Descending neural projections to the spinal cord in the channel catfish, Ictalurus punctatus

Retrograde transport of horseradish peroxidase was used to determine the descending projections to the spinal cord in an otophysan fish, the channel catfish, Ictalurus punctatus. The majority of cells projecting to the spinal cord are located in the reticular formation, which is organized into rhombomeric segments. Vestibulospinal neurons are located in the descending, magnocellular, and tangential octaval nuclei, as well as in the medial octavolateralis nucleus of the lateral line system. Cells in the facial lobe project to the spinal cord. Additionally, axons of cells of the trigeminal system and the nucleus of the lateral lemniscus project caudally into the spinal cord. In the midbrain, descending spinal projections arise from cells of the medial longitudinal fasciculus and the red nucleus. More rostrally, cells of the ventrolateral thalamus, dorsal periventricular hypothalamus, central pretectal and magnocellular preoptic nuclei also project to the cord. The results of this study indicate that there are a number of homologies in the descending systems of bony fishes and other vertebrate taxa, including tetrapods. We also provide further evidence that a red nucleus is present in the brains of bony fishes and is therefore a primitive vertebrate character antedating the evolution of tetrapods.     

Immunotoxicity in channel catfish, Ictalurus punctatus, following acute exposure to tributyltin

Tributyltin (TBT) is a trialkylated organotin formulated for use primarily as a biocide for aquatic and agricultural industries. Although macrophages isolated from toadfish (Opsanus tau) are sensitive to exposure, very little is known about the effects of TBT on fish humoral immunity and non-specific cytotoxic cell (NCC) functions. To evaluate the effects of TBT on these parameters, channel catfish (Ictalurus punctatus) were given a single intraperitoneal injection of corn oil with 0 (vehicle control) or with 0.01, 0.1, and 1.0 mgTBT/kg as TBTCl. Three and seven days later, NCC activity and phagocyte oxidative burst were evaluated and related to allometric indices and hematology. The humoral immune response to Edwardsiella ictaluri was evaluated fourteen days after treatment. Peripheral blood neutrophilia and specific antibody secreting cell (SASC) numbers were the most sensitive parameters and were affected in all three TBT treatment groups. Allometric indices, peripheral blood lymphocyte and monocyte percentages, NCC activity, and phagocyte oxidative burst were less sensitive and were affected only at the highest dose of TBT.     

Identification and characterization of a heat shock protein 70 family member in channel catfish (Ictalurus punctatus)

The primary objective of this study was to evaluate the electrophysiologic effects of large-dose propofol, used as the sole anesthetic in patients with epilepsy. Nine patients with medically intractable complex partial epilepsy undergoing a three-stage approach to the surgical management of epilepsy were recruited. State I involved placement of the intracranial electrode array, while Stage II consisted of extraoperative localization of the seizure focus. The patients were studied during induction of anesthesia for Stage III (removal of electrodes and resection of seizure focus). Unpremedicated patients were induced with a propofol infusion (0.5 mg.kg-1.min-1) until one of the following occurred: 1) electrical seizure activity, 2) burst suppression, or 3) total dose of 10 mg/mg. Electrocorticography (ECoG) was recorded continuously during this period. Two patients were excluded from the study after experiencing delayed awakening after the Stage I procedure. Both had received propofol along with other anesthetics. No ECoG evidence of seizure activity was detected in the seven patients completing the study. Burst suppression was attained in six patients using a mean dose of 5.7 mg/kg +/- 2.6. We conclude that large dose propofol alone does not trigger electrical epileptiform activity on the ECoG of seizure patients.     

