Differential expression of proliferative, cytoskeletal, and adhesive proteins during postnatal development of the hamster submandibular gland

 Although the submandibular gland (SMG) plays important exocrine and endocrine roles, little is known about the molecular details underlying its development. Previously, we reported that in the postnatally developing hamster SMG, GPT, the protein product of the first N-glycosylation gene, ALG7, was an in vivo marker for salivary cell proliferation. Here we investigated the proliferative, cytoskeletal, and adhesive changes during SMG postnatal development. The cellular localization and abundance of GPT, filamentous actin, and β1 integrin receptor were examined using confocal microscopy and immunoblotting. In neonatal glands, high GPT levels marked extensive cell proliferation throughout the tissue. The apical regions of immature salivary cells displayed intense actin staining, while most of the β1 integrin was diffusely distributed throughout the tissue. As development proceeded, discrete regions of the gland expressed attenuated levels of GPT, an increased organization of actin to the cell cortex, and β1 integrin to the basal lamina. In the adult SMG, differentiated salivary cells displayed low levels of GPT and actin. While the abundance of β1 integrin remained unchanged throughout development, in the adult, it was found exclusively in regions where cells contact the basal lamina. These data indicate that SMG development entails regionalized cell proliferation and polarization, and that these processes are temporally and spatially coordinated with the establishment of stable cell-substratum interactions. Accepted: 26 October 1998     

Antimicrobial and biological effects of N-diphenylphosphoryl-P-triphenylmonophosphazene-II and di(o-tolyl) phosphoryl-P-tri(o-tolyl)monophosphazene-III on bacterial and yeast cells

The purpose of the study was to synthesize and evaluate the antimicrobial effects of two monophosphazenes, N-diphenylphosphoryl-P-triphenylmonophosphazene-II and N-di(o-tolyl)phosphoryl-P-tri(o-tolyl)monophosphazene-III on bacterial and yeast strains. The biological effects of these molecules were compared with a potential antioxidant vitamin E. According to results, the triphenyl monophosphazene-II has antimicrobial effect on all the bacterial and yeast cells, but tri(o-tolyl)monophosphazene-III has only antimicrobial effect on some bacterial cells. When the concentration of triphenyl monophosphazene-II was raised, it was observed that inhibition zone increased on the bacterial growth media. The biological effects of these molecules were compared to vitamin E in the Saccharomyces cerevisiae culture media. In 200 microg administered culture media, the cell density decreased in vitamin E, triphenyl monophosphazene-II and tri(o-tolyl)monophosphazene-III groups at the end of 24 and 48 h incubation times (p<0.001,p<0.05). While the cell densities in vitamin E and tri(o-tolyl)monophosphazene-II groups decreased partly at the end of 72 h incubation time (p<0.05), its level in triphenyl monophosphazene-II group increased (p<0.01) at the same incubation time. In 1,000 microg administered culture media, cell density was not found to differ between vitamin E and control groups at the end of 24h incubation time, but it was found that the cell densities in triphenyl monophosphazene and tri(o-tolyl)monophosphazene-III groups decreased at the same incubation time (p<0.001). The cell densities in tri(o-tolyl)monophosphazene-III group and triphenyl monophosphazene-II decreased at the end of 48 h incubation time (respectively, p<0.05,p<0.001). In 200 microg administered cell pellets, while the lipid level was not found to differ between control and vitamin E, the lipid level decreased in triphenyl monophosphazene-II and tri(o-tolyl)monophospazene-III groups (respectively, p<0.001,p<0.01). In 1,000 microg administered cell pellets, it was found that the lipid level decreased in vitamin E, triphenyl monophosphazene-II and tri(o-tolyl)monophosphazene-III groups (p<0.001,p<0.01).     

Pharmacokinetics and biodistribution of radioimmunoconjugates of anti-CD19 antibody and single-chain Fv for treatment of human B-cell Malignancy

The comparative advantages and disadvantages of intact antibodies and single-chain Fv as immunotoxins and radioimmunoconjugates have been widely discussed but not directly compared. In this study, the in vivo properties of anti-CD19 B43 monoclonal antibody and its derived single-chain Fv (FVS191) were studied in athymic nude mice bearing CD19-positive human lymphomas. B43 mab and FVS191 were labeled with iodine-125 using iodine-beads, and immunoreactivities were determined to be 57% and 72%, respectively. Scatchard analysis showed a similar high affinity for both. The results of pharmacokinetic studies revealed that FVS191 had a rapid biphasic clearance from the circulation (T1/2α = 2.5 min, T1/2β = 3.7 h); The T1/2α and T1/2β phases of B43 mab were determined to be 0.72 h and 57 h respectively. Biodistribution studies compared the uptake of labeled antibodies by CD19-positive and by CD19-negative tumors. The peak percentages of injected dose were 5.7% at 12 h for B43 and 2.45% at 1 h for FVS191. Radiolocalization indices (RI) demonstrated tumor-specific uptake for both, but higher uptake for B43. The optimal RI was seen at 15 min for FVS191 and 6 h for B43. FVS191 was unstable in vivo, approximately 50% of the injected dose being degraded in blood in 100 min. Radioactivity detected in the urine was present mainly as the deiodinized form of FVS191. The results suggest that B43 mab is favored over FVS191 in biodistribution properties and in vivo stability. Because B43 Mab showed early tumor-specific uptake, high RI values, and favorable tissue-to-blood ratios, it is a potential candidate for radioimmunotherapy and immunotoxin therapy of B-cell leukemia and lymphoma. Received: 17 June 1997 / Accepted: 17 June 1998     

