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A new deterministic mathematical model for North American box-office film grosses is presented. The model may be simplified to a set of non-linear ordinary differential equations describing the evolution over time of the film’s gross and exhibited sites. The novel feature of this work is the inclusion of geography-based effects to model moviegoer and exhibitor behaviour. Several key regimes are identified, depending on the popularity of the film, as well as how the screens are divided among geographical regions. Analytical results are presented for several relevant cases. Numerical simulations demonstrate close agreement between the model’s predictions and actual box-office data.

This work expands the knowledge base concerning the formation of oxides of nitrogen (NO x) in the precombustion chamber (PCC) of a four-stroke lean burn (4SLB) industrial gas engine. Two analysis methods were used to characterize specific exhaust constituent concentrations formed in the PCC. The first method extracted gas from the PCC and the overall engine exhaust to be analysed by the industry standard five gas rack (THC, NO x, O 2, CO 2, and CO) and a Fourier transform infrared spectrometer (FTIR) to quantify formaldehyde (CH2O), nitric oxide (NO), and nitrogen dioxide (NO2) concentrations. This paper focuses only on NO x. The engine was operated at the manufacturer's recommended conditions and at the engine's lean limit. A PCC air-fuel ratio (A/F) map was performed at each engine operating condition by varying the fuel supply pressure to the PCC. An analytical model was developed to estimate the mass flow between the PCC and main cylinder, and mass flow out of the PCC through a sample extraction apparatus. The model was applied at specific instances during the engine cycle to clarify raw emissions data obtained directly from a cylinde's PCC. The model was developed to determine the origin of the mass sampled from the PCC which, in turn, was used to correct the measured pollutant concentrations. The corrected concentrations allowed for a comparison between NO x, emissions formed in the PCC and the overall engine exhaust on a mass specific basis. PCC-formed NO x, were found to constitute ε10 per cent while operating at the manufacture's recommended conditions and upwards of ε75 per cent when at the lean limit. The second test method utilized the high-speed chemiluminescence capabilities of a Cambustion fNOX400 analyser to measure NO concentration in gas extracted from the PCC on a half crank angle basis. The intra-cycle NO data were recorded simultaneously with pressure traces from the PCC and main cylinder by a Hi-Techniques combustion analysis system. The high-speed data acquisition system provided qualitative information on both intra-cycle NO concentrations and cycle-to-cycle variations.

Transitional cell carcinoma (TCC) is the second most common urologic malignancy, and 70% of patients present with superficial, or non-muscle invasive disease (NMIBC). Bacillus Calmette-Guerin (BCG), currently the most effective intravesical agent at preventing disease recurrence, is the only therapy shown to inhibit disease progression. Unfortunately, approximately 20% of patients discontinue BCG due to local and systemic toxicity and more than 30% show evidence of recurrence; this has led to increased interest in alternate chemotherapeutic agents. Induction intravesical chemotherapy has shown comparable efficacy to BCG in select patients and the immediate perioperative instillation of chemotherapeutic agents has become standard of care. Clinical trial evidence demonstrating the efficacy of BCG plus interferon 2B, gemcitabine and anthracyclines (doxorubicin, epirubicin, valrubicin) in patients refractory or intolerant to BCG is accumulating. Phase I trials investigating alternative agents such as apaziquone, taxanes (docetaxel, paclitaxel), and suramin are reporting promising data. Current efforts are also being directed towards optimizing the administration of existing chemotherapeutic regimens, including the use of novel modalities including hyperthermia, photodynamic therapy, magnetically targeted carriers, and liposomes. Despite recent enthusiasm for new intravesical agents, radical cystectomy remains the treatment of choice for patients with NMIBC who have failed intravesical therapy and select patients with naive T1 tumors and aggressive features. Our aim in this report is to provide a comprehensive review of contemporary intravesical therapy options for NMIBC with an emphasis on emerging agents and novel treatment modalities.

Two laboratory stocks of Anopheles minimus, each fixed for variant electromorphs of esterases, aspartate aminotransferase, hydroxyacid dehydrogenase, phosphogluconate dehydrogenase, mannose phosphate isomerase and glycerol dehydrogenase were used to assess linkage relationships between presumed gene loci controlling this variation. The two F1, which had been obtained from crossing the stocks, were backcrossed to a parental stock. Three loci controlled the esterases and one locus each of the other enzymes. Mpi is sex-linked. The rest are autosomal and suggested relationships are: Pgd 2.3% recombination from Aat and unlinked to any other loci; Est-1-33.8%-Est-3-31.5%-Est-2-21.0%-Had. Gcd is unlinked to any other locus. There was evidence of strong interaction between the X chromosome of one stock and autosomes of the other in which individuals bearing the X chromosome of the one suffered relatively greater mortality and had delayed development with respect to other genotypic classes.