Electrophysiological responses of single olfactory bulb neurons to binary mixtures of amino acids in the channel catfish, Ictalurus punctatus

1. For the first time in any vertebrate, responses of single olfactory bulb neurons to odorant mixtures were studied quantitatively in the channel catfish, Ictalurus punctatus. 2. Extracellular electrophysiological responses of 61 single olfactory bulb neurons from 36 channel catfish to binary mixtures of amino acids and to their components were recorded simultaneously with the electro-olfactogram (EOG). Tested were a total of 297 mixture trials consisting of 18 different stimulus pairs formed from 8 amino acids. 3. For 42% (126 of the 297) of the tests, no significant change (N) from spontaneous activity occurred. Responses to the remaining 171 tests of binary mixtures were excitatory (E; 29%) or suppressive (S; 29%). No response type was associated with any specific mixture across the neurons sampled. 4. Mixture interactions that changed response types (E or S) from those observed to the individual components were rare, because 89% of the responses of single olfactory bulb neurons to the tested binary mixtures were classified similarly as the responses to at least one of the components. 5. Responses of single olfactory bulb neurons were generally predictable for binary mixtures whose component responses were classified as both E, both S, and both N. For binary mixtures whose component responses were classified differently (e.g., one component evoked excitatory responses and the other evoked suppressive responses), the predictability of the response was dependent on the specific mixture type.     

Intracellular signaling events in superoxide generation and adhesion of channel catfish, Ictalurus punctatus rafinesque, neutrophils to the extracellular matrix protein fibrinogen

Cytosine deaminase is an enzyme which has been investigated for cancer chemotherapy as a result of its ability to convert the relatively nontoxic prodrug 5-fluorocytosine into the anticancer drug 5-fluorouracil. To facilitate investigations of the utility of cytosine deaminase for cancer chemotherapy, we have cloned and expressed the enzyme from Saccharomyces cerevisiae. The DNA sequence translates into a protein of 158 amino acids in length, with a predicted molecular weight of 17,563 kilodaltons. Alignment of the cytosine deaminase protein sequence from yeast with a variety of proteins defines a novel sequence motif of cytosine or cytidine binding enzymes. Recombinant expression cassettes encoding cytosine deaminase were transfected into monkey kidney COS cells, which lack endogenous cytosine deaminase, to test for production of a functional protein. Cell extracts from these transfectants contained detectable levels of enzyme activity capable of converting 5-fluorocytosine to 5-fluorouracil. Cytosine deaminase was expressed in yeast from a cDNA cassette under the control of an inducible promoter, increasing expression 250- to 300-fold relative to wild-type strains. A purification protocol has been developed which permits recovery of 60% of cytosine deaminase in active form from induced cell lysates after two purification steps. This protocol will be useful for isolating large quantities of pure enzyme which are required for the preclinical evaluation of monoclonal antibody-cytosine deaminase conjugates in combination with 5-fluorocytosine.     

Dietary modulation of phase 1 and phase 2 activities with benzo(a)pyrene and related compounds in the intestine but not the liver of the channel catfish, Ictalurus punctatus

EPA, DHA, C15SCH2COOH (n-3), C15SCH2COOH (n-6) and C18SCH2COOH (n-3) are extensively incorporated into phospholipids and triacylglycerol in rat hepatocytes after 24 h incubation with 80 microM fatty acid/derivative. Only traces of polyunsaturated 3-oxa fatty acids (C15OCH2COOH, C18OCH2COOH) were incorporated. C15-S-butyric acid (n-3) is a stronger inhibitor of delta6-desaturase in rat liver-microsomes than C15SCH2COOH (n-3), C15-S-propionic acid (n-3), EPA and DHA. It inhibits delta5-desaturase in a similar manner to EPA and DHA. Arachidonic acid and C15SCH2COOH, (n-6) are better substrates for PGH-synthase than EPA and C15SCH2COOH, (n-3), showing the inhibitory effect of the n-3 bond. The n-3 polyunsaturated fatty acids, including the sulfur-substituted fatty acid derivatives, are poor substrates for PGH-synthase. However, they inactivate the PGH-synthase activity at least as efficiently as arachidonic acid. C15SCH2COOH (n-3), C15S(CH2)2COOH (n-3) and C18SCH2COOH (n-3) induce peroxisomal beta-oxidation more than EPA and DHA.     

