Inhalation of an Ethanol-Based Zileuton Formulation Provides a Reduction of Pulmonary Adenomas in the A/J Mouse Model

Potential efficacy of zileuton, a 5-LOX inhibitor, was evaluated for the reduction of pulmonary adenomas in the A/J murine model when administered via nose-only inhalation. Development of pulmonary adenomas was induced with benzo(a)pyrene. Animals were treated with a zileuton solution (5 mg/mL in 85:15 ethanol/water) either twice weekly or five times a week via nose-only inhalation; The placebo solution (85:15 EtOH/H₂O, no active) was also evaluated. Dose delivered was calculated to be 1.2 mg/kg per exposure for each zileuton group. After 20 weeks of treatment, surface tumors were enumerated and histologically assessed. A significant reduction in tumor count was noted for both the twice weekly administration (40%) and the five times a week administration (59%). The data also showed a significant reduction for the group, which received the placebo (approximately 58%). The treatment groups were also found to have an impact on the histological stages of adenoma development.     

Penicillin-binding proteins 4 and 5 of Haemophilus influenzae are involved in cell wall incorporation

Abstract Permeabilized cells of Haemophilus influenzae incorporate wall precursors into murein material in an ampicillin-sensitive reaction. In resistant transformants that contain the low antibiotic affinity penicillin-binding proteins (PBPs) 4 and 5, the sensitivity of this incorporation reaction to ampicillin is proportionally lower, suggesting a catalytic role for these proteins in wall synthesis. We conclude that, analogous to the reaction in Escherichia coli , PBPs 4 and 5 of H. influenzae have transpeptidase activity.     

Improved micro-method for the high-performance liquid chromatographic determination of caffeine and paraxanthine in biological fluids

A high-performance liquid chromatographic procedure is reported for reproducibly and sensitively quantitating caffeine and its N-demethylated metabolite paraxanthine in micro-samples. A 5-μm reversed-phase radial compression column and 214-nm fixed wavelength ultraviolet detector were used to attain a sensitivity sufficient to quantitate these compounds at concentratios as low as 80 ng/ml using only 25 μl of sample. The assay is applicable to microliter samples of whole blood, serum, plasma, saliva, amniotic, cerebro-spinal and gastric fluids such as might be obtained in studies involving small animals or neonates. The utility of the assay is illustrated with caffeine and paraxanthine levels measured in several maternal and fetal fluids following constant-rate intravenous infusion of caffeine into a rabbit throughout pregnancy.     

Autoantibody to vasoactive intestinal peptide in human circulation

In a radioassay for Vasoactive Intestinal Peptide (VIP)-binding, eight out of 33 plasma samples from healthy human subjects exhibited specific binding ranging from 2.6% to 46.7% of total [125 I]VIP. This binding was competitively displaced by unlabeled VIP. The structurally homologous peptides, Peptide Histidine Isoleucine (PHI) and secretin, were, respectively, 72-fold and 413-fold less potent than VIP in displacing bound [125 I]VIP, whereas the unrelated peptides, neurotensin, eledoisin, bombesin and metenkephalin, were without effect on the binding. The antibody nature of the VIP-binding factor was suggested by its precipitation with ammonium sulfate, attenuation after absorption with Staphylococcus aureus preparations, precipitation with antisera against human IgG and IgM, and coelution with standard IgG and IgM on anion-exchange and high-performance gel-filtration columns. Pepsin treatment of purified IgG fraction yielded a VIP-binding species with apparent molecular weight of 108 +/- 13 kDa that was precipitated by antiserum against the F(ab)2 fragment of the IgG molecule. These results demonstrate the existence in some human plasmas of an autoantibody that binds VIP.     

