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The axotomy-inducible enzyme Nna1 defines a subfamily of M14 metallocarboxypeptidases, and its mutation underlies the Purkinje cell degeneration (pcd) mouse. However, the relationship among its catalytic activity, substrate specificities, and the critical processes of neurodegeneration/axon regeneration is incompletely understood. Here we used a transgenic rescue strategy targeting expression of modified forms of Nna1 to Purkinje cells in pcd mice to determine structure-activity relationships for neuronal survival and in parallel characterized the enzymatic properties of purified recombinant Nna1. The Nna1 subfamily uniquely shares conserved substrate-determining residues with aspartoacylase that, when mutated, cause Canavan disease. Homologous mutations (D1007E and R1078E) inactivate Nna1 in vivo, as does mutation of its catalytic glutamate (E1094A), which implies that metabolism of acidic substrates is essential for neuronal survival. Consistent with reports that Nna1 is a tubulin glutamylase, recombinant Nna1—but not the catalytic mutants—removes glutamate from tubulin. Recombinant Nna1 metabolizes synthetic substrates with 2 or more C-terminal glutamate (but not aspartate) residues (Vmax for 3 glutamates is ∼7-fold higher than 2 glutamates although KM is similar). Catalysis is not ATP/GTP dependent, and mutating the ATP/GTP binding site of Nna1 has no effect in vivo. Nna1 is a monomeric enzyme essential for neuronal survival through hydrolysis of polyglutamate-containing substrates.—Wu, H.-Y., Wang, T., Li, L., Correia, K., Morgan, J. I. A structural and functional analysis of Nna1 in Purkinje cell degeneration (pcd) mice.

Purkinje cell protein 4-like 1 (Pcp4l1) is a small neuronal IQ motif protein closely related to the calmodulin-binding protein Pcp4/PEP-19. PEP-19 interacts with calmodulin via its IQ motif to inhibit calmodulin-dependent enzymes and we hypothesized Pcp4l1 would have similar properties. Surprisingly, full-length Pcp4l1 does not interact with calmodulin in yeast two-hybrid or pulldown experiments yet a synthetic peptide constituting only the IQ motif of Pcp4l1 binds calmodulin and inhibits calmodulin-dependent kinase II. A nine-residue glutamic acid-rich sequence in Pcp4l1 confers these unexpected properties. This element lies outside the IQ motif and its deletion or exchange with the homologous region of PEP-19 restores calmodulin binding. Conversion of a single isoleucine (Ile36) within this motif to phenylalanine, the residue present in PEP-19, imparts calmodulin binding onto Pcp4l1. Moreover, only aromatic amino acid substitutions at position 36 in Pcp4l1 allow binding. Thus, despite their sequence similarities PEP-19 and Pcp4l1 have distinct properties with the latter harboring an element that can functionally suppress an IQ motif. We speculate Pcp4l1 may be a latent calmodulin inhibitor regulated by post-translational modification and/or co-factor interactions.

J. Neurochem. (2012) 121, 717–729. Abstract Cerebellin precursor protein (Cbln1) is essential for synapse integrity in cerebellum through assembly into complexes that bridge pre-synaptic β-neurexins (Nrxn) to post-synaptic GluRδ2. However, GluRδ2 is largely cerebellum-specific, yet Cbln1 and its little studied family members, Cbln2 and Cbln4, are expressed throughout brain. Therefore, we investigated whether additional proteins mediate Cbln family actions. Whereas Cbln1 and Cbln2 bound to GluRδ2 and Nrxns1-3, Cbln4 bound weakly or not at all, suggesting it has distinct binding partners. In a candidate receptor-screening assay, Cbln4 (but not Cbln1 or Cbln2) bound selectively to the netrin receptor, (deleted in colorectal cancer (DCC) in a netrin-displaceable fashion. To determine whether Cbln4 had a netrin-like function, Cbln4-null mice were generated. Cbln4-null mice did not phenocopy netrin-null mice. Cbln1 and Cbln4 were likely co-localized in neurons thought to be responsible for synaptic changes in striatum of Cbln1-null mice. Furthermore, complexes containing Cbln1 and Cbln4 had greatly reduced affinity to DCC but increased affinity to Nrxns, suggesting a functional interaction. However, Cbln4-null mice lacked the striatal synaptic changes seen in Cbln null mice. Thus, Cbln family members interact with multiple receptors/signaling pathways in a subunit composition-dependent manner and have independent functions with Cbln4 potentially involved in the less well-characterized role of netrin/DCC in adult brain.