This chapter will describe transgenic techniques that permit experimental manipulation of gene expression in the mouse. These techniques create specific and permanent changes to the germline of experimental mice, and these strategies have been fruitfully applied to the study of oncology, immunology, development, and more recently neurobiology and behavior. Specifically, in the area of neurobiology, modification of gene expression in the mouse has permitted direct assessment of the role of specific genes in the development of neural physiologic processes and in regulating behavior. Since this technique and its application to mouse behavior may seem far removed from the subject of human psychiatric disorders, we will first begin with a review of some basic concepts in molecular biology, followed by a brief introduction that will place the transgenic mouse strategy in the context of other genetic approaches to studying behavior.

The ability to recombine specifically and alter DNA sequences followed by techniques to transfer these sequences or even whole genes into normal and diseased cells has revolutionized medical research and ushered the clinicians of today into the age of gene therapy. We provide urologists a review of relevant background information, outline current treatment strategies and clinical trials, and delineate current challenges facing the field of gene therapy for advanced prostate cancer.We comprehensively reviewed the literature, including PubMed and recent abstract proceedings from national meetings, relevant to gene therapy and advanced prostate cancer. We selected for review literature representative of the principal scientific background for current gene therapy strategies and National Institutes of Health Recombinant DNA Advisory Committee approved clinical trials.Current prostate cancer gene therapy strategies include correcting aberrant gene expression, exploiting programmed cell death pathways, targeting critical cell biological functions, introducing toxic or cell lytic suicide genes, enhancing the immune system antitumor response and combining treatment with conventional cytotoxic chemotherapy or radiation therapy.Many challenges lie ahead for gene therapy, including improving DNA transfer efficiency to cells locally and at distant sites, enhancing levels of gene expression and overcoming immune responses that limit the time that genes are expressed. Nevertheless, despite these current challenges it is almost certain that gene therapy will be part of the urological armamentarium against prostate cancer in this century.

Dieses Kapitel wird transgene Methoden beschreiben, die es gestatten, die Expression von Genen in der Maus experimentell zu beeinflussen. Diese molekularbiologischen Verfahren erzeugen spezifische und dauerhafte Veranderungen im Erbgut von Labormausen und sind bereits mit Erfolg in der onkologischen, immunologischen und entwicklungsbiologischen Forschung sowie in jungerer Zeit auch in der neurobiologischen und Verhaltensforschung eingesetzt worden. Speziell auf dem Gebiet der Neurobiologie ist es moglich geworden, die Rolle spezifischer Gene in der Entwicklung neurophysiologischer Ablaufe und in der Steuerung von Verhalten bei der Maus durch eine Modifikation der Genexpression unmittelbar zu beurteilen.

We previously established the autochthonous transgenic adenocarcinoma mouse prostate (TRAMP) model to facilitate characterization of molecular mechanisms involved in the initiation and progression of prostate cancer. TRAMP mice display high grade prostatic intraepithelial neoplasia or well-differentiated prostate cancer by 10-12 weeks of age. To test the hypothesis that molecular events leading to androgen independence and metastasis can occur early in the natural history of prostate cancer yet remain silent until selective pressures such as androgen deprivation are applied, we have examined the consequences of castration on the initiation and progression to metastatic prostate cancer in TRAMP mice. Cohorts were castrated at 12 weeks of age and sacrificed at 18 (T12/18) or 24 (T12/24) weeks of age, and the development of primary cancer and metastatic disease was compared to noncastrated (T18 and T24) controls. Median T12/18 and T12/24 genitourinary (GU) weight was significantly less than T18 and T24, respectively. In addition, T12/24 GU weight was significantly greater than T12/18. Histological prostate tumors developed in 3 of 7 T12/18 and 8 of 10 T12/24 mice. All tumors that developed in castrated mice were poorly differentiated in contrast to 27% in noncastrated controls. Although castration significantly decreased GU tumor burden, overall progression to poorly differentiated and metastatic disease was not ultimately delayed. These results demonstrate that prostate cancer in the TRAMP model is heterogeneous with respect to androgen dependence as early as 12 weeks of age; therefore, early androgen ablation may have a variable impact on progression in an individual mouse. Further analysis of this prostate cancer model to identify specific molecular mechanisms that determine androgen sensitivity may facilitate future initiation of appropriate individualized hormonal therapy for the management of human prostate cancer.

To develop a syngeneic transplantable system to study immunotherapeutic approaches for the treatment of prostate cancer, three cell lines were established from a heterogeneous 32 week tumor of the transgenic adenocarcinoma mouse prostate (TRAMP) model. TRAMP is a transgenic line of C57BL/6 mice harboring a construct comprised of the minimal -426/+28 rat probasin promoter driving prostate-specific epithelial expression of the SV40 large T antigen. TRAMP males develop histological prostatic intraepithelial neoplasia by 8-12 weeks of age that progress to adenocarcinoma with distant metastases by 24-30 weeks of age. The three cell lines (TRAMP-C1, TRAMP-C2, and TRAMP-C3) express cytokeratin, E-cadherin, and androgen receptor by immunohistochemical analysis and do not appear to have a mutated p53. Although TRAMP-C1 and TRAMP-C2 are tumorigenic when grafted into syngeneic C57BL/6 hosts, TRAMP-C3 grows readily in vitro but does not form tumors. The T antigen oncoprotein is not expressed by the cell lines in vitro or in vivo. The rationale for establishing multiple cell lines was to isolate cells representing various stages of cellular transformation and progression to androgen-independent metastatic disease that could be manipulated in vitro and, in combination with the TRAMP model, provide a system to investigate therapeutic interventions, such as immunotherapy prior to clinical trials.

