Fine Mapping the Spatial Distribution and Concentration of Unlabeled Drugs within Tissue Micro-Compartments Using Imaging Mass Spectrometry

Readouts that define the physiological distributions of drugs in tissues are an unmet challenge and at best imprecise, but are needed in order to understand both the pharmacokinetic and pharmacodynamic properties associated with efficacy. Here we demonstrate that it is feasible to follow the in vivo transport of unlabeled drugs within specific organ and tissue compartments on a platform that applies MALDI imaging mass spectrometry to tissue sections characterized with high definition histology. We have tracked and quantified the distribution of an inhaled reference compound, tiotropium, within the lungs of dosed rats, using systematic point by point MS and MS/MS sampling at 200 µm intervals. By comparing drug ion distribution patterns in adjacent tissue sections, we observed that within 15 min following exposure, tiotropium parent MS ions (mass-to-charge; m/z 392.1) and fragmented daughter MS/MS ions (m/z 170.1 and 152.1) were dispersed in a concentration gradient (80 fmol-5 pmol) away from the central airways into the lung parenchyma and pleura. These drug levels agreed well with amounts detected in lung compartments by chemical extraction. Moreover, the simultaneous global definition of molecular ion signatures localized within 2-D tissue space provides accurate assignment of ion identities within histological landmarks, providing context to dynamic biological processes occurring at sites of drug presence. Our results highlight an important emerging technology allowing specific high resolution identification of unlabeled drugs at sites of in vivo uptake and retention.     

Untersuchungen über den osmotischen Wert polyploider Pflanzen

Zusammenfassung In der Mehrzahl der untersuchten Fälle (insgesamt über 1000 Einzelmessungen) wurden die osmotischen Werte der Tetraploiden niedriger gefunden als die der Diploiden, doch sind die Unterschiede im allgemeinen gering. Junge Pflanzen hatten immer niedrigere 4n-Werte; bei alten Pflanzen und bei Pflanzen unter extremen Kulturbedingungen waren aber vielfach die Werte der 4n höher. Es wird die Hypothese aufgestellt, daß niedrigere osmotische Werte der Tetraploiden den Normalzustand darstellen, daß aber die Polyploiden wie in anderen Eigenschaften, so auch in ihrer Osmotik variabler sind und unter dem Einfluß von Außen-bedingungen ihre osmotischen Werte stärker modifizieren.Unter physiologischen Zwangsbedingungen (Trockenkultur, Pfropfungen) wurde an den Tetraploiden eine unter den gegebenen Verhältnissen zweckmäßige Erhöhung ihrer osmotischen Werte über das Niveau der 2n beobachtet. Gegen Trockenklima erwiesen sich die untersuchten Tetraploiden im Vergleich mit den Diploiden resistenter.Irgendwelche artspezifischen Eigentümlichkeiten konnten im Rahmen der untersuchten Pflanzen nicht festgestellt werden.Mit 6 Textabbildungen.     

Autophosphorylation of untrained and well-trained skeletal muscle myosin

Earlier the autophosphorylation of myosin and the labile phosphate (P) content of rabbit skeletal muscle was reported [6, 7, 9]. The present paper describes that the endogeneous preformed P level in fresh preparation of exercised muscle is higher than that of untrained control one. It was revealed that the presence of a significant amount of mitochondrial myosin (with much higher P content) in the well-trained human muscle preparations falsified the appreciation of myofibrillar myosin. Therefore, a reliable myofibrillar preparation with correct P content from exercised subjects was obtained only after the separation of mitochondrial fraction. The P content of fresh preparations can be increased by phosphorylation even in the exercised muscle myosins up to the most higher level in human samples. The phosphoryl group incorporation from [gamma-32P]ATP into the rabbit and hare myosins was checked by radioactive tracer technique, and confirmed by total P content determination performed parallel with molybdate test. It was stated that under present circumstances the labelled 32P incorporation was lower even at an optimal substrate concentration than that of P value obtained directly with molybdate method; because the total P content of preparations had not exchanged during 2 min incubation. So it has been concluded from [gamma-23P] phosphoryl group assayments that much higher amount of P was incorporated into P-Arg, N pi-P-His and fraction 2 as compared with unappreciated labelled P level of the inorganic P (P-Ser, P-Thr), P-Lys, N tau-P-His and minor fractions. From these observations it has been considered that the P-Arg, N pi-P-His and fraction 2 take part in the contraction mechanism and in the course of physical training.     

