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Biomedical subjects

M B Wall

Publications and source records attributed to M B Wall.

5 recordsLinked to original sources

Sensitivity to optic flow in human cortical areas MT and MST.

The macaque V5/MT complex comprises several sub-regions but little is known of their human homologues. We examined human V5/MT with fMRI in terms of specificity to optic flow stimuli, a key characteristic of macaque MST. Stimuli were large fields of moving dots, forming coherent global flow patterns. Random motion was used as a control. Retinotopic mapping was also conducted. The previously suggested existence of at least two distinct sub-regions, MT and MST, within the V5/MT complex was confirmed. Human MT is activated about equally by all moving dot patterns, including random motion, suggesting that it has little sensitivity to global flow structure. As previously described, this region shows strong signs of retinotopic organization and is only weakly activated by stimuli confined to the ipsilateral hemifield. In human MST, located immediately anterior to MT and strongly driven by ipsilateral stimuli, activation varies markedly with optic flow structure. The strongest activation is produced by complex flow that contains multiple flow components (expansion, contraction and rotation). Single components produce rather less response, while rigid translation and random motion produce less still. The results suggest that human MST is strongly specialized for encoding global flow properties, while human MT is less so.

Adult↗

DNA template amplification genetic studies on archival human preimplantation biopsied microtubed cell samples.

Biopsied cell samples can remain in storage following an initial, unequivocally successful preimplantation genetic diagnosis (PGD). The fidelity of the DNA template for polymerase chain reaction (PCR) amplification of microtubed human preimplantation embryonic cells stored at -70 degrees C for extended periods of time (6 months to 4 years) has been assessed. PCR protocols and specific nested primer sets for the beta-globin, ZP3 and CA repeat motif successfully used for previous human embryo PGD research were employed for these studies. The results show that the DNA template of microtubed biopsied blastomere and mural trophectoderm cell samples stored for periods of up to 4 years may be successfully amplified by PCR. Specific gene sequences were able to be analysed at the 1-2 or 3-5 biopsied cell level with a 71-100% success rate. Analysis of DNA fragments amplified from the CA dinucleotide repeat locus showed that in 8/9 samples both alleles were amplified at the cellular level. No DNA contamination was detected in the stored microtubed samples, or in the experimental controls.

Actins↗

Archival fixed and analysed preimplantation human embryonic cells: a DNA resource for retrospective PCR analysis at the cellular level.

Biopsies of human embryonic cell preparations previously analysed by cytogenetic and/or fluorescent in-situ hybridization (FISH) chromosome probes provide a unique reference DNA resource for the archival preimplantation genetic diagnosis (PGD) of the transferred embryo. DNA polymerase chain reaction (PCR) may be utilized on these fixed cell preparations to verify equivocal FISH/PGD results. Retrospective PCR screens of the genotype of biopsied embryonic cell(s) may be of benefit in the case of a suspected genetic mutation. Currently, carrier detection or linkage analysis is often not possible because of early death of the fetus, or of patients with a lethal disease. Alternatively, fixed/stained 'failed fertilized' oocytes provide a resource to extend genetic analysis to infertile patients. A successful research is described which minimizes loss of individual analysed fixed/stained oocytes, metaphase chromosomes, and embryonic cell samples. Initial DNA amplification takes place in situ using a modified PCR protocol. Comparative cellular studies using primer sets previously used for PGD analyses show that 65% of the preparations amplified unequivocally using the modified protocol and primers for a CA repeat motif gene sequence, in comparison with 81% using the original PCR protocol. With further refinement and optimization, the methods outlined have the potential to retrospectively screen archival fixed chromosomes, gametes, and embryonic cells for clinical application, and enable the further study of the fixed human preimplantation embryo at the morphological, cell and molecular level.

Biopsy↗

Cytogenetic and fluorescent in-situ hybridization chromosomal studies on in-vitro fertilized and intracytoplasmic sperm injected 'failed-fertilized' human oocytes.

This study was undertaken to establish baseline data on the chromosomal status of 'failed-fertilized' oocytes derived from in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) procedures. A cytogenetic analysis was undertaken on 162 IVF and 51 ICSI oocytes. In all, 82.1% (133/162) of the IVF and 78.4% (40/51) of the ICSI oocytes had metaphase II (MII) plates, of which 50.4% of the IVF and 47.5% of the ICSI oocytes were analysed further. Chromosomes of the G-group (21-22) were identified with the majority of the anomalies. No overall significant difference in the aneuploidy rate was found for the IVF (37.3%) of ICSI (31.6%) oocytes, or with maternal age. However, chromosome anomalies, e.g. diploidy, fragmented and broken chromatids, single sperm and oocyte chromatids, were found in oocytes from IVF patients aged > 36 years and in the ICSI oocytes throughout the maternal age range (31-38 years). The status of the polar body chromatin indicated that there was no overall significant difference in the maturation of the IVF and ICSI oocytes. Evidence of successful sperm delivery was found in 72.5% (37/51) of the ICSI failed-fertilized oocytes. In this group there was a significant increase in the incidence of premature chromosome condensation: 19.6% (10/51) contained sperm chromosomes, 7.8% (4/51) had swollen sperm heads, and the remaining 45.0% had condensed sperm heads. The presence of both sperm and MII oocyte chromosomes was found in 19.6% (10/51) of the ICSI and 8.6% (14/162) of the IVF failed-fertilized oocytes. Specific fluorescent in-situ hybridization DNA probes were used to re-analyse the chromosomes of karyotyped 'failed-fertilized' IVF oocytes and, for the first time, applied to the karyotyped chromosomes of failed-fertilized ICSI oocytes. The hybridization efficiency was 86-95% for the centromere probe and 100% for probes 21 and 18.

Adult↗

Biopulping process design and kinetics.

Biopulping is the solid-state fermentation of wood chips as a pretreatment for mechanical pulping processes. The two organisms that are currently of the greatest interest for biopulping are the white-rot fungi, Phanerochaete chrysosporium and Ceriporiopsis subvermispora. P. chrysosporium has been shown to successfully biopulp wood (33% energy savings; 39% improvement in tear index) without the need for sterilization of the wood or nutrient supplementation. Demonstrating the practical and economical feasibility of the biopulping process requires process modeling based on accurate kinetic data. Techniques to monitor dry weight loss and growth rate as functions of time using carbon dioxide production data have been developed. Growth was shown to be linear with time on unsupplemented chips and exponential with time on supplemented chips.

Journal Article↗