We have previously reported the development of a transgenic mouse model for prostate cancer derived from PB-Tag transgenic line 8247, henceforth designated the TRAMP (transgenic adenocarcinoma mouse prostate) model. We now describe the temporal and spatial consequences of transgene expression and report the identification and characterization of metastatic disease in the TRAMP model. TRAMP mice characteristically express the T antigen oncoprotein by 8 weeks of age and develop distinct pathology in the epithelium of the dorsolateral prostate by 10 weeks of age. Distant site metastases can be detected as early as 12 weeks of age. The common sites of metastases are the periaortic lymph nodes and lungs, with occasional metastases to the kidney, adrenal gland, and bone. By 28 weeks of age, 100% harbor metastatic prostate cancer in the lymph nodes or lungs. We have also demonstrated the loss of normal E-cadherin expression, as observed in human prostate cancer, as primary tumors become less differentiated and metastasize. The TRAMP model provides a consistent source of primary and metastatic tumors for histopathobiological and molecular analysis to further define the earliest molecular events involved in the genesis, progression, and metastasis of prostate cancer.

PCR was used to isolate identical partial cDNA clones encoding a serotonin 5-HT 3 receptor subunit from rat nodose and superior cervical ganglia. The amino acid sequence predicted from these clones, extending from the putative transmembrane domain I to the stop codon, demonstrated a 93% homology with the 5-HT 3 receptor A (R-A) subunit cloned from NCB 20 hybridoma mouse neuroblastoma/Chinese hamster embryonic brain cells. Comparison of the sequences of the rat gene and cDNA encoding this subunit revealed a five amino acid deletion, GSLLP, located within the putative second intracellular loop of the receptor subunit. This deletion was shown to occur at an intron/exon junction. Therefore, alternative splicing was probably responsible for the presence of short (5-HT 3 R-A s ) and long (5-HT 3 R-A L ) forms of 5-HT 3 R-A mRNA in these ganglia. PCR experiments, with specific primers located upstream and downstream of the GSLLP deletion, were used to detect reverse transcribed 5-HT 3 R-A mRNAs. A short fragment (92 bp), corresponding to the deleted form, and a long fragment (107 bp), corresponding to the nondeleted form, were amplified from various regions of the CNS and peripheral ganglia of the rat, as well as from NG108-15 hybridoma cells. In the adult rat, the ratio of the two forms varied very little from one tissue to another, the long form corresponding to only ∼10% of the total 5-HT 3 R-A mRNA. Study of their respective distributions during ontogeny demonstrated a differential expression of the short and long forms in some tissues during late embryonic development, at embryonic day 17(E17) or E20. In particular, the long form amounted to about one-third of the total 5-HT 3 R-A mRNA in the cerebral cortex and hippocampus at E17, and this proportion reached 50 and 75% in the superior cervical ganglion and nodose ganglion, respectively, at E20. These data indicate that alternative splicing of the 5-HT 3 R-A mRNA is regulated in the CNS and PNS during development in the rat.

Despite a negative metastatic evaluation, up to 100% of patients with high-grade tumors are found to harbor metastatic disease within the pelvic lymph nodes at the time of surgery. This article reviews the literature regarding efforts to refine noninvasive preoperative staging through the use of serum tumor markers, improved radiologic imaging, and histologic analysis of the biopsy specimens. A discussion of the renewed controversy regarding the role of pelvic lymphadenectomy in the management of adenocarcinoma of the prostate follows.

