The NH2-terminal and COOH-terminal fragments of dentin matrix protein 1 (DMP1) localize differently in the compartments of dentin and growth plate of bone

Multiple studies have shown that dentin matrix protein 1 (DMP1) is essential for bone and dentin mineralization. After post-translational proteolytic cleavage, DMP1 exists within the extracellular matrix of bone and dentin as an NH2-terminal fragment, a COOH-terminal fragment, and the proteoglycan form of the NH2-terminal fragment (DMP1-PG). To begin to assess the biological function of each fragment, we evaluated the distribution of both fragments in the rat tooth and bone using antibodies specific to the NH2-terminal and COOH-terminal regions of DMP1 and confocal microscopy. In rat first molar organs, the NH2-terminal fragment localized to predentin, whereas the COOH-terminal fragment was mainly restricted to mineralized dentin. In the growth plate of bone, the NH2-terminal fragment appeared in the proliferation and hypertrophic zones, whereas the COOH-terminal fragment occupied the ossification zone. Forster resonance energy transfer analysis showed colocalization of both fragments of DMP1 in odontoblasts and predentin, as well as hypertrophic chondrocytes within the growth plates of bone. The biochemical analysis of bovine teeth showed that predentin is rich in DMP1-PG, whereas mineralized dentin primarily contains the COOH-terminal fragment. We conclude that the differential patterns of expression of NH2-terminal and COOH-terminal fragments of DMP1 reflect their potentially distinct roles in the biomineralization of dentin and bone matrices.     

Measurement of pulsatile insulin secretion in the rat: direct sampling from the hepatic portal vein

It has previously been shown that insulin is secreted in discrete secretory bursts by sampling directly from the portal vein in the dog and humans. Deficient pulsatile insulin secretion is the basis for impaired insulin secretion in type 2 diabetes. However, while novel genetically modified disease models of diabetes are being developed in rodents, no validated method for quantifying pulsatile insulin secretion has been established for rodents. To address this we 1) developed a novel rat model with chronically implanted portal vein catheters, 2) established the parameters to permit deconvolution of portal vein insulin concentrations profiles to measure insulin secretion and resolve its pulsatile components, and 3) measured total and pulsatile insulin secretion compared with that in the dog, the species in which this sampling and deconvolution approach was validated for quantifying pulsatile insulin secretion. In rats, portal vein catheter patency and function were maintained for periods up to 2-3 wk with no postoperative complications such as catheter tract infection. Rat portal vein insulin concentration profiles in the fasting state revealed distinct insulin oscillations with a periodicity of approximately 5 min and an amplitude of up to 600 pmol/l, which was remarkably similar to that in the dogs and in humans. Deconvolution analysis of portal vein insulin concentrations revealed that the majority of insulin ( approximately 70%) in the rat is secreted in distinct insulin pulses occurring at approximately 5-min intervals. This model therefore permits direct accurate measurements of pulsatile insulin secretion in a relatively inexpensive animal. With increased introduction of genetically modified rat models will be an important tool in elucidating the underlying mechanisms of impaired pulsatile insulin secretion in diabetes.     

Multiphasic approach reveals genetic diversity of environmental and patient isolates of Mycobacterium mucogenicum and Mycobacterium phocaicum associated with an outbreak of bacteremias at a Texas hospital

Between March and May 2006, a Texas hospital identified five Mycobacterium mucogenicum bloodstream infections among hospitalized oncology patients using fluorescence high-performance liquid chromatography analysis of mycolic acids. Isolates from blood cultures were compared to 16 isolates from environmental sites or water associated with this ward. These isolates were further characterized by hsp65, 16S rRNA, and rpoB gene sequencing, hsp65 PCR restriction analysis, and molecular typing methods, including repetitive element PCR, random amplified polymorphic DNA PCR, and pulsed-field gel electrophoresis (PFGE) of large restriction fragments. Three of five patient isolates were confirmed as M. mucogenicum and were in a single cluster as determined by all identification and typing methods. The remaining two patient isolates were identified as different strains of Mycobacterium phocaicum by rpoB sequence analysis. One of these matched an environmental isolate from a swab of a hand shower in the patient's room, while none of the clinical isolates of M. mucogenicum matched environmental strains. Among the other 15 environmental isolates, 11 were identified as M. mucogenicum and 4 as M. phocaicum strains, all of which were unrelated by typing methods. Although the 16S rRNA gene sequences matched for all 14 M. mucogenicum isolates, there were two each of the hsp65 and rpoB sequevars, seven PCR typing patterns, and 12 PFGE patterns. Among the seven M. phocaicum isolates were three 16S rRNA sequevars, two hsp65 sequevars, two rpoB sequevars, six PCR typing patterns, and six PFGE patterns. This outbreak represents the first case of catheter-associated bacteremia caused by M. phocaicum and the first report of clinical isolates from a U.S. hospital. The investigation highlights important differences in the available typing methods for mycobacteria and demonstrates the genetic diversity of these organisms even within narrow confines of time and space.     

