Regional changes in forebrain activation during the early and late phase of formalin nociception: analysis using cerebral blood flow in the rat

This is the first neural imaging study to use regional cerebral blood flow (rCBF) in an animal model to identify the patterns of forebrain nociceptive processing that occur during the early and late phase of the formalin test. We measured normalized rCBF increases by an autoradiographic method using the radiotracer [99mTc]exametazime. Noxious formalin consistently produced detectable, well-localized and typically bilateral increases in rCBF within multiple forebrain structures, as well as the interpeduncular nucleus (Activation Index, AI = 66) and the midbrain periaqueductal gray (AI = 20). Structures showing pain-induced changes in rCBF included several forebrain regions considered part of the limbic system. The hindlimb region of somatosensory cortex was significantly activated (AI = 31), and blood flow increases in VPL (AI = 8.7) and the medial thalamus (AI = 9.0) exhibited a tendency to be greater in the late phase as compared to the early phase of the formalin test. The spatial pattern and intensity of activation varied as a function of the time following the noxious formalin stimulus. The results highlight the important role of the limbic forebrain in the neural mechanisms of prolonged persistent pain and provide evidence for a forebrain network for pain.     

Fine structural changes of the porcine uterine tube epithelium during early and late pregnancy

Ultrastructural changes in the uterine tube (oviduct) of pregnant gilts have been investigated with special reference to the ciliated, secretory, and stromal cells. Tissue from the uterine tube ampulla and infundibulum was taken from 18 gilts at different stages of gestation (days 31, 36, 101, 102, 107, 110, and 112). Cilia were present throughout pregnancy, and deciliation was not apparent at any stage of gestation. The low epithelium of the uterine tube appeared similar to that of the luteal phase of the estrous cycle when corpora lutea were full grown. Prominent features at end of the gestation were numerous fibrous granules and basal bodies, indicating active formation of ciliary precursor organelles. Fibrogranular aggregates were also present in association with the basal bodies. In addition, numerous polyribosomes, mitochondria, and microtubules were encountered in the cytoplasm of ciliated cells at end of the gestation. The appearance of electron-opaque, fibrous granules during late pregnancy probably could be correlated with increasing endogenous levels of plasma estrogen. Intimate morphologic association between fibrous granules and basal bodies indicate that fibrous granules might provide precursor material for the development of cilia or rootlets. Characteristics ultrastructural changes observed in secretory cells during the estrous cycle were not discernible in secretory cells during pregnancy. The secretory cells appeared similar to those of the luteal phase of the estrous cycle. The apocrine secretory cells contained prominent, apical, cytoplasmic projections; pinching-off process of these protrusions was frequently observed during early and term gestation. Extruded nuclei along with other cytoplasmic organelles were also present, lying free in the tubal lumen. The endoplasmic reticulum was predominantly tubular in form. Synthesis, storage, and release of secretory granules were not apparent at early or late pregnancy. It is suggested that progesterone might have an inhibitory effect on the synthesis, storage, and release of secretory granules. Ultrastructural changes in stromal cells were not apparent at any stage of gestation. The stromal cells appeared similar to that of the luteal phase of the estrous cycle.     

An evolutionarily conserved region in the second intron of the human nestin gene directs gene expression to CNS progenitor cells and to early neural crest cells

Central nervous system (CNS) progenitor cells transiently proliferate in the embryonic neural tube and give rise to neurons and glial cells. A characteristic feature of the CNS progenitor cells is expression of the intermediate filament nestin and it was previously shown that the rat nestin second intron functions as an enhancer, directing gene expression to CNS progenitor cells. In this report we characterize the nestin enhancer in further detail. Cloning and sequence analysis of the rat and human nestin second introns revealed local domains of high sequence similarity in the 3' portion of the introns. Transgenic mice were generated with the most conserved 714 bp in the 3' portion of the intron, or with the complete, 1852 bp, human second intron, coupled to the reporter gene lacZ. The two constructs gave a very similar nestin-like expression pattern, indicating that the important control elements reside in the 714 bp element. Expression was observed starting in embryonic day (E)7.5 neural plate, and at E10.5 CNS progenitor cells throughout the neural tube expressed lacZ. At E12.5, lacZ expression was more restricted and confined to proliferating regions in the neural tube. An interesting difference, compared to the rat nestin second intron, was that the human intron at E10.5 mediated lacZ expression also in early migrating neural crest cells, which is a site of endogenous nestin expression. In conclusion, these data show that a relatively short, evolutionarily conserved region is sufficient to control gene expression in CNS progenitor cells, but that the same region differs between rodents and primates in its capacity to control expression in neural crest cells.     

