Isolation and partial characterization of host cell wall surface glycoproteins: their involvement in agglutination, attachment and appressorium formation by piptocephalis virginiana

Cell surface proteins obtained by alkaline extraction from isolated cell walls of Mortierella pusilla and M. candelabrum, host and nonhost, respectively, to the mycoparasite, Piptocephalis virginiana, were tested for their ability to agglutinate mycoparasite spores. The host cell wall protein extract had a high agglutinating activity (788 a.u. mg- t ) as compared with the nonhost extract (21 a.li. mg- t ). SDS-polyacrylamide gel electrophoresis of the cell wall proteins revealed four protein bands, a, b, c, and d (Mr 117, 100, 85 and 64 kd, respectively) at the host surface, but not at the nonhost surface, except for the faint band c. Deletion of proteins b or c from the host cell wall protein extract significantly reduced its agglutinating activity. Proteins band c, obtained as purified preparations by a series of procedures, were shown to be two glycoproteins. Carbohydrate analysis by gas chromatography demonstrated that glucose and Nacetylglucosamine were the major carbohydrate components of the glycoproteins. It was further shown that the agglutinating activity of the pure preparation containing both band c was 500-850 times that of the single glycoproteins, suggesting the involvement of both glycoproteins in agglutination. The results suggest that the glycoproteins band c are the two subunits of agglutinin present at the host cell surface. The two glycoproteins band c purified from the host cell wall protein extract were further examined after various treatments for their possible role in agglutination, attachment and appressorium formation by the mycoparasite. Results obtained by agglutination and attachment tests showed: (1) the two glycoprotein-s are not only an agglutinin responsible for the mycoparasite spore agglutination, but may also serve as a receptor for the specific recognition, attachment and appressorium formation by the mycoparasite; (2) treatment of the rnycoparasite spores with various sugars revealed that arabinose, glucose and N-acetylglucosamine inhibited the agglutination and attachment activity of the glycoproteins, however, the relative percentage of appressorium formation was not affected by the above sugars; (3) the two glycoproteins are relatively stable with respect to their agglutinin and receptor functions. The present results suggest that the agglutination and attachment may be mediated directly by certain sugars present at the host and mycoparasite cell surfaces while the appressorlum formation may be the response of complementary combinations of both sugar and protein, the two parts of the glycoproteins at the interacting surfaces of two fungi.

Vitamin D status and latent tuberculosis infection : a preliminary study in a group of healthy Mexican agricultural workers

Vitamin D metabolites are important in the regulation of bone and calcium homeostasis, but also have a more ubiquitous role in the regulation of cell differentiation and immune function. Severely low circulating 25-dihydroxyvitamin D [25(OH)D] concentrations have been associated with the onset of active tuberculosis (TB) in immigrant populations, although the association with latent TB infection (LTBI) has not received much attention. A previous study identified the prevalence of LTBI among a sample of Mexican migrant workers enrolled in Canada's Seasonal Agricultural Workers Program (SA WP) in the Niagara Region of Ontario. The aim of the present study was to determine the vitamin D status of the same sample, and identify if a relationship existed with LTBI. Studies of vitamin D deficiency and active TB are most commonly carried out among immigrant populations to non-endemic regions, in which reactivation of LTBI has occurred. Currently, there is limited knowledge of the association between vitamin D deficiency and LTBI. Entry into Canada ensured that these individuals did not have active TB, and L TBI status was established previously by an interferon-gamma release assay (IGRA) (QuantiFERON-TB Gold In-Tube®, Cellestis Ltd., Australia). Awareness of vitamin D status may enable individuals at risk of deficiency to improve their nutritional health, and those with LTBI to be aware of this risk factor for disease. Prevalence of vitamin D insufficiency among the Mexican migrant workers was determined from serum samples collected in the summer of 2007 as part of the cross sectional LTBI study. Samples were measured for concentrations of the main circulating vitamin D metabolite, 25(OH)D, with a widely used 1251 250HD RIA (DiaSorin Inc.®, Stillwater, MN), and were categorized as deficient «37.5 nmoI/L), insufficient (>37.5 nmollL, < 80 nmol/L) or sufficient (2::80 nmoI/L). Fisher's exact tests and t tests were used to determine if vitamin D status (sufficiency or insufficiency) or 25(OH)D concentrations significantly differed by sex or age categories. Predictors of vitamin D insufficiency and 25(OH)D concentrations were taken from questionnaires carried out during the previous study, and analyzed in the present study using multiple regression prediction models. Fisher's exact test and t test was used to determine if vitamin D status or 25(OH)D concentration differed by LTBI status. Strength of the relationship between interferongamma (IFN-y) concentration (released by peripheral T cells in response to TB antigens) and 25(OH)D concentration was analyzed using a Spearman correlation. Out of 87 participants included in the study (78% male; mean age 38 years), 14 were identified as LTBI positive but none had any signs or symptoms of TB reactivation. Only 30% of the participants were vitamin D sufficient, whereas 68% were insufficient and 2% were deficient. Significant independent predictors of lower 25(OH)D concentrations were sex, number of years enrolled in the SA WP and length of stay in Canada. No significant differences were found between 25(OH)D concentrations and LTBI status. There was a significant moderate correlation between IFN-y and 25(OH)D concentrations ofLTBI-positive individuals. The majority of participants presented with Vitamin D insufficiency but none were severely deficient, indicating that 25(OH)D concentrations do not decrease dramatically in populations who temporarily reside in Canada but go back to their countries of origin during the Canadian winter. This study did not find a statistical relationship between low levels of vitamin D and LTBI which suggests that in the presence of overall good health, lower than ideal levels of 2S(OH)D, may still be exerting a protective immunological effect against LTBI reactivation. The challenge remains to determine a critical 2S(OH)D concentration at which reactivation is more likely to occur.