Antimicrobial activity of the new carbapenem biapenem compared to imipenem, meropenem and other broad-spectrum beta-lactam drugs

The in vitro activity of biapenem was compared to that of imipenem, meropenem and other broad-spectrum beta-lactams. A total of 716 isolates from recent cases of clinical septicemia and an additional 137 stock strains possessing known beta-lactamases or other well-characterized resistance mechanisms were tested. The minimal concentrations inhibiting 90% of strains (MIC90) of Enterobacteriaceae species were for biapenem 0.03 to 1 mg/l and for imipenem 0.25 to 2 mg/l. No member of the Enterobacteriaceae was found to be resistant to biapenem. Biapenem and meropenem were the most active drugs against Pseudomonas aeruginosa, with an MIC90 of 1 mg/l. Biapenem was more active than ceftazidime against most gram-negative and gram-positive bacteria tested. Biapenem was as potent as imipenem against anaerobic bacteria (including Bacteroides fragilis), with an MIC90 of 0.25 mg/l. High MICs of biapenem were demonstrated for Xanthomonas maltophilia, oxacillin-resistant Staphylococcus spp. and Enterococcus spp. These species have demonstrated resistance to other carbapenems and to most of the newer cephalosporins. The results of this study, coupled with previously documented favorable qualities of biapenem, endorse further investigation of this broad-spectrum antibacterial agent for clinical use.     

Antimicrobial mixtures used by tissue banks for harvested skin: comparative in vitro activity

The activities of antimicrobial combinations from three geographically diverse skin/tissue banks used in the processing of skin/ tissue were compared using bacteria and yeast isolated from burn patients. All formulations showed 90% or more effectiveness against bacteria generally susceptible to antibiotics but were less effective (60-80%) when tested against bacteria resistant to specific antimicrobials. Anti-yeast activity was present when an appropriate antifungal agent was included in the combination. All formulations were stable for at least six weeks. Results of this study raise certain questions about the use of these antimicrobial combinations in contemporary skin/tissue banking and point the way toward areas for future study.     

Immunochemical characterization of hepatic cytochrome P450 isozymes in the channel catfish: assessment of sexual, developmental and treatment-related effects

The profiles of immunoreactive proteins recognized by antibodies raised against purified trout P-450 isoforms (CYP1A1, CYP2M1 and CYP2K1) were examined in channel catfish liver by Western blot analysis. Gender differences in basal expression of these isoforms, as well as responses to known inducers of mammalian isoforms (ethanol, beta-naphthoflavone and clofibric acid) and early life stage (3 and 6 months) profiles are described. Two similar protein bands were detected by Western blotting in mature untreated catfish with CYP2K1 and CYP2M1 antibodies. A third band is detected by anti-2K1 in fish treated with beta-naphthoflavone; this band was verified as CYP1A, with about twice the level of expression in males versus females. No difference between sexes was seen in the expression of the 51-kDa CYP2-reactive bands; however, a significant difference (female > male) was seen in the lower molecular weight CYP2 band (47-kDa). Ethanol treatment caused a dose-dependent decrease in the 47-kDa CYP2-reactive isoforms but no change in the 51-kDa band. Clofibric acid treatment caused an increase in both the 51-kDa CYP2 protein as well as in liver somatic index. Age-dependent changes in isoform expression were also detected in CYP2-reactive forms, with a novel protein (53-kDa) detected in 3-month-old fish. The results from this study provide insight into the regulation of constitutive catfish CYP isoforms and prepares a foundation for further examination of the biotransformation capabilities of an important aquatic species.     