Microbiological hydrogen (H2) thresholds in anaerobic continuous-flow systems: Effects of system characteristics

Hydrogen (H₂) concentrations that were associated with microbiological respiratory processes (RPs) such as sulfate reduction and methanogenesis were quantified in continuous-flow systems (CFSs) (e.g., bioreactors, sediments). Gibbs free energy yield (ΔǴ ~ 0) of the relevant RP has been proposed to control the observed H₂ concentrations, but most of the reported values do not align with the proposed energetic trends. Alternatively, we postulate that system characteristics of each experimental design influence all system components including H₂ concentrations. To analyze this proposal, a Monod-based mathematical model was developed and used to design a gas–liquid bioreactor for hydrogenotrophic methanogenesis with Methanobacterium bryantii M.o.H. Gas-to-liquid H₂ mass transfer, microbiological H₂ consumption, biomass growth, methane formation, and Gibbs free energy yields were evaluated systematically. Combining model predictions and experimental results revealed that an initially large biomass concentration created transients during which biomass consumed [H₂]_L rapidly to the thermodynamic H₂-threshold (≤1 nM) that triggerred the microorganisms to stop H₂ oxidation. With no H₂ oxidation, continuous gas-to-liquid H₂ transfer increased [H₂]_L to a level that signaled the methanogens to resume H₂ oxidation. Thus, an oscillatory H₂-concentration profile developed between the thermodynamic H₂-threshold (≤1 nM) and a low [H₂]_L (~10 nM) that relied on the rate of gas-to-liquid H₂-transfer. The transient [H₂]_L values were too low to support biomass synthesis that could balance biomass losses through endogenous oxidation and advection; thus, biomass declined continuously and disappeared. A stable [H₂]_L (1807 nM) emerged as a result of abiotic H₂-balance between gas-to-liquid H₂ transfer and H₂ removal via advection of liquid-phase.     

Microbial transformation of (-)-carvone

The cyclic monoterpene ketone (-)-carvone was metabolized by the plant pathogenic fungus Absidia glauca. After 4 days of incubation, the diol 10-hydroxy-(+)-neodihydrocarveol was formed. The absolute configuration and structure of the crystalline substance was identified by means of X-ray diffraction and by spectroscopic techniques (MS, IR and NMR). The antimicrobial activity of the substrate and metabolite was assayed with human pathogenic microorganisms.     

Plasma values of oxidants and antioxidants in acute brain hemorrhage

The levels of oxidants xanthine oxidase (XO), nitric oxide (NO), and malondialdehyde (MDA) and of the antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and glutathione reductase (GRD) were determined in plasma within 24 h after onset of hemorrhagic stroke in 17 patients (9 men and 8 women, aged 60.7+/-11.5 yr) and in 20 healthy controls (12 men and 8 women, aged 62.5+/-8.3 yr). Compared to controls, the plasma SOD and total superoxide scavenger activities (TSSA) were significantly lower and the NO levels were significantly higher among the stroke patients. XO showed a slight, nonsignificant increase in the patients, but the levels of MDA, NSSA, GRD, and GSH-Px did not show any significant differences between the two groups. The hemorrhage volume was negatively correlated with the initial score of the Glasgow Coma Scale and a positive correlation with lethal outcome, but it did not correlate significantly with any of the measured parameters. The results suggest that free radicals might play a role in the development of brain injury following brain hemorrhage.     

Influence of smoking on serum and milk malondialdehyde, superoxide dismutase, glutathione peroxidase, and antioxidant potential levels in mothers at the postpartum seventh day

The aim of the study was to investigate simultaneously serum and milk malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione peroxidase (GPx) activities, and antioxidant potential (AOP) in active-smoking, passive-smoking, and nonsmoking mothers and to search if there is any difference between serum and milk oxidant/ antioxidant status caused by smoking. According to their smoking status, 60 mothers (age range: 20–35 yr) were classified into one of three groups: the active-smoking mothers (n=15), the passive-smoking mothers (n=22), and the nonsmoking mothers (n=23). Serum and milk MDA, SOD, GPx, and AOP values were determined in mothers on the postpartum seventh day by the spectrophotometric method. Serum Zn and Cu concentrations were determined by atomic absorption spectrophotometry (AAS). There was no significant difference in serum samples with respect to MDA (p=0.17), SOD (p=0.51) and AOP (p=0.36) levels, but there was a significant difference in serum GPx (p=0.002) levels among the study groups. The significant differences were also found in milk samples in terms of MDA (p=0.002) and SOD (p=0.011), but not in GPx (p=0.11) and AOP (p=0.29) levels among the study groups. No significant difference was seen in serum zinc concentration (p=0.49), but copper concentration differed significantly among the groups (p=0.005). These observations suggest that human milk is more vulnerable to oxidative stress and lipid peroxidation than serum samples in smoking mothers, even if they are passive smokers.