The portion of the complementary DNA encoding the third intracellular loop of the rat 5-hydroxytryptamine 1A (serotonin) receptor was subcloned into the vector pGEX-KG and expressed in Escherichia coli as a fusion protein coupled with the glutathione S-transferase of Schistosoma japonicum . The fusion protein was purified on a glutathione-agarose affinity column and used to immunize rabbits for the production of polyclonal anti-5-hydroxytryptamine 1A receptor antibodies. Enzyme-linked immunosorbent assay revealed that antibodies were produced as early as one month after the first injection of the fusion protein, and immune response plateaued at a maximum after the third (monthly) booster injection. These antibodies only marginally affected the specific binding of [ 3 H]8-hydroxy-2-(di- n-propyl-amino ) tetralin to solubilized and membrane bound 5-hydroxytryptamine 1A receptors, and did not interfere with serotonin-induced inhibition of forskolin-stimulated adenylate cyclase negatively coupled to 5-hydroxytryptamine 1A receptors in rat hippocampal membranes. However, antibodies were able to immunoprecipitate 5-hydroxytryptamine 1A receptor binding sites solubilized from rat hippocampal membranes. The distribution of immunoautoradiographic labelling and immunohistochemical staining of rat brain sections exposed to the antibodies raised against the fusion protein superimposed to that of 5-hydroxytryptamine 1A receptor binding sites labelled by specific radioligands, with marked enrichment in the limbic areas (dentate gyrus and CA1 area in the hippocampus, lateral septum, entorhinal cortex) and the anterior raphe nuclei. The differential cellular location of immunoreactivity within the hippocampus (where dendritic fields but not pyramidal cell somas were immunostained) and the median raphe nucleus (where the plasmic membrane of somas was strongly immunoreactive) suggests that the addressing of 5-hydroxytryptamine 1A receptors might differ from one neuronal cell type to another.

An in vivo study of cisplatin (CDDP) and 5-fluorouracil (5FU) cytotoxicity was performed using a multidose matrix with a human bladder transitional cell carcinoma xenograft tumor line (DU4284) tested by subrenal capsule assay in 154 nude mice (NM-SRCA). Statistical analysis of initial growth inhibition at 20 days and host survival demonstrates therapeutic, cooperative interaction. Toxic doses of either CDDP or 5FU alone as well as low-dose combinations provided modest or no survival benefit. The single dose of CDDP (7 mg/kg) and of 5FU (100 mg/kg) was best (by analysis of efficacy and toxicity) of those tested and caused97% initial regression. While 94% of controls incurred tumor deaths by 225 days, 75% treated at this dose were tumor free and likely cured. Our conclusions were: (a) NM-SRCA human xenograft testing is excellent for rapid in vivo screening of promising treatment strategies to evaluate for efficacy at acceptable toxicity, but confirmation of true therapeutic impact should be sought by correlating initial growth inhibition with host survival; (b) enhanced survival seen only when CDDP/5FU are used together (versus either single agent) supports the value of pursuing histiotype-specific screening of potentially synergistic drug combinations; and (c) of clinical relevance, human transitional cell carcinoma is now identified as a histiotype in which a therapeutic, cooperative interaction between CDDP/5FU has been demonstrated in vivo.

Anti-5-HT receptor antibodies were raised by injecting rabbits with either a synthetic peptide corresponding to the aa 243-268 portion of the third intracellular loop of the rat 5-HTAreceptor, or a fusion protein made of the complete loop (aa 216-350) coupled to glutathione-S-transferase. Immunohistochemical staining with these antibodies allowed the visualization of 5-HTAreceptors mainly in the limbic areas and anterior raphe nuclei, as expected from previous autoradiographic data with selective 5-HTAreceptor radioligands. Double immunostainingwith anti-5-HT,, receptor antibodies and anti-5-HT or anti-choline acetyl-transferase antibodies demonstrated that these receptors are located in the plasma membrane of practically all serotoninergic neurones in anterior raphe nuclei and a small fraction of cholinergic neurones in the median septum.

Publisher Summary This chapter describes the physicochemical characteristics of the 5-HT3 receptor and the different methods used to purify the 5-HT3 receptor. The successful solubilization of 5-HT3 receptor-binding sites can be achieved from an abundant source using a large spectrum of detergents and salt conditions. However, physicochemical studies require more restrictive conditions. The successful solubilization of membrane-bound receptor proteins requires an abundant source of receptors. The chapter reviews the properties of the most common biological detergents and highlights their respective advantages and disadvantages. The solubilization of the 5-HT3 receptor-binding sites allows the determination of several of their physical properties and leads to the purification of the ligand-binding protein(s). Proteins are estimated using the bicinchoninic acid (BCA) protein assay reagent with BSA as a standard. The physicochemical properties of amino acid sequence of the 5-HT3 receptor-binding subunit are altogether consistent with 5-HT3 receptor being a member of the superfamily of ionophoric receptors.