Channel catfish virus: comparative replication and sensitivity of cell lines from channel catfish ovary and the brown bullhead

A cell line derived from channel catfish ovary tissue was compared with the brown bullhead (BB) cell line for their respective abilities to replicate and detect channel catfish virus (CCV). The channel catfish ovary cell line (CCO) produced cytopathic effects (CPE) more rapidly and detected CCV at higher dilutions than did the BB cell line. Production of CCV was more rapid in CCO cells than in BB cells, but the peak titers of the two lines were not significantly different. The CCO cell line was shown to be the more sensitive cell line for CCV research and diagnostics.     

Preparation, characterization and in vitro antimicrobial activity of ampicillin-loaded polyethylcyanoacrylate nanoparticles

In this paper, the experimental conditions for preparing ampicillin-loaded polyethylcyanoacrylate (PECA) nanoparticles are described. The effects of drug concentration and surfactant type in the polymerization medium on the particle size distribution and loading capacity were studied. The results of these studies show that only the type of surfactant has an impact on the nanoparticle dimensions. The release rate of ampicillin from PECA nanoparticles at pH 7.4 (extracellular value pH) performed either with and without esterases, show that the drug release is considerably increased in the presence of these exzymes. The results of drug release study at pH 1.1 (simulated gastric juice) are very interesting. This study has evidenced that the 70% of ampicillin is released quickly, while the remaining fraction is firmly incorporated in nanoparticles. The released ampicillin is quickly degraded in acid medium while the entrapped fraction is protected from acid degradation and afterwards, when nanoparticles reach the small intestine, can be readily released in the presence of esterases. This result could be exploited for the oral administration of the ampicillin-PECA system. Finally, studies of antimicrobial activity of prepared systems evidenced that ampicillin-loaded PECA nanoparticles exhibit an activity equal or higher than the free drug.     

Voltage-dependent anion channel proteins in synaptosomes of the torpedo electric organ: immunolocalization, purification, and characterization

Secretory granules are the cellular organelles mediating storage and regulated secretion of proteins. Their biogenesis involves sorting of secretory protein cargo and membrane constituents, which takes place at two distinct levels, the trans-Golgi network and the immature secretory granule. At both levels, sorting is accomplished by cargo aggregation and cargo-membrane recognition. Given not only the aggregative properties of the regulated secretory proteins but also the ability of lipids to form distinct membrane microdomains, self-organization of both lumenal and membrane constituents is proposed to play a crucial role in secretory granule biogenesis.     

Antimicrobial activity and spectrum investigation of eight broad-spectrum beta-lactam drugs: a 1997 surveillance trial in 102 medical centers in the United States. Cefepime Study Group

Anti-inflammatory studies were conducted on rats or mice using a crude hydroalcoholic extract of the aerial parts of Turnera ulmifolia and it's partitioned fractions, i.e. the aqueous, ethyl acetate and ethanolic fractions. The hydroalcoholic extract and it's fractions (aqueous and ethanolic) inhibited carrageenan-induced edema. However, only the ethanolic fraction was used in the other experiments due to it's yield. The extract also inhibited the cotton pellet granuloma and the increase of vascular permeability induced by histamine, 5-hydroxytryptamine and prostaglandin E2, but not that produced by bradykinin. The extract or the fraction did not present analgesic activity in the writhing test using acetic acid and did not reduce croton oil-induced ear edema in mice. When the ethanolic fraction and LPS were administered i.p. to Balb/C mice 72 h before blood or peritoneal fluid collection, no changes were observed in the white or total blood cell counts in the peripheral blood. On the other hand, changes were observed in both total and differential cell counts in the peritoneal exudate since all doses of the fraction reduced the number of total leukocytes (mainly lymphocytes) obtained from the peritoneal exudate. In contrast to nonsteroidal anti-inflammatory drugs, the administration of the hydroalcoholic extract or the ethanolic fraction alone did not potentiate gastric mucosal lesions induced by aspirin. The extract and the fraction inhibited the appearance of gastric lesions induced by indomethacin, ethanol and pylorus ligature, but not those induced by stress. As also observed with carbenoxolone, the ethanolic fraction increased the wall mucus in hypothermical-restraint stress-induced gastric lesions. The anti-ulcerogenic effect of the extract and of the ethanolic fraction may be related to an increase of mucosal defensive factors, such as prostaglandin and mucus. The anti-inflammatory actions of the extract and the fraction may be due to an inhibitory effect on histamine and cyclooxygenase II, but not on cyclooxygenase I, because the extract and it's fraction present both anti-inflammatory and anti-ulcerogenic effects. The major substances present in the ethanolic fraction are flavonoids which will be isolated and identified.     

