Creation of Chitinase Producer and Disruption of Micromycete Cell Wall with the Obtained Enzyme Preparation.

A recombinant strain producing a complex of extracellular enzymes including chitinase from Myceliophtora thermophila was created based on the fungus Penicillium verruculosum. The activity of the enzyme preparations obtained from the cultural fluid of the producer strain was 0.55, 0.53, and 0.66 U/mg protein with chitin and chitosans with the molecular weight of 200 and 1000 kDa, respectively. The temperature optimum for the recombinant chitinase was 52-65°C; the pH optimum was 4.5-6.2, which corresponded to the published data for this class of the enzymes. The content of heterologous chitinase in the obtained enzyme preparations was 47% of total protein content in the cultural fluid. Enzyme preparations produced by the recombinant P. verruculosum XT403 strain and containing heterologous chitinase were able to degrade the mycelium of micromycetes, including phytopathogenic ones, and were very efficient in the bioconversion of microbiological industry waste.

In vivo burn diagnosis by camera-phone diffuse reflectance laser speckle detection.

Burn diagnosis using laser speckle light typically employs widefield illumination of the burn region to produce two-dimensional speckle patterns from light backscattered from the entire irradiated tissue volume. Analysis of speckle contrast in these time-integrated patterns can then provide information on burn severity. Here, by contrast, we use point illumination to generate diffuse reflectance laser speckle patterns of the burn. By examining spatiotemporal fluctuations in these time-integrated patterns along the radial direction from the incident point beam, we show the ability to distinguish partial-thickness burns in a porcine model in vivo within the first 24 hours post-burn. Furthermore, our findings suggest that time-integrated diffuse reflectance laser speckle can be useful for monitoring burn healing over time post-burn. Unlike conventional diffuse reflectance laser speckle detection systems that utilize scientific or industrial-grade cameras, our system is designed with a camera-phone, demonstrating the potential for burn diagnosis with a simple imager.

H19 expression in hepatic metastases from a range of human carcinomas.

AIMS: To investigate the expression of the imprinted oncofetal H19 gene in hepatic metastases derived from a range of human carcinomas and assess its prognostic value with the view of developing a DNA based treatment for such metastases. METHODS: Non-radioactive in situ hybridisation for H19 RNA was performed on paraffin wax embedded sections of liver biopsies or partial hepatectomy specimens, taken from 80 patients with hepatic metastases derived from carcinomas from several medical centres in Israel. The degree of expression was graded qualitatively according to the number of cells expressing H19 and the intensity of staining. The medical files were searched for demographic data and survival times before and after diagnosis of hepatic metastases. RESULTS: H19 expression was found in the hepatic metastases of 64 of 80 patients. High expression (higher staining grades) of H19 in the metastases was found in 43 of 80 patients. However, H19 expression status in the hepatic metastases did not correlate with either the length of time to development of metastasis or overall survival. CONCLUSIONS: H19 is highly expressed in more than half of hepatic metastases derived from a range of carcinomas. Thus, these metastases may be suitable candidates for H19 DNA based treatment. Further studies are needed to determine whether H19 expression has prognostic value in metastatic liver disease using larger numbers of specific subtypes of primary carcinomas.

FT-IR microscopic characterization of normal and malignant human colonic tissues.

Fourier-transform infrared spectroscopy (FT-IR) employs a unique approach to optical diagnosis of tissue pathology based on the characteristic molecular vibrational spectra of the tissue. In this study, we report infrared absorption spectra of formalin fixed, paraffin embedded normal and malignant human colonic tissues from ten different patients. Our method is based on microscopic infrared study (FT-IR-microscopy) of thin tissue specimens in parallel with normal histopathological analysis, which serves as a reference. Our results indicate that the normal colonic tissue has a stronger absorption than the cancerous type over a wide region in all ten cases. The detailed analysis showed that there is a significant decrease in carbohydrate levels, total phosphate and also possibly creatine contents for cancerous tissue in comparison to the controls. Also, RNA/DNA ratio increased in cancerous tissues relative to the normals in all the patients. The results of Linear Discriminant Analysis (LDA) showed that the normal and malignant cells could be identified with about 89% accuracy.