Cerebellin precursor protein 1 (Cbln1) is a secreted glycoprotein (Urade et al. 1991; Bao et al. 2005) essential for normal synaptic structure and function in the cerebellum (Hirai et al. 2005). In cerebellum Cbln1 acts as a bi-functional ligand bridging pre-synaptic β-neurexins on granule neurons to post-synaptic Grid2 in Purkinje cells (Uemura et al. 2010; Matsuda et al. 2010). However, this cannot be the sole mechanism of action of Cbln1 as it is expressed in neurons in many brain regions (Mugnaini et al. 1987; Miura et al. 2006; Wei et al. 2007) whereas Grid2 expression is largely confined to cerebellum (Araki et al. 1993). In particular, Cbln1 deficiency in glutamatergic neurons of the parafascicular nucleus (PF) of the thalamus results in increased synaptic spine density on dendrites of striatal medium spiny neurons (MSN), the recipients of axonal projections from the Cbln1-positive PF neurons (Kusnoor et al. 2010). As MSNs and PF neurons lack Grid2 this implies that additional membrane proteins mediate the function of Cbln1 in the thalamo-striatal projection. The dissection of the mechanisms underlying Cbln1 function is further complicated by the fact that Cbln1 is also the prototype of a family of structurally related proteins (Cbln1-4) of largely unknown function (Bao et al. 2005). These proteins have broad, sometimes overlapping, distributions in brain (Miura et al. 2006; Wei et al. 2007) and notably in situ hybridization indicates that Cbln1 and Cbln2 are both expressed in PF neurons (Miura et al. 2006). Besides having the potential to assemble into homomeric complexes all Cbln1 family members are capable of assembling into heteromeric complexes with each other (Bao et al. 2005). This might provide a mechanism to regulate or modify receptor binding specificities, as well as protein turnover, trafficking or function of Cbln1 family members in vivo. For example, Cbln1 and Cbln3 are co-expressed in cerebellar granule cells and are found in heteromeric complexes in vivo (Bao et al. 2006). Moreover, genetic elimination of either Cbln1 or Cbln3 has profound effects on the turnover of the other family member (Bao et al. 2006). Therefore, it is important to determine whether family members have redundant function and underpin similar aspects of neurobiology. As Cbln1 and Cbln2 have broad expression in the adult nervous system, are able to associate with each other in heteromeric complexes, and are co-expressed in PF neurons we have used transgenic and knockout mouse strategies to determine whether they have identical or independent functions in vivo.

Cbln1 (a.k.a. precerebellin) is secreted from cerebellar granule cells as homohexamer or in heteromeric complexes with Cbln3. Cbln1 plays crucial roles in regulating morphological integrity of parallel fiber (PF)-Purkinje cell (PC) synapses and synaptic plasticity. Cbln1-knockout mice display severe cerebellar phenotypes that are essentially indistinguishable from those in glutamate receptor GluRdelta2-null mice, and include severe reduction in the number of PF-PC synapses and loss of long-term depression of synaptic transmission. To understand better the relationship between Cbln1, Cbln3 and GluRdelta2, we performed light and electron microscopic immunohistochemical analyses using highly specific antibodies and antigen-exposing methods, i.e. pepsin pretreatment for light microscopy and postembedding immunogold for electron microscopy. In conventional immunohistochemistry, Cbln1 was preferentially associated with non-terminal portions of PF axons in the molecular layer but rarely overlapped with Cbln3. In contrast, antigen-exposing methods not only greatly intensified Cbln1 immunoreactivity in the molecular layer, but also revealed its high accumulation in the synaptic cleft of PF-PC synapses. No such synaptic accumulation was evident at other PC synapses. Furthermore, Cbln1 now came to overlap almost completely with Cbln3 and GluRdelta2 at PF-PC synapses. Therefore, the convergence of all three molecules provides the anatomical basis for a common signaling pathway regulating circuit development and synaptic plasticity in the cerebellum.

