Interactions of the DNA intercalator acridine orange, with itself, with caffeine, and with double stranded DNA

Caffeine (CAF) inhibits the intercalation of acridine orange (AO) into cellular DNA. Optical absorption and fluorescence spectroscopy were employed to determine the molecular interactions of AO with itself, with CAF, and with double stranded herring sperm DNA (dsDNA). AO dimerization was observed at concentrations >2 micromol. The sharp increase in fluorescence (lambda(em)=530 nm) at 5 micromol of AO was attributed to AO multimer formation. From 0.5 to 5.0 micromol, the AO self-association binding constant (K(assoc)) was determined to be 38620 mol(-1), however, the presence of 150 mmol NaCl increased K(assoc) to 118000 mol(-1) attributed to the charge neutralization. The K(assoc) for AO with CAF was confirmed to be 256 mol(-1). K(assoc) for the binding of AO with 20 micromol DNA ranged from, 32000 mol(-1) at 2 micromol AO, to approximately 3700 mol(-1) at 10 micromol AO, in the absence of NaCl. This AO concentration dependency of K(assoc) value with DNA was attributed to AO intercalation into dsDNA at high dsDNA/AO ratios, and electrostatic binding of AO to dsDNA at low AO ratios. The findings provide information used to explain fluorescence intensity values at lambda(em) at 530 nm from studies that combine AO, caffeine, and dsDNA.     

CLPB Variants Associated with Autosomal-Recessive Mitochondrial Disorder with Cataract,Neutropenia, Epilepsy,and Methylglutaconic Aciduria

3-methylglutaconic aciduria (3-MGA-uria) is a nonspecific finding associated with mitochondrial dysfunction, including defects of oxidative phosphorylation. 3-MGA-uria is classified into five groups, of which one, type IV, is genetically heterogeneous. Here we report five children with a form of type IV 3-MGA-uria characterized by cataracts, severe psychomotor regression during febrile episodes, epilepsy, neutropenia with frequent infections, and death in early childhood. Four of the individuals were of Greenlandic descent, and one was North American, of Northern European and Asian descent. Through a combination of homozygosity mapping in the Greenlandic individuals and exome sequencing in the North American, we identified biallelic variants in the caseinolytic peptidase B homolog (CLPB). The causative variants included one missense variant, c.803C>T (p.Thr268Met), and two nonsense variants, c.961A>T (p.Lys321^∗) and c.1249C>T (p.Arg417^∗). The level of CLPB protein was markedly decreased in fibroblasts and liver of affected individuals. CLPB is proposed to function as a mitochondrial chaperone involved in disaggregation of misfolded proteins, resulting from stress such as heat denaturation.     

Working with,not against,coral-reef fisheries

The fisheries policies of some Pacific island nations are more appropriate to the biology of their resources than are some of the fisheries policies of more industrialized countries. Exclusive local ownership of natural resources in Palau encourages adjustive management on biologically relevant scales of time and space and promotes responsibility by reducing the tragedy of the commons. The presence of large individuals in fish populations and adequate size of spawning aggregations are more efficient and meaningful cues for timely management than are surveys of abundance or biomass. Taking fish from populations more than halfway to their carrying capacity is working favorably with the fishery because removing fish potentially increases resource stability by negative feedback between stock size and population production. Taking the same amount of fish from a population below half its carrying capacity is working against the fishery, making the population unstable, because reducing the reproductive stock potentially accelerates reduction of the population production by positive feedback. Reef fish are consumed locally, while Palauan laws ban the export of reef resources. This is consistent with the high gross primary production with little excess net production from undisturbed coral-reef ecosystems. The relatively rapid growth rates, short life spans, reliable recruitment and wide-ranging movements of open-ocean fishes such as scombrids make them much more productive than coral-reef fishes. The greater fisheries yield per square kilometer in the open ocean multiplied by well over a thousand times the area of the exclusive economic zone than that of Palau’s coral reefs should encourage Palauans to keep reef fishes for subsistence and to feed tourists open-ocean fishes. Fisheries having only artisanal means should be encouraged to increase the yield and sustainability by moving away from coral reefs to bulk harvesting of nearshore pelagics.     