Does “ring syndrome” exist? An analysis of 207 case reports on patients with a ring autosome

Summary Analysis of 207 case reports on patients with ring autosome showed that: (1) Forty patients, a fifth of the total, had extreme growth failure together with an otherwise almost-normal appearance, viz. no major malformation, no specific deletion syndrome, no or only a few unspecific minor anomalies. This phenotype may be regarded as the ring syndrome, a term proposed by Cote et al. (1981) since it is independent of what chromosome is involved. (2) Severe growth failure, the sole major physical abnormality in the ring syndrome, was seen significantly more often among patients with ring of larger chromosomes than among patients with a smaller ring, indicating that the greater the chromosome involved in ring formation, the higher is the probability of severe growth failure. (3) Larger ring chromosomes showed significantly more often instability than smaller rings, suggesting that there may be a correlation between ring instability and the size of the chromosome involved. (4) Growth failure was present in significantly more patients with a labile ring than with a stable ring, indicating that a correlation may exist between ring instability and growth failure. It is suggested that the ring syndrome observed in many cases with ring autosome may result from end-to-end fusion of chromosome ends, an event not involving deletion in the genetic sense. It is also suggested that the ring syndrome is caused by a continuous generation of secondary aneuploid cells with increased mortality, i.e. structural ring instability which seems to be a function of the size of the chromosome involved. Thus, formation of a ring chromosome in certain cases might be regarded as a structural mutation, i.e. an alteration in the structure of the genetic material per se, rather than a loss or gain of genetic dosages.     

Cooperative binding of primycin and gramicidin on erythrocyte membranes. A cation transport study

In this paper the authors present a comparative study of the actions of the antibiotics primycin and gramicidin on the erythrocyte membrane permeability. It has been found that both antibiotics have a nonlinear effect on the membrane permeability. Above a threshold antibiotic concentration, which is characteristic of the type of the antibiotic, the cation permeability of the erythrocyte membranes increases sharply. In the range of nonlinearity the transport-kinetic curves level off before achieving the equilibrium radioactive ion distribution between the extra- and intracellular spaces. A stochastic model of the cooperative and aspecific incorporation of antibiotic molecules into the membrane explains the experimental findings. The authors conclude that membrane permeability increases at the places where two or more antibiotic molecules form aggregates in the membrane.     

Effect of glycosaminoglycans and divalent cations on the thermal properties of dipalmitoylphosphatidylcholine

The effect of glycosaminoglycans (GAG) and divalent cations on the thermal properties of dipalmitoyl-phosphatidylcholine (DPPC)-water systems was examined in order to model some interactions taking place on low density lipoprotein (LDL) surfaces. The thermal properties of these systems were measured by differential scanning calorimetry (DSC). According to the results, all three glycosaminoglycans used (chondroitin-4-sulfate, chondroitin-6-sulfate and heparin) were effective but to a different extent. Calcium ions enhance the interaction more than magnesium ions, probably because divalent cations form bridges between the negatively charged groups of GAGs and the headgroups of lipids. It is conceivable that similar processes might occur in the case of LDL.     

Morphology of neurons and axon terminals associated with descending and ascending pathways of the lateral forebrain bundle in Rana esculenta

Summary The cells of origin of afferent and efferent pathways of the lateral forebrain bundle were studied with the aid of the cobalt-filling technique. Ascending afferents originated from the lateral thalamic nucleus, central thalamic nucleus, posterior tuberculum and the cerebellar nucleus. They terminated in the anterior entopeduncular nucleus, amygdala and the striatum. Telencephalic projection neurons, which are related to the lateral forebrain bundle, were located mainly in the ventral striatum and the anterior entopeduncular nucleus, but were not so numerous in the dorsal striatum. Irrespective of their location, most of the neurons projecting axons into the lateral forebrain bundle had piriform or pyramidal perikarya. Long apical dendrites usually arborized in a narrow space, whereas widely arborizing secondary dendrites originated from short dendritic trunks. The other neurons that contributed to the lateral forebrain bundle were fusiform or multipolar cells. Striatal efferents terminated in the pretectal area and in the anterodorsal, anteroventral and posteroventral tegmental nuclei.     

Penicillin-binding proteins of protoplast and sporoplast membranes of Streptomyces griseus strains

Membrane-bound penicillin-binding proteins (PBPs) of two Streptomyces griseus strains that sporulate well in liquid and solid medium have been investigated during the course of their life-cycle. The PBP patterns were analyzed by sodium dodecylsulphate polyacrylamide-gel electrophoresis and fluorography. One strain (No. 45 H) has only a single band (mol wt: 27,000) in early log phase, and two additional PBPs of higher mol wt (69,000 and 80,000) in the late log phase. The other strain (No. 2682) possessed two bands with mol wts 27,000 and 38,000 which did not change during its vegetative phase. In strain No. 2682, a new PBP with a mol wt of 58,000 appeared in spore membranes while one of those (mol wt 38,000) present in mycelial membranes disappeared. Our results suggest that appearance of the new PBP in the spore may be associated with the sporulation process. The major PBP band (mol wt: 27,000) present in all stages of the life cycle of these strains, may be characteristic of S. griseus while the other PBPs reflect certain stages of the life cycle. A new method was developed for the production of spore protoplasts by consecutive enzymatic treatments.Abbreviation PBP penicillin-binding protein