Cbln1 is a secreted glycoprotein essential for synapse structure and function in cerebellum that is also expressed in extracerebellar structures where its function is unknown. Furthermore, Cbln1 assembles into homomeric complexes and heteromeric complexes with three family members (Cbln2-Cbln4), thereby influencing each other's degradation and secretion. Therefore, to understand its function, it is essential to establish the location of Cbln1 relative to other family members. The localization of Cbln1 in brain was determined using immunohistochemistry and cbln1-lacZ transgenic mice. Cbln1-like immunoreactivity (CLI) was always punctate and localized to the cytoplasm of neurons. The punctate CLI colocalized with cathepsin D, a lysosomal marker, but not with markers of endoplasmic reticulum or Golgi, indicating that Cbln1 is present in neuronal endosomes/lysosomes. This may represent the cellular mechanism underlying the regulated degradation of Cbln1 observed in vivo. Outside the cerebellum, CLI mapped to multiple brain regions that were frequently synaptically interconnected, warranting their analysis in cbln1-null mice. Furthermore, whereas CLI increased dramatically in the cerebellum of cbln3-null mice it was unchanged in extracerebellar neurons. This opens the possibility that other family members that are coexpressed in these areas control Cbln1 levels, potentially by modulating processing in the endolysosomal pathway. During development of cbln1-lacZ mice, beta-galactosidase staining was first observed in proliferating granule cell precursors prior to synaptogenesis and thereafter in maturing and adult granule cells. As cbln3 is only expressed in post-mitotic, post-migratory granule cells, Cbln1 homomeric complexes in precursors and Cbln1-Cbln3 heteromeric complexes in mature granule cells may have distinct functions and turnover.

The cerebellar Purkinje cell has been the focus of numerous studies involving the analysis of development and information processing in the nervous system. Purkinje cells represent less than 0.1% of the total cell content of the cerebellum. To facilitate studies of molecules that are expressed in such a small proportion of neurons, we have established procedures for the purification of these cells. Transgenic mice were developed in which the expression of green fluorescent protein (GFP) was controlled by the L7 promoter. In adult cerebellum, GFP fluorescence was only detected in Purkinje cells, where it filled dendrites, soma and axons. GFP fluorescence was detected in Purkinje cells as early as embryonic day 17 and increased during development in vivo and in dissociated cerebellar culture. Mirroring endogenous L7 expression, high levels of GFP were observed in retinal rod bipolar cells. Lower levels of GFP were seen in olfactory periglomerular cells, neurons in the interpeduncular nucleus, and superior colliculus neurons. Cerebella from transgenic mice were dissociated by mild enzymatic treatment and Purkinje cells were isolated by fluorescence-activated cell sorting (FACS). By selecting optimal parameters, a fraction of viable Purkinje cells that was 94% pure was obtained. These results indicate that FACS is a powerful tool for isolating Purkinje cells from postnatal L7-GFP transgenic mice. GFP-positive neurons will also be useful in the real-time observation of dendritic morphogenesis and axonal outgrowth during development, or after neuronal activity in vitro.

Parkinson's disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta with accompanying evidence of increased oxidative damage, deficits in mitochondrial function and iron deposition. Recently, haem oxygenase-1 levels were reported to be elevated in PD brains. Because this enzyme is involved in the response to oxidative stress and is critical for cellular haem and iron homeostasis, it could play a role in the pathogenesis of PD. Therefore, we investigated the expression of haem oxygenase isoform 1 (HO-1) in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. MPTP triggered a relatively rapid and persistent increase in HO-1 mRNA exclusively in the mouse striatum. In situ hybridization and immunohistochemistry showed HO-1 to be localized to striatal astrocytes. The induction of HO-1 by MPTP was blocked by selegiline and GBR-12909, indicating the protoxin had to be metabolized by monoamine oxidase B and taken up by dopaminergic neurons to exert its action in astrocytes. MPTP did not alter the expression of other enzymes of haem synthesis or degradation nor were the levels of mRNA for haem or iron-binding proteins changed. Thus, expression of HO-1 was not part of a cellular program involving haem biosynthesis or homeostasis. In addition, heat shock proteins were not induced by MPTP. Thus, MPTP elicited a selective transcriptional response in striatal astrocytes. This response appears to be mediated by molecules released from affected dopaminergic nerve terminals in the striatum acting upon neighbouring astrocytes. This signalling pathway and its potential relevance to PD are discussed.

