The white-rot fungus, Phanerochaete chrysosporium, under combinatorial stress produces variable oil profiles following analysis of secondary metabolites.

AIMS: The effects of combinatorial stress on lipid production in Phanerochaete chrysosporium remain understudied. This species of white-rot fungi was cultivated on solid-state media while under variable levels of known abiotic and biotic stressors to establish the effect upon fungal oil profiles. METHODS AND RESULTS: Environmental stressors induced upon the fungus included the following: temperature, nutrient limitation and interspecies competition to assess impact upon oil profiles. Fatty acid type and its concentration were determined using analytical methods of gas chromatography and mass spectrometry. Growth rate under stress was established using high-performance liquid chromatography with ergosterol as the biomarker. Fungi grown on solid-state agar were able to simultaneously produce short- and long-chain fatty acids which appeared to be influenced by nutritional composition as well as temperature. Addition of nitrogen supplements increased the growth rate, but lipid dynamics remained unchanged. Introducing competition-induced stress had significantly altered the production of certain fatty acids beyond that of the monoculture while under nutrient-limiting conditions. Linoleic acid concentrations, for example, increased from an average of 885 ng µl-1 at monoculture towards 13 820 ng µl-1 at co-culture, following 7 days of incubation. CONCLUSIONS: Interspecies competition produced the most notable impact on lipid production for solid-state media cultivated fungi while the addition of nitrogen supplementation presented growth and lipid accumulation to be uncorrelated. Combinatorial stress therefore influences the yield of overall lipid production as well as the number of intermediate fatty acids produced, deriving similar oil profiles to the composition of vegetable and fish oils. SIGNIFICANCE AND IMPACT OF THE STUDY: Fungal secondary metabolism remains highly sensitive following combinatorial stress. The outcome impacts the research towards optimizing fungal oil profiles for biomass and nutrition. Future investigations on fungal stress tolerance mechanisms need to address these environmental factors throughout the experimental design.

The importance of mRNA structure in determining the pathogenicity of synonymous and non-synonymous mutations in haemophilia.

INTRODUCTION: Mutational analysis is commonly used to support the diagnosis and management of haemophilia. This has allowed for the generation of large mutation databases which provide unparalleled insight into genotype-phenotype relationships. Haemophilia is associated with inversions, deletions, insertions, nonsense and missense mutations. Both synonymous and non-synonymous mutations influence the base pairing of messenger RNA (mRNA), which can alter mRNA structure, cellular half-life and ribosome processivity/elongation. However, the role of mRNA structure in determining the pathogenicity of point mutations in haemophilia has not been evaluated. AIM: To evaluate mRNA thermodynamic stability and associated RNA prediction software as a means to distinguish between neutral and disease-associated mutations in haemophilia. METHODS: Five mRNA structure prediction software programs were used to assess the thermodynamic stability of mRNA fragments carrying neutral vs. disease-associated and synonymous vs. non-synonymous point mutations in F8, F9 and a third X-linked gene, DMD (dystrophin). RESULTS: In F8 and DMD, disease-associated mutations tend to occur in more structurally stable mRNA regions, represented by lower MFE (minimum free energy) levels. In comparing multiple software packages for mRNA structure prediction, a 101-151 nucleotide fragment length appears to be a feasible range for structuring future studies. CONCLUSION: mRNA thermodynamic stability is one predictive characteristic, which when combined with other RNA and protein features, may offer significant insight when screening sequencing data for novel disease-associated mutations. Our results also suggest potential utility in evaluating the mRNA thermodynamic stability profile of a gene when determining the viability of interchanging codons for biological and therapeutic applications.

Towards waterjet dissection in neurosurgery: experimental in-vivo results with two different nozzle types.