e16086 Background: Docetaxel (D) IV every 21 days, is the only cytotoxic agent that prolongs survival in men with castrate resistant prostate cancer (CRPC). Imatinib (I), a tyrosine kinase inhibitor, modulates PDGFR-ß in tumor vasculature. Based on phase I data from our institution, we hypothesized that D plus I would prolong time to progression (TTP) in patients (pts) with CRPC. Methods: Subjects with CRPC received D 60 mg/m2 IV every 21 days plus I 400 mg PO daily. After 10 pts, the study treatment was modified due to toxicity so that pts received I 400 mg on 10 of 21 days/cycle. The primary endpoint was TTP. Secondary endpoints were rate of PSA response and overall survival (OS). The sample size of 43 pts was designed to provide 90% power to detect an increase in TTP from 5 to 8 months. Results: 43 pts enrolled from 8/05 to 9/08. Age at enrollment ranged from 54–86 years (median 69 years). 14 pts received 2 cycles of chemotherapy (mean 4.6). 12 pts had PR (41.4%), 9 had SD (31.0%), and 8 had no response (27.6%) by PSA. No objective responses were seen by CT imaging among 10 pts with measurable disease. 3 pts remain on trial. For evaluable pts, overall median TTP was 6.4 months (95% CL: 4.8, 8.4 months) compared with TTP of 5 months seen in previous trials. 23 (79%) pts had PSA progression, 3 pts died before progression, and 3 pts remain on trial. For all evaluable pts who had PR or SD by PSA (N = 21), median TTP was 7.1 months (95% CL: 5.5, 9.1 months). Median OS was 23.1 months (95% CL: 11.61 months, NR), compared with 18.9 months for GC Conclusions: Docetaxel on day 1 plus imatinib 10 days of each 21-day cycle resulted in meaningful improvement in TTP in the subset of pts who showed a response. Toxicity precludes its use in the general population, although its role in select pts with good performance status needs to be explored. [Table: see text]

A high-sensitivity, laser-excited confocal fluorescence gel scanner has been developed and applied to the detection of fluorescently labeled DNA. An argon ion laser (1-10 mW at 488 nm) is focused in the gel with a high-numerical aperture microscope objective. The laser-excited fluorescence is gathered by the objective and focused on a confocal spatial filter, followed by a spectral filter and photodetector. The gel is placed on a computer-controlled scan stage, and the scanned image of the gel fluorescence is stored and analyzed in a computer. This scanner has been used to detect DNA separated on sequencing gels, agarose mapping gels and pulsed field gels. Sanger sequencing gels were run on M13mp18 DNA using a fluoresceinated primer. The 400-microns-thick gels, loaded with 30 fmol of DNA fragments in 3-mm lanes, were scanned at 78-microns resolution. The high resolution of our scanner coupled with image processing allows us to read up to approximately 300 bases in four adjacent sequencing lanes. The minimum band size that could be detected and read was approximately 200 microns. This instrument has a limiting detection sensitivity of approximately 10 amol of fluorescein-labeled DNA in a 1 x 3-mm band. In applications to agarose mapping gels, we have exploited the fact that DNA can be prestained with ethidium homodimer, followed by electrophoresis and fluorescence detection to achieve picogram sensitivity. We have also developed methods using both ethidium homodimer and thiazole orange staining which permit two-color detection of DNA in one lane.(ABSTRACT TRUNCATED AT 250 WORDS)

Field tests were conducted to compare the effectiveness of 2 repellent formulations of N,N-diethyl-3-methylbenzamide (deet) in combination with permethrin-impregnated military uniforms against Culex sitiens and Aedes vigilax in Thailand. Repellency was determined during a 2 h crepuscular period using volunteers who had been treated with repellents 6, 8, 10, and 12 h prior to the end of each test period. An extended-duration repellent formulation (EDRF) containing 35% deet repelled significantly more Ae. vigilax than 75% deet in ethanol. Although not statistically significant, the EDRF also resulted in fewer biting attempts by Cx. sitiens. Neither formulation provided complete protection against either species 4-12 h post-application, but both provided greater overall protection against Ae. vigilax. Volunteers who wore treated uniforms without repellents were attacked by significantly fewer mosquitoes than controls.

Monoclonal antibodies were used to demonstrate the expression of four distinct metacyclic (infective insect form) trypanosome antigens on blood forms of T. rhodesiense. Metacyclic antigens were consistently expressed on the blood forms on days 4 and 5 of the first parasitemia after metacyclic infection of C57BL/6 mice. In different mice examined, the percent of blood forms expressing metacyclic antigens ranged from 46 to 85%. Immunization with irradiated day-5 blood form trypanosomes was protective against metacyclic challenge, indicating that all antigen specificities relevant to protective immunization against metacyclic challenge are expressed on blood form trypanosomes. Blood forms, in contrast to metacyclic forms, can be isolated in quantities sufficient for purification of antigens and genetic cloning.