Furazolidone disposition after intravascular and oral dosing in the channel catfish

1. The pharmacokinetics, tissue distribution and excretion of the nitrofuran drug furazolidone have been examined in the channel catfish. [14C]Furazolidone was administered by intravascular or oral routes in a single dosage of 1 mg/kg body weight. 2. A two-compartment pharmacokinetic model best described parent furazolidone concentrations in the plasma after intravascular dosing. Elimination of parent compound was extremely rapid, with a terminal half-life of 0.27h and total body clearance of 1901 ml/h/kg. 3. After oral dosing, furazolidone concentrations in the plasma were highest at 1 h and were below the limit of determination (< 20 ng/ml) at 5 h. The oral bioavailability of parent furazolidone administered in solution was 58%, compared with 28% in a feed mixture. 4. Concentrations of furazolidone and its metabolites were highest in the excretory tissues and lowest in the muscle after oral dosing. Parent furazolidone comprised 10% of the total 14C in the muscle at 8 h and was not detectable (< 1 ng/g) at 24 h; total 14C concentrations declined from 274 to 59 ng furazolidone equiv./g between 8 and 168 h. Non-extractable (bound) residues comprised 18% of total 14C in muscle at 8 h and 33% at 168h. 5. Renal excretion was the primary route of elimination of 14C residues and accounted for nearly 55% of the oral dose.     

Axonal projection patterns of neurons in the secondary gustatory nucleus of channel catfish

The second gustatory nucleus of teleost fishes receives ascending fibers from the primary gustatory center in the medulla and sends efferent fibers to several nuclei in the inferior lobe of the diencephalon. Similar to the corresponding parabrachial nucleus in birds and mammals, the secondary gustatory nucleus of catfish consists of several cytoarchitectonically distinct subnuclei which receive input from different portions of the primary gustatory nuclei. However, it is unclear how the subnuclear organization relates to the processing of gustatory information in the hindbrain and the subsequent transmission of that information to the forebrain. To determine whether cells within different subnuclei of the secondary gustatory nucleus of channel catfish project to different diencephalic targets, single cells were intracellularly labeled with biocytin. Three subnuclei have been identified in the secondary gustatory nucleus: a medial subnucleus spanning most of the rostrocaudal extent of the nucleus, a central subnucleus and a dorsal subnucleus, the latter two located in the rostrolateral portion of the complex. Cells throughout the secondary gustatory nucleus typically possessed similar collateral projections to several nuclei in the inferior lobe, although four of the six cells filled in the medial subnucleus projected only to nucleus centralis. The only apparent subnucleus-specific projection pattern involved cells at the rostral edge of the secondary gustatory nucleus and in the secondary visceral nucleus. Axons of these cells terminated only in restricted portions of nucleus lobobulbaris. These results suggest that efferents from different subnuclei of the secondary gustatory nucleus of catfish, like those of the parabrachial nucleus of birds and mammals, do not possess simple, topographical projections to target nuclei in the diencephalon.     