Changes in abundance and certain population parameters of American plaice Hippoglossoides platessoides in Northwest Atlantic Fisheries Organization (NAFO) subdivision 3Ps, south of Newfoundland, were examined over the period 1972–1994. During these years, there has been a substantial reduction in population size and in more recent years, there has been a redistribution of the remaining population to deeper, warmer water than previously observed. Despite the substantial changes in stock size and distribution, there is no clear evidence that growth patterns have changed nor has there been any truncation of the age structure of the population, both of which commonly occur if overexploitation takes place. On the other hand, age and size at 50% sexual maturity for both males and females declined over the same period. This stock has been managed through quota regulation since 1974; however, the quota rarely restricted the fishery. During the 1980s, when the greatest declines in stock abundance occurred...

It has been suggested that Purkinje cells (PC) play a role in organizing topographic relationships of several cerebellar afferent systems, including olivocerebellar fibers. This hypothesis is based on the observation that PC in the rat express biochemical heterogeneities during the presumptive period of olivocerebellar fiber ingrowth to the cerebellum. Previous studies designed to investigate the organization of murine olivocerebellar fibers during embryogenesis have suggested that interactions with PC may play a role in segregating olivocerebellar fibers after they enter the cerebellum. To determine whether PC heterogeneities are related to olivocerebellar fiber organization, transgenic mice carrying a beta-galactosidase (beta-gal) reporter gene linked to the promoter from the PC-specific gene L7/pcp-2 were used in neuroanatomical tracing experiments. Expression of the transgene mirrors endogenous L7/pcp-2 expression, which is upregulated earliest in parasagittal strips of the vermal cortex. Studies were conducted in vitro by using brainstem-cerebellar explants from embryonic day 17/18 (E17/18) and 18/19 mice. Applications of neuroanatomical tracer (horseradish peroxidase or neurobiotin) were made in either the caudal medial accessory olive (cMAO) or the rostral olive. These studies indicate that groups of olivocerebellar fibers and clusters of L7/lacZ+ and L7/lacZ-Purkinje cells respect common distribution boundaries during late embryogenesis. The strong correspondence between the distribution patterns generated by these two markers suggests that expression of L7/pcp-2 and the topographic organization of olivocerebellar (OC) fibers are not interdependent, but may be regulated by a common event or interaction, of a presently unknown nature, which occurs earlier during cerebellar development.

In the past several years a great deal of evidence has accumulated linking neuronal activation events to the regulation of gene expression. We have pursued an analysis of c-fos regulation in the nervous system to elucidate the molecular mechanisms involved in stimulus-transcription coupling. The c-fos gene can be viewed as an archetype of the set of cellular immediate-early genes encoding transcription factors. These genes are believed to function in coupling short-term signals elicited by extracellular to long-term changes in cellular phenotype by orchestrating alterations in target gene expression. Several animal seizure models have been used to demonstrate the activation of gene expression in specific populations of neurons. Using a transgenic mouse approach, based on a foslacZ fusion gene, we now demonstrate an association between c-fos expression and cell death in the nervous system. A delayed and protracted induction was observed following surgical lesion and in response to neurotoxin exposure. This system allows us to determine, for the first time, the DNA regulatory sequences that are responsible for the induction of gene expression in neurons in vivo. Furthermore, foslacZ transgenic mice provide a unique resource for identifying cell populations that respond to specific stimuli or that are susceptible to particular toxins. © 1995 John Wiley & Sons, Inc.