Synechocystis 6701 phycobilisomes consist of a core of three cylindrical elements in an equilateral array from which extend in a fanlike manner six rods, each made up of three to four stacked disks. Previous studies (see Gingrich, J. C., L. K. Blaha, and A. N. Glazer, 1982. J. Cell Biol. 92:261-268) have shown that the rods consist of four disk-shaped complexes of biliproteins with "linker" polypeptides of 27-, 33.5-, 31.5-, and 30.5-kdaltons, listed in order starting with the disk proximal to the core: phycocyanin (alpha beta)6-27 kdalton, phycocyanin (alpha beta)6-33.5 kdalton, phycoerythrin (alpha beta)6-31.5 kdalton, phycoerythrin (alpha beta)6-30.5 kdalton, where alpha beta is the monomer of the biliprotein. Phycoerythrin complexes of the 31.5- and 30.5-kdalton polypeptides were isolated in low salt. In 0.05 M K-phosphate-1 mM EDTA at pH 7.0, these complexes had the average composition (alpha beta)2-31.5 and (alpha beta)-30.5 kdalton polypeptide, respectively. Peptide mapping of purified 31.5- and 30.5-kdalton polypeptides showed that they differed significantly in primary structure. In 0.65 M Na-K-phosphate at pH 8, these phycoerythrin complexes formed rods of stacked disks of composition (alpha beta)6-31.5 or (alpha beta)6-30.5 kdaltons. For the (alpha beta)-30.5 kdalton complex, the yield of rod assemblies was variable and the self-association of free phycoerythrin to smaller aggregates was an important competing reaction. Complementation experiments were performed with incomplete phycobilisomes from Synechocystis 6701 mutant strain CM25. These phycobilisomes are totally lacking in phycoerythrin and the 31.5- and 30.5-kdalton polypeptides, but have no other apparent structural defects. In high phosphate at pH 8, the phycoerythrin-31.5-kdalton complex formed disk assemblies at the end of the rod substructures of CM25 phycobilisomes whereas no interaction with the phycoerythrin-30.5 kdalton complex was detected. In mixtures of both the phycoerythrin-31.5 and -30.5 kdalton complexes with CM25 phycobilisomes, both complexes were incorporated at the distal ends of the rod substructures. The efficiency of energy transfer from the added phycoerythrin in complemented phycobilisomes was approximately 96%. The results show that the ordered assembly of phycoerythrin complexes seen in phycobilisomes is reproduced in the in vitro assembly process.

The phycobilisomes of the unicellular cyanobacterium Synechocystis 6701, grown in white light, contain C-phycoerythrin, C-phycocyanin, and allophycocyanin in a molar ration of approximately 2:2:1, and in addition, polypeptides of 99, 46, 33.5, 31.5, 30.5, and 27 x 10(3) Daltons, as well as a trace of a approximately 9 x 10(3)-dalton component. Two nitrosoguanidine-induced mutants of this organism produce aberrant phycobilisomes. Crude cell extracts of these mutants, 6701-NTG25 and NTG31, contain phycoerythrin, phycocyanin, and allophycocyanin in a molar ration of 1:5:1:1 and 0.55:0.3:1.0, respectively. The phycobilisomes from both mutants lack the 33.5 x 10(3)-dalton polypeptide. Wile-type phycobilisomes consist of a core composed of an equilateral array of three cylindrical elements surrounded by six rods in a fanlike arrangement. The rods are made up of stacked disks, 11 nm in diameter and 6 nm thick. In phycobilisomes of mutant 6701-NTG25, numerous particles with fewer than six rods are seen. Mutant 6701-NTG31 produces incomplete structures that extend from triangular core particles, through cores with one or two attached rods, to cores with as many as five rods. The structure of the core appears unaltered throughout. The amount of phycocyanin (relative to allophycocyanin) appears to determine the number of rods per core. A common assembly form seen in 6701-NTG31 is the core with two rods attached at opposite sides. From observations of this form, it is concluded that the core elements are cylindrical, with a height of 14 nm and a diameter of 11 nm. No consistently recognizable structural details are evident within the core elements.

The polypeptide product of the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase from Euglena gracilis based on the DNA sequence of the chloroplast-encoded gene is described. The large subunit polypeptide of 475 codons is co-linear with the homologous polypeptides from other chloroplasts and cyanobacteria. The amino acid sequence is 92% homologous to that of Chlamydomonas, 84% homologous to spinach, 82% homologous to maize, and 80% homologous to that of the cyanobacterium Anabaena variabilis. Known functional domains of the protein are coded by the larger exons of the gene. Introns in the gene generally occur at coding sequences specifying hydrophilic, presumably surface exposed, regions of the polypeptide. The location of some of the introns may reflect a separation of functional domains. The 5'- and 3'-ends of the rbcL transcript were determined by primer extension sequencing using reverse transcriptase and S1 nuclease protection, respectively. The transcribed but untranslated sequences are quite distinct from those from other rbcL loci.

Laboratory tests were conducted to evaluate the influence of several variables on infection rates of Trypanosama brucei rhodesiense in Glossina morsitans , viz., postinfection temperature, clone/isolate of parasite, age and sex of tsetses, and the presence or absence of serum in the infective blood meal. Flies evaluated histologically had infection rates of 10.9% in the midgut (MG), 6.9% in the proventriculus (PV), and 1.9% in the salivary glands (SG); flies assessed by dissection methods yielded MG, PV and SG infection rates of 24.3, 19.0, and 4.8%, respectively. No consistent differences were found in infection rates using the different isolates/clone of T. b. rhodesiense , nor were there differences between flies held at different postinfection temperatures (range limits: 15–21 °C). Analysis of the influence of age and sex indicated an interaction effect where males