Accumulation and elimination of dieldrin in muscle tissue of channel catfish

Dieldrin accumulation and elimination in muscle tissue of channel catfish (Ictalurus punctatus) from water and food was determined in the laboratory. Twenty-eight-day exposure of fish 150-225 mm and 350-400 mm long to 75 parts per trillion (ng/liter) dieldrin resulted in the larger catfish consistently accumulating more dieldrin than the smaller fish. After 28 days of elimination, dieldrin levels in both size groups were nearly equal. Catfish exposed to 2 ppm (mg/kg) dieldrin through their diets accumulated significantly more dieldrin in muscle than did fish exposed to 75 pptr in water. When fish were exposed to dieldrin both in food and water, dieldrin from both sources contributed to the total dieldrin load. Large catfish accumulated more dieldrin from food and water than did smaller catfish. After 28 days of elimination, levels of dieldrin were not significantly different in muscles of 150-225 mm and 350-440 mm catfish exposed via both food and water.     

Biomarker assessment of the effects of petroleum refinery contamination on channel catfish

A series of arguments are presented that emphasize the importance of comparatively evaluating psychotherapies with appropriate pharmacotherapy and pill placebo. The lack of a pill-placebo arm has rendered moot those studies that compared pharmacotherapy directly with psychotherapy because of the lack of an internal sample defining calibration with regard to medication responsivity. The National Institute of Mental Health (NIMH) Treatment of Depression Collaborative Research Program is critically discussed because this program incorporated the recommended design features but still led to substantial controversy. The inclusion of a pill-placebo arm in future therapeutic studies is most desirable. NIMH should initiate a funded program specifically for multisite, pill-placebo-controlled studies of psychotherapy, pharmacotherapy, and their combination, jointly sponsored and supervised by skilled psychopharmacologists and psychotherapists.     

T-cell receptors in channel catfish: structure and expression of TCR alpha and beta genes

Herein are reported full length cDNA sequences for TCR alpha- and beta-chains of the channel catfish. Included are sequences belonging to four Valpha and six Vbeta families which share hallmarks in common with the Valpha and Vbeta genes of other species. Similar to the situation in other vertebrates, the catfish Calpha and Cbeta sequences exhibit distinct immunoglobulin, connecting peptide, transmembrane and cytoplasmic domains. However, the catfish TCR Calpha and Cbeta regions are shorter than those of mammals and the catfish Cbeta chain lacks a cysteine in its connecting peptide region. Two different catfish Cbeta cDNA sequences were identified, suggesting the existence of either two Cbeta loci or allotypes. Based on Southern blot analyses, each of the catfish TCR gene loci appear to be arranged in a translocon (as opposed to multicluster) organization with multiple V elements and a single or few copies of C region DNA. At the deduced amino acid level, the catfish Cbeta sequence exhibits 42% identity with the Cbeta of Atlantic salmon, 41% identity with the Cbeta of rainbow trout and 26% identity with Cbeta of the horned shark. The catfish Calpha amino acid sequence exhibits 44 and 29% identity with Calpha of the rainbow trout and southern pufferfish, respectively. TCRalpha and beta messages are selectively expressed and rearranged in a catfish clonal cell line that appears to be of the T lineage. This TCR alpha/beta expressing clonal lymphocyte line, designated 28S.1, has T-cell like function in that it constitutively produces a supernatant factor(s) with growth promoting activity. These findings should facilitate functional studies of fish TCRs and T cells in ways not previously possible with other 'lower' vertebrate models.     

Synthesis, characterization and antimicrobial activity of zinc and cerium co-doped a-zirconium phosphate

A series of zinc ions or/and cerium ions co-doped α-zirconium phosphate (Zn-Ce@ZrPs) were prepared. The novel Zn-Ce@ZrPs were characterized and the antibacterial activity on Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus were tested. The results showed that zinc ions (Zn2+) or/and cerium ions (Ce3+) were combined with ZrP, and the Ce3+ was adsorbed on the surface of ZrP through hydrogen bonds, while Zn2+ intercalated into the interlayer of ZrP. Zn-Ce@ZrPs showed excellent synergistic antibacterial activity. When Zn2+/Ce3+ atomic ratio was 0.6, the Zn-Ce@ZrP3 showed the highest synergistic antibacterial efficiency, suggesting great potential application as antibacterial agents in microbial control.