Indenture between Samuel D. Woodruff and Terrance Johnson

Indenture of agreement for sale of land between Samuel D. Woodruff of St. Catharines and Terrance Johnson of St. Catharines for purchase of lot no. 20 in the 5th Concession in the Township of Grantham. This is accompanied by a printed letter which states the S.D. Woodruff is to make a conveyance of the lands to Edward McArdle. The letter is signed by T. Johnson, February 19, 1877.

Indenture between Samuel D. Woodruff and Hamilton Killaly Woodruff

Indenture of deed of land between Samuel DeVeaux of St. Catharines and Hamilton Killaly Woodruff of St. Catharines regarding parts of Lots numbered 20 and 21 in the 5th and 6th Concessions of the Township of Grantham and part of the road allowance between the aforementioned Concessions – instrument no. 9353, Jan. 17, 1894.

Indenture between Samuel D. Woodruff and Lewis Better

Indenture regarding articles of agreement between Samuel D. Woodruff of St. Catharines and Lewis Better of the Township of Gainsborough for lot no. 3 in the township of Gainsborough, Apr. 23, 1896.

Indenture between Samuel D. Woodruff and Jane Caroline Woodruff and Hamilton Killaly Woodruff

Indenture between Samuel D. Woodruff and Jane Caroline Woodruff and Hamilton Killaly Woodruff in which Samuel Woodruff signed over pew 15 in St. George’s Church, St. Catharines to Hamilton Killaly Woodruff for $1.00, May 12, 1900.

The letter is addressed to Samuel D. Woodruff and is signed by Thomas Steers

Letter which is very stained, fragile and has holes in it. Text is partially illegible and faded. The letter is addressed to Samuel D. Woodruff and is signed by Thomas Steers. It is written in ink, but someone has added comments in pencil. It regards Richard and William Woodruff. There are questions on the document which include: “Has M. Clement died interstate?” [in pencil – “he has”]; “Has he an heir at law other than” [the rest is faded, someone has written in pencil “he has, Richard and William Woodruff”], March 23, 1847.

Indenture of Agreement for Sale of Land between S.D. Woodruff and Elizabeth Cudney

Indenture of agreement for Sale of Land between S.D. Woodruff of St. Catharines and Elizabeth Cudney of Willoughby regarding a footpath and Lots 9 and 10 in Willoughby, Feb. 6, 1893.