The nucleotide sequence of 6225 base pairs (bp) of Euglena gracilis chloroplast DNA including the complete DNA sequence of the chloroplast-encoded ribulose-1,5-bisphosphate carboxylase large subunit gene along with the flanking DNA sequences is presented. The gene is greater than 5.5 kilobase pairs in length and is organized as 10 exons coding for 475 amino acids, separated by 9 introns. The exons range in size from 45 to 438 bp, while the introns range in size from 382 to 568 bp. The introns have highly conserved boundary sequences with the consensus, 5'-N GTGTGGATTT...(intron)...TTAATTTTAT N-3'. The introns are 82-85 mol% AT, with a pronounced T greater than A greater than G greater than C base bias in the RNA-like strand. They do not appear to encode any polypeptides. In addition, the introns have a conserved sequence 30-50 bp from their 3'-ends with the consensus, 5'-TACAGTTTGAAAATGA-3'. The 5'-TACA sequence bears some homology to the 5'-end of the TACTAACA sequence found in a similar location in yeast nuclear mRNA introns. The conserved sequences of the Euglena rbcL introns may be indicative of a splicing mechanism similar to that of eucaryotic nuclear mRNA introns and group II mitochondrial introns.

Immature salivary gland (SG) infections averaging 10(3) parasites per fly can apparently develop into mature gland infections averaging 10(5) parasites per fly in as little as 4 days. Frequently flies which extrude parasites in their saliva prove to have no parasites in the SG, but often show trypanosomes in the esophagus, cibarial pump, and proboscis. In some instances, SG infections have cleared, resulting in a loss of infectivity. Results of studying numbers of parasites regurgitated upon feeding or probing have shown that number to be highly variable and not necessarily related either to previous feeding status or the total number of parasites in the glands. Cloning of metacyclics in mice has been achieved, indicating that the minimum effective dose is one parasite. To date, no infections in mice have resulted from inoculation of extraglandular parasites. Histological and dissection studies support both the classical route and an alternate route of infection development in flies. No SG-infected flies have been found which did not also have proventricular and anterior and posterior midgut (AMG and PMG) infections. Although the AMG is where the heaviest MG infections occur, the PMG seems to support the last survivors in a moribund MG infection. No parasites have been found in the hindgut.

Synechocystis 6701 phycobilisomes contain phycoerythrin, phycocyanin, and allophycocyanin in a molar ratio of approximately 2:2:1, and other polypeptides of 99-, 46-, 33.5-, 31.5-, 30.5-, and 27-kdaltons. Wild-type phycobilisomes consist of a core of three cylindrical elements in an equilateral array surrounded by a fanlike array of six rods each made up of 3-4 stacked disks. Twelve nitrosoguanidine-induced mutants were isolated which produced phycobilisomes containing between 0 and 53% of the wild-type level of phycoerythrin and grossly altered levels of the 30.5- and 31.5-kdalton polypeptides. Assembly defects in these mutant particles were shown to be limited to the phycoerythrin portions of the rod substructures of the phycobilisome. Quantitative analysis of phycobilisomes from wild-type and mutant cells, grown either in white light or chromatically adapted to red light, indicated a molar ratio of the 30.5- and 31.5-kdalton polypeptides to phycoerythrin of 1:6, i.e., one 30.5- or one 31.5-kdaltons polypeptide per (alpha beta)6 phycoerythrin hexamer. Presence of the phycoerythrin-31.5-kdalton complex in phycobilisomes did not require the presence of the 30.5-kdalton polypeptide. The converse situation was not observed. These and earlier studies (R. C. Williams, J. C. Gingrich, and A. N. Glazer. 1980. J. Cell Biol. 85:558-566) show that the average rod in wild type Synechocystis 6701 phycobilisomes consists of four stacked disk-shaped complexes: phycocyanin (alpha beta)6-27 kdalton, phycocyanin (alpha beta)6-33.5 kdalton, phycoerythrin (alpha beta)6-31.5 kdalton, and phycoerythrin-30.5 kdalton, listed in order starting with the disk proximal to the core.

Moisture was the most important factor influencing the emergence of common bermudagrass (Cynodon dactylon (L.) Pers.) and weeping lovegrass (Eragrostis curvula (Schrad.) Nees.) in a clay soil over the range of bulk density and soil water pressure investigated. Both grasses exhibited approximately the same emergence at the −1/3 and −1-bar soil water pressures. Of those soil strengths and oxygen diffusion rates encountered, none limited seedling emergence of either grass. Small correlation was found between soil strength and seedling emergence.

In order to understand better the interacting variables of the radioisotope fuel properties and the operating conditions for thermionic converters a parametric study was made of the heat source and system variables. Fuel bodies and converters of various geometric shapes were studied with planar, cylindrical, and spherical emitter surfaces. The problems relating to radiological safety and shielding of the isotope fuel were reviewed. Cost and availability information was obtained for some of the more promising isotope fuels. The results of this study show that there is no completely ideal radioisotope heat source for thermionic space power systems and that the more promising isotope fuels will require important compromises between system specific weight, radiation shielding, system efficiency, and isotope containment to produce usable space power systems.