A small acidic polypeptide, termed thymosin beta 10, has been identified and is present in the nervous system of the rat by the ninth day of gestation. Thymosin beta 10 levels rise during the remaining days of life in utero, and then decline to nearly undetectable values between the second and fourth week post partum. The present study investigates the possible developmental signals and mechanisms that might regulate the expression of thymosin beta 10 during neuroembryogenesis. Many cell lines derived from tumors of the central nervous system express thymosin beta 10, as well as its homologue gene product, thymosin beta 4. Because some of these cell lines respond to exogenously applied agents by increasing their apparent state of differentiation, we have determined whether thymosin beta 10 levels are coordinately modulated. In several neuroblastomas, including the B103 and B104 lines, retinoic acid elicits a time- and dose-dependent increase in the content of thymosin beta 10, but not that of thymosin beta 4. The increase in thymosin beta 10 polypeptide is associated with a marked increase in the specific mRNA encoding this molecule. The mRNA for thymosin beta 4 is unaffected by retinoic acid. This is in contrast with the situation in vivo, where the expression of both genes decreases after birth. Other agents that influence the morphology of B104 cells, such as phorbol esters and dibutyryl cyclic AMP, have no influence on beta-thymosin levels. A range of steroids, which like retinoids act upon nuclear receptors, was also inactive. The stimulatory action of retinoic acid is detectable within 4 h, and thymosin beta 10 peptide levels continue to rise for at least 4 days. The influence of the isoprenoid is fully reversible and exhibits structural specificity. We believe that this culture system is mimicking the early rising phase of thymosin beta 10 levels in brain and that endogenous retinoids may be candidate physiological regulators of this gene.

HPLC analysis of guanidinium hydrochloride extracts of neonatal and adult rat brain revealed a polypeptide that is present in high concentration in the immature nervous system, but whose levels decline dramatically in the adult. This polypeptide has been isolated and its complete amino acid sequence determined by gas-phase Edman degradation following specific chemical and enzymatic cleavages. The molecule is identified as thymosin beta 10, a member of a multigene family that encodes a structurally conserved series of small acidic polypeptides of uncertain function. Thymosin beta 10 is present in the developing nervous system as early as embryonic day 9. Levels subsequently increase to peak values between embryonic day 15 and postpartum day 3, before falling to adult values (about a 20-fold reduction) by postpartum day 14. The elevated levels of thymosin beta 10 in fetal and neonatal brain correlate with high levels of thymosin beta 10 mRNA, whereas the low values of the polypeptide in the adult and juvenile are mirrored by an approximate 15-fold reduction in specific mRNA. In comparison, the levels of thymosin beta 4 polypeptide, a homologue of thymosin beta 10, only decline by about 20% during the same developmental period. However, the mRNA encoding thymosin beta 4 is elevated in fetal brain, and its levels decrease approximately four-fold to a stable value around the time of birth. The reason for this discrepancy between thymosin beta 4 protein and mRNA levels is unknown. Thymosin beta 10 can also be detected by HPLC in fetal liver, where levels are approximately 5% of those in brain. In liver, thymosin beta 10 also declines following birth. It is concluded that beta-thymosin expression (as measured by steady-state mRNA and polypeptide levels) is both up- and down-regulated during different phases of maturation of the mammalian nervous system.

Periodic induction of focal electrical seizure [afterdischarge (AD)] is an absolute prerequisite for the development of kindling, an animal model of complex partial epilepsy. Once established, it is a permanent condition. The mechanism(s) that translate ADs, which last tens of seconds, into life-long alterations in the CNS is unclear. Cellular immediate-early genes have been implicated in the conversion of short-term stimuli to long-term alterations in cellular phe-notypes by regulating target gene expression. We have investigated the contribution of one such early gene, c-fos, to this process. The relationship between ADs and expression of c-fos gene in the rat hippocampus, a key structure in kindling development, was studied by analysis of mRNA levels. The low constitutive expression of c-fos mRNA in the hippocampus was not altered by kindling. There was an “all-or-none” relationship between induction of c-fos and the duration of AD. The threshold for induction was approximately 30 s of AD. Above-threshold ADs induced c-fos in both naive and kindled animals to the same extent and with identical temporal profiles. Although the expression of c-fos is unchanged with kindling, c-fos may nonetheless contribute to many long-term changes of kindling, both adaptive and epileptogenic.

A novel device killed with steam most surface organisms on fresh meat without cooking the surface. Treatment for 25 milliseconds with 145°C thermally saturated steam in the absence of air, followed by vacuum cooling, produced a 4 log kill of an applied dose of 10 7 L. inocua on raw fresh chicken meat. Similar results were achieved on fresh beef and pork. Total cycle time

Career development is central to men's lives because occupation is closely welded to male identity. Specific issues discussed in this article include: sex role stereotypes and men's careers; the male equation of occupational success and high self esteem; the balancing of work, personal attachments and recreation; traditional and nontraditional careers; male-female employment combinations; developmental issues of mid-life and retirement; and high consumption (the result/motivator of occupational success?). Men will continue to need clearly conceptualized and highly skilled help with career development issues.