Wild plant seed storage at Neolithic Çatalhöyük East,Turkey

Full analysis of eight seed samples collected in the 1960's excavations at Neolithic Çatalhöyük East, Turkey, is presented. Detailed investigation of the composition and context of the samples suggests that the Neolithic population collected, processed and stored seeds from Capsella sp. and Descurainia sp. (wild crucifers) for food use. In addition seeds of Vicia/Lathyrus sp. (wild vetch), Helianthemum spp. and Taeniatherum caput-medusae mixed with Eremopyrum type (grasses) were also found, some of which may have been used for food or other purposes. The analysis demonstrates that wild seed exploitation was a regular part of subsistence practice alongside the economic staple of crop production, and again demonstrates how diverse plant use practices were at the site.     

Perch height predicts dominance rank in birds

Dominant individuals within animal groups will frequently place themselves in the most beneficial position for maximal protection against predation. Higher perches are generally associated with reduced predation risk in birds, so we predicted that dominant birds will preferentially place themselves on higher perches. We tested this by determining the dominance hierarchy in two populations of captive birds (Homing Pigeons Columba livia and Great Cormorants Phalacrocorax carbo), and relating rank within the dominance hierarchy to observed perch height preferences. We found that perch choice was significantly repeatable in pigeons, and that more dominant individuals of both species selected higher perches. As well as facilitating early detection of and escape from potential predators, higher perches are also likely to facilitate the display of aggression to other group members.     

The fluidity and organization of mitochondrial membrane lipids of the brown adipose tissue of cold-adapted rats and hamsters as determined by nitroxide spin probes.

A detailed study of lipid fluidity and organization in the mitochondria of the brown adipose tissue from warm- and cold-adapted rats (nonhibernators) and hamsters (hibernators) is made in order to delineate any relationship between lipid properties and the ability to lower body temperature after cold-adaptation. Complete phospholipid analyses are presented; the data are very similar for cold- and warm-adapted rats, and for cold- and warm-adapted hamsters, but the rat lipids have a higher degree of unsaturation than those of the hamsters. Spin probe analogs of stearic acid and cholestane were used to investigate at the molecular level the fluidity and order of the mitochondrial lipids. Studies were made on intact mitochondria, and in liposomes and oriented multibilayers of extracted lipids. In no case was evidence found for a phase transition in the lipids, or for a relationship between the lipid fluidity in brown adipose tissue mitochondria and the ability to survive at lowered body temperatures. The spin probes generally had a decreased mobility in mitochondria relative to extracted lipids. The electron spin resonance spectra were analyzed to include order- and time-dependent phenomena by a recent stochastic method. The results show that more approximate analyses for order parameters and correlation times can yield incorrect conclusions. As segmental motion decreases in rate, order parameters will be overestimated. Decreasing rates of pseudoisotropic motion lead to incorrect estimates of rotational correlation times. Either of the above can result in the inference of an artifactual phase transition in the lipids.     

Spectral Characterization of the Photoreducible b-Type Cytochrome of Dictyostelium discoideum

Irradiation of a soluble extract from broken cells of Dictyostelium discoideum causes the photoreduction of a b-type cytochrome. The cytochrome b can be separated from cytochrome c, which is also present in the extract, by column chromatography on Brushite, but the cytochrome b is no longer sensitive to light after separation on the column. Low temperature spectroscopy shows that reduced form of the photoreducible cytochrome b has a Soret band at 423 nm and a split α band with maxima at 558 and 551 nm similar to the b-type cytochrome in complex II of beef heart mitochondria.