In 1985, procurements by the Army and Air Force of automated engineering storage and retrieval systems are underway: DSREDS, the Army system (Digital Storage and Retrieval Engineering Data System), and EDCARS, the Air Force system (Engineering Data Computer Assisted Retrieval System). ATT Alpharel is the major subcontractor. DSREDS and EDCARS establish complete drawing, storage and retrieval systems for these sevices and will facilitate the distribution of bid packages. The first installation will be at MICOM at the Red Stone Arsenal in Huntsville, Alabama, and the second at SM/ALC at McClellan Air Force Base in Sacramento, California. Ten other installations will follow. The program includes twelve sites, each requiring approximately 25 terminals, the capacity to store 400 gigabytes, and 400 gigabyte backup, with aperture card scanners, page scanners, large scale scanners, and computer output microfilm (COM) and hardcopy capabilities. Because the DSREDS/EDCARS system is so significant and embodies all attributes of an outstanding automated drawing system, a brief description of it will be given here.

A one-year study of Japanese encephalitis (JE) in a small focus of transmission was conducted in suburban Bangkok in 1985. Monthly data were collected on weather, vector density, sentinel pig and chick JE antibody seroconversions, and epidemiology as related to human JE cases. The primary vector species were found to be Culex gelidus and Culex tritaeniorhynchus; from which one isolate each was obtained in March and June, respectively. Pig JE antibody seroconversion peaked in April (the hottest month), with secondary peaks following in July and December. Chick seroconversions were found only in June and July. Human cases (7) in the primary focus occurred from May-July, and started 2 months following the finding of the first JEV isolate in mosquitoes and 1 month following mass JEV seroconversion in pigs. Overall, the attack rate in the focus (0.83/10(5] was greater than 4 times that of the rest of Bangkok (0.19/10(5]. Attack rates were highest in 0-9 and 10-19 year-old groups, respectively. Indications are that JEV is transmitted to humans in Bangkok at least 10 out of 12 months per year, but that cases are concentrated in the May to July period.

Anopheline mosquitoes and their relation to malaria transmission were studied during the months of March, May and July in the Karen village of Mae Tha Waw located in the northwestern mountains of Tak Province. Thirteen species were captured on human bait during 80 man-nights of collecting. Four additional species were collected during routine larval surveys. Anopheles minimus and An. maculatus comprised 92.5% of the specimens captured biting man. Anopheles minimus and An. nivipes were implicated as vectors based on the detection of sporozoite infections using enzyme-linked immunosorbent assays for Plasmodium falciparum and P. vivax. Anopheles dirus was rarely encountered and probably played little part in transmission in Mae Tha Waw during the period of study. Information is provided on nightly biting activity, incidence of disease, infectivity and larval bionomics.

Starved mature male tsetse flies (21 to 25 days old) are capable of developing salivary gland (SG) infections of Trypanosoma brucei rhodesiense at rates nearly comparable to teneral males less than 24 hours old when given an infective meal containing parasites, horse red cells and culture medium. Although the over-all SG infection rate for mature males starved for three, four or five days before infection was about half that for teneral males less than 48 hours old (8.0% v. 15.6%), males starved for four days developed infection rates (12.3%) that were comparable to those of teneral flies less than 24 hours old (11.8%). It is suggested that the acquisition of infection by mature flies should be considered when evaluating factors contributing either to maintenance of endemic infections or perhaps even epidemic infections of human sleeping sickness.

The nucleotide sequence of 6225 base pairs (bp) of Euglena gracilis chloroplast DNA including the complete DNA sequence of the chloroplast-encoded ribulose-1,5-bisphosphate carboxylase large subunit gene along with the flanking DNA sequences is presented. The gene is greater than 5.5 kilobase pairs in length and is organized as 10 exons coding for 475 amino acids, separated by 9 introns. The exons range in size from 45 to 438 bp, while the introns range in size from 382 to 568 bp. The introns have highly conserved boundary sequences with the consensus, 5'-N GTGTGGATTT...(intron)...TTAATTTTAT N-3'. The introns are 82-85 mol% AT, with a pronounced T greater than A greater than G greater than C base bias in the RNA-like strand. They do not appear to encode any polypeptides. In addition, the introns have a conserved sequence 30-50 bp from their 3'-ends with the consensus, 5'-TACAGTTTGAAAATGA-3'. The 5'-TACA sequence bears some homology to the 5'-end of the TACTAACA sequence found in a similar location in yeast nuclear mRNA introns. The conserved sequences of the Euglena rbcL introns may be indicative of a splicing mechanism similar to that of eucaryotic nuclear mRNA introns and group II mitochondrial introns.

Application of a slight osmotic stress to only a portion of young wheat roots significantly affected certain metabolic fractions throughout the entire plant. These components change rapidly with time and therefore time studies must be used in experiments of this type concerned with an evaluation of changes in metabolic fractions in plants that are induced by environmental factors.