Nitroimidazole conjugates of bis(thiosemicarbazonato)Cu(II) - Potential combination agents for the PET imaging of hypoxia

Combination agents comprising two different pharmacophores with the same biological target have the potential to show additive or synergistic activity. Bis(thiosemicarbazonato)copper(II) complexes (e.g. ⁶⁴Cu-ATSM) and nitroimidazoles (e.g. ¹⁸F-MISO) are classes of tracer used for the delineation of tumor hypoxia by positron emission tomography (PET). Three nitroimidazole-bis(thiosemicarbazonato)copper(II) conjugates were produced in order to investigate their potential as combination hypoxia imaging agents. Two were derived from the known bifunctional bis(thiosemicarbazone) H₂ATSM/A and the third from the new precursor diacetyl-2-(4-N-methyl-3-thiosemicarbazone)-3-(4-N-ethylamino-3-thiosemicarbazone) - H₂ATSM/en. Oxygen-dependent uptake studies were performed using the ⁶⁴Cu radiolabelled complexes in EMT6 carcinoma cells. All the complexes displayed appreciable hypoxia selectivity, with the nitroimidazole conjugates displaying greater selectivity than a simple propyl derivative used as a control. Participation of the nitroimidazole group in the trapping mechanism is indicated by the increased hypoxic uptake of the 2- vs. the 4-substituted ⁶⁴Cu-ATSM/A derivatives. The 2-nitroimidazole derivative of ⁶⁴Cu-ATSM/en demonstrated superior hypoxia selectivity to ⁶⁴Cu-ATSM over the range of oxygen concentrations tested. Biodistribution of the radiolabelled 2-nitroimidazole conjugates was carried out in EMT6 tumor-bearing mice. The complexes showed significantly different uptake trends in comparison to each other and previously studied Cu-ATSM derivatives. Uptake of the Cu-ATSM/en conjugate in non-target organs was considerably lower than for derivatives based on Cu-ATSM/A.     

Orientation and ingestion rates of larval Anopheles albimanus in response to floating particles

Response of fourth-instar larvae of Anopheles albimanus Wiedemann (Diptera: Culicidae) to food and inert particles floating at the water surface was studied. In a choice test, larvae aggregated at powdered organic materials (blood meal, liver powder alfalfa flour and wheat flour) but not at inert materials (kaolin, chalk or charcoal). Larvae responded positively to proteins as well as some carbohydrates, but not to cellulose. Retention of larvae at food sources found by random locomotion was found to be responsible for larval aggregation. Larvae ingested food particles 6–9 times faster than insert particles. The significance of Anopheline feeding behavior in the development of formulations of stomach toxins (bacterial agents) used in larval control is discussed.

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Zusammenfassung Die vorliegende Studie befasst sich mit Verhaltensreaktionen von Anopheles albimanus Viertlarven auf an der Wasseroberfläche schwimmende Partikel. Verteilung und Orientierung der Larven wurde in einer Wahlapparatur quantifiziert. Nach Auftrag von Alfalfamehl, Weizenmehl, Stärke, Blutmehl, Leberpulver und Fischmehl wurde Aggregation der Larven in den beköderten Fächern der Apparatur beobachtet. Sowohl Proteine (Casein) als auch einige Kohlehydrate (Amylose, Amylopectin) lösten Aggregationen der Larven aus. Im Unterschied dazu führte Auftrag von Kreide, Kaolin, Polyaethylenpulver, Talcum oder Cellulose nicht zu Aggregationen. Zur Beschreibung der Entstehung larvaler Aggregationen bei Futterstoffen wurden die Schwimmbewegungen der Larven in Anwesenheit von Weizenmehl als Ködersubstanz quantifiziert. Da keine Attraktion der Larven im Sinne einer gerichteten Schwimmbewegung beobachtet werden konnte, wird geschlussfolgert, dass sofortige Beendung der Suchaktivität der Larven bei zufällig gefundenen Futterquellen für die beobachteten Aggregationen bei organischen Substanzen verantwortlich ist.Die Fressraten der Larven bei Angebot verschiedener Substanzen im Überschuss wurde bestimmt. Larven fülten drei von insgesamt sechs Darmabschnitten innerhalb von 15–30 min bei Angebot von Futtersubstanzen, während die Füllung von nur zwei Darmabschnitten mit inerten Materialien erst nach 90–120 min zu beobachten war. Die Resultate werden in Bezug auf wasseroberflächengebundene Formulierungen von Frassgiften diskutiert. Inerte Trägersubstanzen werden wahrscheinlich wesentlich langsamer aufgenommen als Futtersubstanzen. Da An. albimanus Larven nicht von Futterquellen angezogen werden, ist eine rasche und wirksame Giftaufnahme besonders dann zu erwarten, wenn die gesamte Oberfläche der Brutgewässer mit toxinhaltigen Trägerpartikeln bedeckt werden kann.