal lobes of the American cockroach.

Synaptic contacts between GABA-immunoreactive neurons, antennal receptor fibers and non-GABA-immunoreactive neurons in the glomerular neuropil of the antennal lobes have been identified by means of a combination of (i) immunohistochemical labeling and (ii) labeling of afferent fibers of the antenna by experimentally induced degeneration. Characteristic contacts of these neurons are: a) Serially arranged polysynaptic contacts between degenerated antennal fibers, GABA-immunoreactive neurons and non-GABA-immunoreactive neurons. b) Monosynaptic contacts between degenerated antennal fibers and non-GABA-immunoreactive neurons. c) Reciprocal synaptic contacts between immunostained and non-stained neurons and synaptic contacts between individual GABA-immunoreactive neurons. d) Synaptic output contacts of GABA-immunoreactive neurons with degenerated antennal fibers. GABA-immunoreactive neuron profiles in the glomeruli are assigned to multiglomerular local interneurons (Distler 1989a); non-immunolabeled profiles may be assigned to projection neurons and other not yet identified interneurons.

Histochemical demonstration of GABA-like immunoreactivity in cobalt labeled neuron individuals in the insect olfactory pathway.

Individual olfactory neurons in the antennal lobes of Periplaneta americana were investigated for their morphological and neurochemical properties by means of intracellular cobalt injection and indirect peroxidase-antiperoxidase immunohistochemistry. GABA-like immunoreactivity was demonstrated in many local interneurons but not in uniglomerular projection neurons.

Serotonin-immunoreactive neurons in the antennal lobes of the American cockroach Periplaneta americana: light- and electron-microscopic observations.

A large deutocerebral serotonin-immunoreactive neuron arborizes profusely in the glomeruli of the antennal lobes, and also sends neurites into the lateral lobe and the calyces of the mushroom bodies in the ipsilateral protocerebrum. Electron micrographs of the glomerular neuropil show that the main synapses of the serotonin-immunoreactive arborizations are output contacts with unidentified neuron profiles. Only a few synaptic input contacts with serotonin-labeled fibers were observed.

Are right- and left-sided colon neoplasms distinct tumors?

Cancer of left and right colon has a differing prevalence at varying ages, in high- and low-incidence nations, as well as in men and in women. There also is a difference in clinical presentation, in prognosis, and possibly in genetic and environmental epidemiology. This review proposes that cancers of proximal and distal colon are different tumors because of their embryologic origin, genetic changes, and biologic identity. These factors are important in understanding the 'shift of tumors from more distal to more proximal sites in the colon' and in evaluating potential suggestions for instituting advances in diagnosis and prevention.

Formation of synapses between basal forebrain afferents and cerebral cortex neurons: an electron microscopic study in organotypic slice cultures.

Co-cultures of rat basal forebrain and cerebral cortex were maintained from 1 to 5 weeks in vitro with serum-free defined medium. The formation of synaptic connections between basal forebrain afferent fibres and cortical neurons was studied by specific labelling with three staining techniques, including (i) neuronal tract tracing with the fluorescent dye 1,1'-dioctodecyl-3,3,3'3'- tetramethylindocarbocyanine perchlorate, (ii) acetylcholinesterase histochemistry, and (iii) choline acetyltransferase immunocytochemistry. Both basal forebrain and cerebral cortex tissue displayed organotypic characteristics in culture. Cerebral cortex revealed a dense innervation by axonal projections from the basal forebrain. All three labelling techniques produced similar results at the light microscopic level, with densest innervation located in the marginal zone. At the fine structural level, the 1,1'-dioctodecyl-3,3,3'3'-tetramethylindocarbocyanine perchlorate-, acetylcholinesterase- and choline acetyltransferase-stained basal forebrain afferents all revealed a number of synaptic contacts with cortical neurons. The contacts displayed consistent synaptic features, including presynaptic accumulation of small round vesicles, cleft widening, and postsynaptic densities forming symmetric synapses. These morphological characteristics of connections formed in vitro are similar to basal forebrain cholinergic projections to cerebral cortex in normal brain. Based on these results, this tissue culture model appears to be an useful tool for investigations of the development of cholinergic innervation of cerebral cortex.

Synaptic connections between GABA-immunoreactive neurons and uniglomerular projection neurons within the antennal lobe of the cockroach, Periplaneta americana.

Synapses within deutocerebral glomeruli between GABA-immunoreactive, putatively inhibitory local interneurons and uniglomerular projection (output) neurons were demonstrated by means of a combination of GABA-immunogold labeling and intracellular HRP injection. The following connections were identified. 1) GABA-immunoreactive (GABAir) neurons form output synapses in a dyadic fashion onto a uniglomerular projection neuron and, in addition, a second GABAir neuron. A uniglomerular projection neuron in turn forms dyadic output synapses onto two GABAir neurons. Several examples of reciprocal connections have been identified between, first, GABAir neurons and uniglomerular projection neurons, and, second, GABAir neurons themselves. 2) GABAir neurons are serially connected with uniglomerular projection neurons via interposed GABAir processes. In some cases, also the first GABAir process of such a polysynaptic connection formed an output synapse onto the projection neuron. Such serial connections may form the structural basis for both, the feedforward inhibition as well as the feedforward disinhibition of uniglomerular projection neurons by GABAergic neurons. The reciprocal contacts may serve as control devices that modulate the output activity of the projection neurons.

Regional distribution of carcinogen-induced colonic neoplasia in the rat.

Carcinogen induction of neoplasms in rodent colon has been used as a model for human colon cancer development and for evaluating chemopreventive regimens. We studied the regional distribution of small and large intestinal tumors in 229 rats given azoxymethane (AOM) once weekly for two weeks (15 mg/kg sc). The AOM regimen induced 63% more tumors in distal (DC) than in proximal colon (PC), although tumor volume was greater in PC. A high-fat (23% corn oil) diet increased tumors in PC and DC (p < 0.01). Caloric restriction of 10-30% of the ad libitum diet progressively reduced DC tumor formation but did not alter PC tumors. Tumor volume was unaffected by either regimen. Small intestinal tumors were concentrated in the proximal 15 cm of the intestine and were unaffected by dietary manipulation. This AOM model of colon tumor formation approximates human colon cancer distribution and is an appropriate model for rodent chemopreventive studies.