Waterjet dissection is under close investigation as a new neurosurgical tool. Experimentally, a precise tissue dissection with vessel preservation has been demonstrated in the porcine cadaver brain. The safety of the device has been shown in first clinical applications. However, a detailed in-vivo analysis of the waterjet device is still awaited. In the present study, two often applied nozzle types (100 microm diameter emitting a coherent straight jet; 120 microm diameter emitting a helically rotating jet) were experimentally studied in vivo. Forty-one rabbits received a frontal waterjet corticotomy on either side with one nozzle type after microsurgical removal of the arachnoid membranes. Animals were sacrificed at 1, 3, 7 days and 6 weeks after surgery. Dissection morphology and vessel preservation were evaluated. Tissue trauma was analyzed by the extent of intra-operative haemorrhage, postsurgical oedema formation and astrocytic as well as microglial reactions. In all animals, reliable brain dissection was observed. Macroscopically, only minor bleeding occurred. Microscopically, also very precise brain dissection with both nozzle types was found. Vessels were preserved with both pressures applied (5 and 10 bar). Dissections with the 100 microm straight nozzle were more precise with respect to dissection margins. However, no significant difference in vessel preservation and extent of haemorrhage, oedema formation, astrocytic and microglial reactivity was shown. Malfunction defined as clotting of the instrument occurred only with the 100 microm nozzle. In four 100 microm straight nozzle hemispheres, even no brain dissection was seen. The results indicate that the waterjet enables very precise and reliable brain parenchyma dissection with minimal trauma and vessel preservation in vivo. If this can be proven to be of clinical relevance, the instrument will become a valuable neurosurgical tool. Based on these results, the authors selected the 120 microm Helix nozzle for further research with this device in the CNS.

The diagnosis and treatment of central giant cell granuloma.

This report reviews the literature involving the central giant cell granuloma. Diagnosis and treatment is presented. The article reports the case of a child who was initially seen in her general dentist's office, then referred to an oral and maxillofacial surgeon. Differential diagnoses of both benign and malignant lesions related to the central giant cell tumor are discussed.

Quantitative ingrowth analysis of a porous-coated acetabular component in a canine model.

Histomorphometric analysis of bone ingrowth into a porous-coated acetabular component was evaluated in a canine model. A total of nine prostheses were evaluated, 3 at 6 months, 3 at 12 months, and 3 at 24 months after implantation. All implants were grossly stable at the time of retrieval. The mean percentage of bone ingrowth was 12% at 6 months, 24% at 12 months, and 24% at 24 months. Narrow radiolucent lines noted on microradiographs were more evident at the 6 month time period than at the 12 or 24 month time periods. Bone ingrowth into a porous-coated acetabular component in a weight-bearing model may continue beyond the 6 month time period, and early evaluation of bone ingrowth may underestimate final bone ingrowth.

In vitro loss of tensile strength and elasticity of five absorbable suture materials in sterile and infected canine urine.

The loss of breaking strength and elasticity of five absorbable suture materials (polydioxanone [PDS-II], polyglycolic acid [PGA], polyglactin 910 [PG-910], polyglyconate [GTMC], and chromic gut) after in vitro incubation in sterile, Escherichia coli- and Proteus mirabilis-inoculated canine urine was studied. Biomechanical testing, in a controlled environment, was performed during the 28-day study period. Polydioxanone and chromic gut retained greater than 90% of their original strengths after 28 days of incubation in sterile urine and 87% of original strengths in E. coli-inoculated urine. Polyglyconate retained 24% and 18% of original strength, respectively, after incubation in sterile and E. coli-inoculated urine for 28 days. Polyglycolic acid and PG-910 retained less than 30% of original strength in sterile urine and only 7% in E. coli-inoculated urine after 21 days of incubation. In P. mirabilis-inoculated urine, loss of tensile strength and elongation was significant for all suture materials. Polyglycolic acid and PG-910 lost all strength after 24 hours of incubation. Polydioxanone lost all strength after 7 days of incubation, whereas GTMC retained 19% at day 7. Chromic gut retained 78% at day 7 and 16% after 21 days of incubation, however, the absence of normal phagocytic destruction of chromic gut in this in vitro study may have artificially elevated these values. In sterile urine with chemically modified pH, loss of strength and elongation was greater in alkaline urine than in neutral or acidic urine for all types of suture materials.

Hypokalemic periodic paralysis: two case reports.

We report two cases of hypokalemic periodic paralysis in children presenting to the emergency department with diffuse weakness and no antecedent history of neurologic disease. The literature is briefly reviewed. Any child with acute weakness and normal mental status should undergo serum potassium determination to allow prompt diagnosis and therapy.