Evidence for a phase transition in the early development of prehension

The genotoxicity of twenty one clinically used (or discontinued) antihistamines is reviewed. New results are also presented from an evaluation of selected antihistamines in the V79 in vitro micronucleus assay. For two antihistamines, no genotoxicity data is available. Of the remaining nineteen, nine have been reported as positive and one equivocal in at least one genotoxicity assay despite the fact that none possess structural alerts for genotoxicity. Ethidium displacement and bleomycin amplification studies in V79 cells indicate that nine of these ten antihistamines are capable of intercalative DNA binding. Further, nine of the ten positive compounds, but none of the tested compounds which also intercalate but are reported to be negative in gene-tox assays (e.g. triprolidine, chlorcyclizine, clemastine), possess a dimethylamino substituent suggesting the requirement for this cationic function in the genotoxicity. It is proposed that the apparent genotoxicity of antihistamines and possibly many other pharmaceuticals derives from a hitherto unappreciated propensity of these drugs for stabilized intercalative DNA binding. It is further proposed that the bleomycin amplification assay may provide a widely applicable means for assessing functional intercalative drug/DNA interaction in intact mammalian cells.     

Differential regulation of early phase and late phase responses in human neutrophils by cAMP

The elevation of intracellular levels of cyclic AMP by forskolin stimulation of adenylate cyclase regulates early and late phase neutrophil responses differentially. Early phase neutrophil responses as measured by shape change in response to chemotactic factors, transmigration across a polycarbonate membrane and priming were unaffected by forskolin-induced elevation of intracellular cAMP. Late phase neutrophil responses such as release of superoxide anions, activation of phospholipase A2 and platelet activating factor (PAF) synthesis were inhibited by increasing intracellular cAMP through the addition of 10 microM forskolin for 10 min prior to stimulation. N-Formyl-methionyl-leucyl-phenylalanine-stimulated arachidonic acid release fell from 9.3% (untreated cells) to 4.6% in forskolin-treated cells. PAF generation was also inhibited from 430 pg/10(6) cells in untreated cells to background levels in forskolin-treated cells (110 pg/10(6) cells). Also, the reduction of cytochrome c by superoxide anions fell from 4.2 nmol/10(6) cells in the absence of forskolin to 2.0 nmol/10(6) cells following forskolin treatment. These results indicate that in neutrophils the elevation of cAMP acts differentially on cellular responses, not affecting early activation events, but markedly inhibiting late events such as the release of inflammatory mediators.     

Dynamics of changes in anomalous human cells during prolonged cultivation in the stationary phase. Trisomy 7 cells

Ontogenetic changes of normal diploid cells and cells with chromosomal aberration (strain LHC-162; 47, XY, +7) were studied in prolonged cultivation in stationary phase. Cultivated cells were human found to possess their specific type of ontogenetic changes for each culture; the dynamics of these changes depended on the density of cellular population. Two morphologically different cellular subpopulations differing by the degree of cell nucleus heterochromatinization (normal strains) and three different cellular subpopulations in the aberrant strain (47, XY, +7) were discovered in studying the cell nucleus.     

Interferons impair early transgene expression by adenovirus-mediated gene transfer in muscle cells

Sarcosine reductase is the only reductase system present in Tissierella creatinophila when grown on creatinine plus formate. The acetyl-phosphate-forming component protein C was purified to homogeneity. SDS-PAGE of the purified protein revealed two protein bands with apparent mol. masses of 62 and 50 kDa. The N-terminal amino acid sequence of the two subunits was determined. Antibodies raised against each of the subunits of protein C from Eubacterium acidaminophilum cross-reacted with the corresponding protein present in T. creatinophila, Clostridium litorale and Clostridium sporogenes. The arsenate-dependent hydrolysis of acetyl phosphate catalyzed by protein C was partly inhibited by antibodies directed against the large subunit. Antibodies raised against the small subunit were twice as effective, which indicates that this subunit is the primary site of acetyl transfer from acetyl phosphate. The protein A component of the sarcosine reductase of T. creatinophila was purified to homogeneity by cochromatography with thioredoxin reductase on DEAE-Sephacel, hydroxylapatite, Q-Sepharose, and Sephacryl 100-HR. Protein A had an apparent mol. mass of 21 kDa. Its N-terminal amino acid sequence showed high similarities to that of other proteins A. Initial steps for the purification and preliminary characterization of the sarcosine-specific, substrate-binding protein Bsarcosine component of T. creatinophila indicated the involvement of a 50-kDa protein.     