A set of mouse monoclonal antibodies has been generated against rabbit muscle camosine synthetase. The immunoreactivity of these antibodies has been characterized using an immunoassay that permits the separation and direct measurement of the synthetase activity on a second antibody bead complex. Four IgG monoclonal antibodies bind the carnosine synthetase activity from muscle of all mammals tested (mouse, rat, rabbit, cow, dog, and monkey) but not that from chicken muscle. This indicates the mammalian enzymes share epitopes that are absent from the avian enzyme. In addition, relative tissue levels of synthetase activity can be quantified with this immunoassay. Thus, high levels of carnosine synthetase activity are immunoprecipitated from the olfactory tissues of both rat and rabbit. Synthetase activity is generally lower in other tissues (muscle, brain, heart, liver, and gut). Nevertheless, the cross-reactivity of the synthetase from several tissues (olfactory mucosa, muscle, brain, gut, heart, and liver) of a single species indicates the enzyme protein contains similar epitopes in these tissues. Immunoaffinity purification of this low-abundance, unstable enzyme should now be possible for subsequent studies of structure and regulation.

Depolarization of neurons either in culture or in vivo results in the rapid, calcium-dependent induction of several, so-called, immediate-early genes; the prototypes being c-fos and c-jun. The proteins encoded by c-jun, c-fos, and several fos-related genes all participate in a complex that interacts with the AP-1 consensus DNA sequence, previously shown to be important for the "transcriptional activation" of certain genes. Thus it is proposed that neuronal stimulation, via elevated intracellular calcium, leads to the induction of a series of genes, some of which encode proteins involved in transcriptional regulation, that contribute to long-term adaptive and plastic responses. Surprisingly, the molecular composition of the brain AP-1 binding complex varies with time after stimulation. This is because some of the inducible Fos-related proteins accumulate with much slower kinetics than Fos itself and only appear in significant amounts when Fos has disappeared. Of some considerable interest is the result these compositional alterations have upon the transcriptional activity of the AP-1 complex. Given the foregoing findings we consider some of the possible implications this might have for aging and neurodegenerative disorders particularly with regard to alterations in cellular calcium homeostasis.

Induction of c-fos expression occurs following treatment of diverse cell types with agents that trigger mitogenesis, differentiation or membrane depolarization. We suggest that c-fos may be regarded as a marker for a set of rapidly induced genes (termed cellular immediate-early genes) whose function is to couple extracellular stimulation to long-term responses. In the brain, these genes may contribute to the adaptive alterations involved in neuronal plasticity.

A nonganglioside factor(s) present in Sigma types II and III mixed bovine brain ganglioside preparations synergises with suboptimal amounts of serum to induce proliferation specifically in nondividing B 103 neuroblastoma cultures. The active substance is nondialysable and soluble in water as well as in chloroform-methanol mixtures of 1:1-4:1 (vol/vol). It is completely insoluble in ether and acetone at room temperature. Biological activity survives heating to 70 degrees C in the presence of 0.1 M HCl for 1 h as well as boiling at neutral pH. Loss of activity occurs on heating to 70 degrees C for 1 h with 1 M HCl or 1 M NaOH. The activity is insensitive to digestion with neuraminidase, trypsin, pronase, and phospholipases A2 and C. The factor cochromatographs with gangliosides on Dowex AG 50W and Sephadex G100 and is partially recovered with GM1 on DEAE-Sepharose, but may be isolated in a ganglioside-free fraction by sequential chromatography on Sephadex LH20 and silicic acid columns. The substance(s) has the properties of a water-soluble proteolipid protein, the amino acid composition being reported. It is not immunologically cross-reactive with antibodies to GM1 ganglioside or the major proteolipid protein of myelin.