Kymographs,stimulators, and kymographs with stimulators

This biology investigation on Pristipomoides filamentosus larval development, survival, and aquaculture research was developed with three educational objectives: to provide high school students with (1) a scientific background on the biology and science of fisheries as well as overfishing, its consequences, and possible mitigations; (2) exposure to field and laboratory techniques in marine science; and (3) practical skills in scientific inquiry and investigation. We teach this investigation at the Hawai‘i Institute of Marine Biology, where we have access to captive broodstock of the pink snapper, P. filamentosus. During this investigation students follow several steps scientists take to study aquaculture: collecting spawn from outdoor fish pens, quantifying the number of eggs, determining the percentage of fertilisation, and estimating the time of spawning and hatching. Additionally, students perform hypothesis-driven science activities with the embryos to test the effects of water quality on their development and survival. In this paper we discuss background information of aquaculture, specifically of P. filamentosus, and thoroughly describe the several components of delivering the investigation. Lastly, we provide possible outcomes of students’ performances in the laboratory activities, and discuss how effectively the exercise met its educational objectives.     

Local vasodilatation with metacholine, but not with nitroprusside, increases forearm glucose uptake

Insulin is known to increase blood flow in parallel to glucose uptake in skeletal muscle. However, it is not known if an increase in blood flow by itself is associated with an increase in glucose uptake in the absence of hyperinsulinemia. To investigate further this matter, the effect of increased blood flow on forearm glucose uptake was studied in the fasting state during intra-arterial infusions of two different vasodilators, metacholine and nitroprusside, in 19 hypertensive subjects. Both metacholine (4 microg/min) and nitroprusside (10 microg/min) increased resting forearm blood flow, measured by venous occlusion plethysmography, to a similar degree (180 % and 170 %, respectively, p<0.0001 for both). However, metacholine infusion increased the forearm glucose uptake from 2.0+/-0.9 (S.D.) during rest to 5.5+/-3.0 umol/min/100 ml tissue (p<0.0001), while no significant change in glucose uptake was seen during nitroprusside infusion (2.3+/-1.4 micromol/min/100 ml tissue). In conclusion, vasodilatation induced by metacholine, but not by nitroprusside, increased glucose uptake in the forearm of hypertensive patients. Thus, an increase in forearm blood flow does not necessarily improve glucose uptake in the forearm during the fasting state.     

Studies with a plant fungicide,imazalil, with vapor-phase activity,in the therapy of human alternariosis

The use of a plant fungicide, the imidazole imazalil, in the therapy of human alternariosis is described. The patient's disease involved the palate, nose and sinuses, and had been unresponsive to conventional therapy. Imazalil has been used to treat Alternaria infections of plant materials. The patient's isolate was susceptible in vitro. We also showed that the drug was inhibitory to fungi in vitro as a vapor, not requiring contact by diffusion through agar or broth. The drug was used topically, by installation and irrigation, in this case and administered orally in doses of up to 1 200 mg daily, for 6 months. Peak serum concentrations exceeded 4 meg/ml. The infection was arrested but not cured. The drug was tolerated without evident toxicity, though an unpleasant taste and nausea at high doses were limiting.     

Obesity profiles with knee osteoarthritis: correlation with pain, disability, disease progression

Objectives: To identify the prevalence of overweight among community‐dwelling adults diagnosed as having knee osteoarthritis (OA) and the relationship between the weight status of these individuals, selected disease‐related outcomes, and disease progression. Research Methods and Procedures: The BMIs of 82 women and 18 men with unilateral or bilateral knee OA were examined on a single occasion along with data on physical comorbidities, pain, and function and subjected to correlation analyses. BMIs from two additional samples, one that included 16 women with and without knee OA and one that included 24 women and 6 men with knee joint OA that required surgery for the subsequent onset of hip OA, were also assessed. Results: At least 80% of all present cohorts were overweight or obese. Those with higher BMIs reported more pain than those with lower BMIs (p < 0.05) and pain was related to perceived physical exertion (p < 0.05). Body mass indices were not significantly correlated with generic gait measures, but an inverse trend toward the time spent in the gait cycle (r = ?0.63; p = 0.097) that may impact the disease process was identified. Those with comorbidities had the same body mass, on average, as those with no comorbidities, and those with bilateral disease were heavier than those with unilateral disease. Discussion: A high body mass is present in most adults with knee OA. Moreover, being overweight may affect knee joint impact rates and pain incrementally. Having high body weights may heighten the risk for bilateral knee joint, as well as hip joint, OA.