Decreased IGF-I gene expression during the apoptosis of Purkinje cells in pcd mice

The purpose of this study was to develop concepts that facilitate our understanding of why family caregivers of demented elderly persons can continue caregiving despite various difficulties of care. Twenty-six Japanese daughter or daughter-in-law caregivers of elderly parents with dementia who lived at home or in long-term care facilities were recruited through various senior service organizations in Japan. The caregivers underwent unstructured interviews, and the interview data were analyzed using the constant comparative method. Three categories emerged as reasons for care continuation: value of care, maintainers of value, and reinforcers of care continuation. Value of care came from societal norms and attachment, and was the basis of caregivers' motivation to continue care. Several maintainers of value and reinforcers of care continuation also emerged from the analysis. The contents and some longitudinal changes in these categories were explained. The findings highlight the need to assess these categories separately in order to develop appropriate interventions and they have implications for future research and policy development.     

Control of gene expression in Xenopus early development

Risk adjustment is intended to minimize selection of patients or enrollees in health plans. Current efforts generally are recognized as inadequate, but improvement is difficult. The greatest short-term gain will come from introducing diagnostic information, though outpatient diagnosis data are unreliable. Initial efforts may use inpatient data, but this creates incentives to hospitalize people. Even exploiting diagnosis information leaves substantial imperfections. Partial capitation, common in behavioral health, reduces incentives to select patients and stent on services, but current policy resists it, perhaps because policymakers misinterpret the lesson of the Prospective Payment System. Theoretically, not paying plans more for providing additional services is optimal only if consumers are well informed.     

Human obese gene expression: alternative splicing of mRNA and relation to adipose tissue localization

The aim of this work was to obtain further information about the origin of fluoride profiles in cementum. Fluoride was administered to rats at varying doses (0.50, 100 ppm F in drinking water) and for different durations (4, 13 and 25 weeks). Fluoride distribution across the full thickness of molar cementum in rats was measured by means of an abrasive micro-sampling technique. The average fluoride concentrations in cementum increased significantly with increasing dose and duration of fluoride administration. The relative reduction of the average fluoride concentrations after cessation of fluoride administration was 94.2-36.5% at 50 ppm F and 62.2-49.2% at 100 ppm F in the outer layers (1-60 microns) and 91.5-24.1% at 50 ppm F and 74.1-7.6% at 100 ppm F in the middle (61-120 microns) layers of the cementum, respectively. The reduction rates were more closely related to the time intervals following cessation rather than fluoride concentrations in drinking water or specificity within the cementum. Two factors which may influence this are new cementum formation after withdrawal of fluoride and some fluoride release from cementum surfaces when the fluoride supply stopped. It was concluded that the cessation of fluoride administration reduced the fluoride concentration on the outer layers of cementum differing from bone where reduction occurs across the entire thickness.     

Novel mutations in the TIGR gene in early and late onset open angle glaucoma

Chronic experiments on four dogs were performed to study the effects of bilateral microinjection of the choline receptor agonist carbacholine and the blocking agent scopolamine into the dorsolateral part of the head of the caudate nucleus on the performance of an operant defensive reflex involving maintenance of a specified amount of flexion and on the differentiation of meaningful signals. Bilateral microinjection of carbacholine (0.1-0.4 micrograms in 1.5 microliters of solvent) reduced the phasic component and amplified the tonic component of the operant responses, inhibited intersignal leg lifts, normalized posture, and calmed the animals, and also led to a sharp improvement in the differentiation of meaningful signals. These changes were expressed as increases of three-fold or more in the latent period of movement initiation when the differentiation signal was used, as compared with the baseline latent period before the injections. Microinjection of the choline receptor blocker scopolamine into the striatum had the opposite effects. Unilateral microinjection of these substances produced changes mainly only on the day of dosage and had no effect on subsequent behavior, while bilateral microinjection altered the established motor behavior for a longer period of time. This affected both the motor and the sensory components of the operant response.