The use of imagery in career counseling and life planning continues to increase just as the use of imagery in counseling and psychotherapy increases. The rationale for using imagery in career counseling and life planning is the same as in counseling and psychotherapy: It allows the client and counselor to bring into awareness unconscious material that is already influencing choices and makes this material usable in future planning. Imagey also allows the client to try on alternative roles. Several studies are cited that indicate that the imagery components of daydreams and fantasies are useful tools for career exploration and life planning when they are accessible to clients and properly processed. Relevant counseling applications and research studies are cited. Some recent tools for eliciting and processing these daydreams and fantasies are presented, and limits as well as benefits of techniques are discussed.

A new model has been established for the perfused in situ rat lung. During perfusion with glucose at 4 mmol/l, physiological concentrations of insulin (50 mu/l) caused an initial 1.5 fold increase in lactate production; the increase being sustained at 30% for the 4 h of perfusion. The observation that the lung responds to physiological concentrations of insulin could have important implications with respect to whole body carbohydrate metabolism.

This report describes results of a survey undertaken at the Western Regional Research Laboratory in cooperation with the University of California at Riverside. Cabbage, carrots, corn, lettuce, strawberries, and tomatoes harvested from plants grown in (1) clean air (carbon-filtered air); (2) clean air fumigated with a low ozone level; and (3) clean air fumigated with a high ozone level were studied. The two ozone levels used (approximately 200 and 350 ppB O/sub 3/) represented levels commonly observed in the air in Southern California in summer and fall. Items determined quantitatively included five vitamins, solids, nitrogen, fiber, ash carbohydrate, and up to nine metals. The metals Ca, Cu, Fe, Pb, Mn, K, Rb, Sr, and Zn were determined using X-ray fluorescence spectroscopy. With the metals significant differences occurred with different plants. The nonmetal nutrients showed that ozone had some effect on the crops. Carotene, vitamin C, thiamine, and riboflavin were studied. This survey indicated that ozone did not have a major and generally deleterious impact on crop composition. It indicated some areas where ozone influenced crop composition. 14 references, 5 tables.

SUMMARY Various additives were examined as possible aids in making bread from sorghum and barley flours. Good rise was achieved with doughs containing 45% solids. Several gums, especially 4000 centipoise methylcellulose, increased gas retention in sorghum bread and improved texture of both sorghum and barley breads. Several starches improved texture and loaf volume of sorghum bread. Glyceryl monostearate (GMS) improved the texture of sorghum bread but caused it to crumble badly. GMS improved softness in barley bread. Shortenings also softened sorghum and barley breads. The effect of several processing variables on sorghum bread was studied along with 2 methods of imparting a sour fermented flavor to sorghum bread.

SUMMARY An oil with a characteristic strawberry aroma was obtained from the condensate from the jam-making process. Programmed-temperature capillary gasliquid chromatography (PTCGLC) showed the oil to be a very complex material of more than 150 components. A fast-scan mass spectrometer was used to analyze the material as it was eluted from the PTCGLC equipment, and some of the more volatile compounds identified are reported.

SUMMARY A combined feedforward-feedback system is proposed to reduce fouling in a vertical long-tube tomato evaporator. Development of the appropriate heat and mass transfer equations under conditions of changing feed composition and constant evaporator temperature indicated that perfect control can theoretically be maintained by feedforward control of flow rate. Derivative and proportional control are required. Fouling in the evaporator tubes and uncertainties of process dynamics necessitate the addition of integral and proportional feedback control to the feedforward system. The proposed evaporator-control system was simulated on an analog computer. The results proved that excellent control of the product can be maintained in single-pass operation with feedforward-feedback control.

SUMMARY A feedforward-feedback control system has been successfully combined with a vertical long-tube tomato evaporator. The product solids concentration from this pilot plant unit was stabilized to ± 0.1% solids despite 10% feed concentration variations. These results were obtained by varying the feed rate instead of the evaporator temperature. Signals from two in-line refractometers continuously measuring solids in feed and product were fed to an analog computer. The computer output was then used to vary the feed rate through an electromechanical linkage.

SUMMARY —Changes in the free amino acid fraction of foam-mat dried tomato powder during storage were studied using gas-liquid chromatography. An increase of individual amino acids was found to be independent of storage temperature and possibly caused by proteolytic reaction. A simultaneous decrease in certain amino acids was accelerated at higher storage temperatures. This loss of amino acids may be involved in non-enzymatic browning reactions.