Indenture of memorandum of an agreement between S.D. Woodruff and James L. Burton and M. Burton

Indenture of memorandum of an agreement between S.D. Woodruff of St. Catharines and James L. Burton and M. Burton, both of Barrie, trading under the name of Burton and Bro. that Burton and Bro. would buy all the pine timber located in berths 192 and 198. Burton and Bro. agrees to have all timber cut. The agreement is signed by S.D. Woodruff and Burton and Bro. This document is badly burned along the left hand side. This does not affect the text, July 11, 1877.

Agreement between S.D. Woodruff and Nathaniel Dyment of Barrie

Agreement between S.D. Woodruff and Nathaniel Dyment of Barrie. S.D. Woodruff is the owner of license no. 198 of season 1877/78 for berth no. 198. S.D. Woodruff has agreed to sell this license, Nov. 23, 1877.

Letter is addressed to S.D. Woodruff and is from Dunnville [signature illegible]

Letter is addressed to S.D. Woodruff and is from Dunnville [signature illegible] (3 pages, handwritten). The writer suggests that the head of the Welland Canal should be guarded. He has made a deal with the Sappers and Miners [Corps of Royal Military Artificers, consisting of noncommissioned officers and privates] to be stationed at this point, Jan.24, 1862.

List which includes costs, plans and receipts for amounts received between S.D. Woodruff and Boyd and Schurr

List (8 pages, handwritten) which includes costs, plans and receipts for amounts received between S.D. Woodruff and Boyd and Schurr, Dec. 3, 1875.

Flyer about railroad bonds from E.D. Shepard and Co. Bankers

Flyer (2 double sided pages, printed) about railroad bonds from E.D. Shepard and Co. Bankers, New York, n.d.

Hydration, conformational states and kinetics of yeast hexokinase PII /

The kinetic study of the coupled enzymatic reaction involving monomeric yeast hexokinase PII (HK) and yeast glucose-6-phosphate dehydrogenase (G-6-PDH) yields a Michaelis constant of 0.15 ± 0.01 mM for D-glucose. At pH 8.7 HK is present in monomeric form. The addition of polyethylene glycol (PEG), to the reaction mixture increased the affinity of HK for glucose, independent ofMW of the PEG from 2000 to 10000. The osmotic stress exerted by PEG can be used to measure the change in number of water molecules that accompany enzyme conformational changes (Rand, et al., 1993). Results indicate that the G-6-PDH is not osmotically sensitive and thus, the change in the number of PEG-inaccessible water molecules (ANw) measured in the coupled reaction is only the difference between the glucose-bound and glucosefree conformations of HK. ANw ~ 450 with PEGs of MW > 2000 under conditions for both binding (Reid and Rand, 1997) and kinetic assays. The contribution water may play in the binding of ATP (Km = 0.24 + 0.02 mM) has also been examined. It was found that in this case ANw = (for osmotic pressures < 2.8x10* dynes/cm^), suggesting no additional numbers of waters are displaced when ATP binds to HK. Osmotic pressure experiments were also performed with dimeric HK. It was determined that both the monomeric and dimeric forms of HK give the same ANw under low pressures. If this large ANw is due to conformational flexibility, it would appear that the flexibility is not reduced upon dimerization ofthe enzyme.

The influence of skeletal muscle cell volume on the regulation of carbohydrate uptake and muscle metabolism

This study investigated the regulation of carbohydrate metabolism and glucose uptake through changes in skeletal muscle cell volume. Using an established invitro isolated whole muscle model, soleus (SOL) and extensor digitorum longus (EDL) muscles were dissected from male rats and incubated in an organ bath containing Sigma medium-199 with 8 mM D-glucose altered to target osmolality (hypo-osmotic: HYPO, iso-osmotic: ISO, hyper-osmotic: HYPER; 190, 290, 400 mmol/kg). Muscles were divided into two groups; metabolite (MM) and uptake (MU). MM (N=48) were incubated for 60 minutes and were then immediately flash frozen. MU (N=24) were incubated for 30 minutes and then the extracellular fluid was exchanged for media containing ^H-glucose and ^'*C-mannitol and incubated for another 30 minutes. After the incubation, the muscles were freeze clamped. Results demonstrated a relative water decrease and increase in HYPER and HYPO, respectively. EDL and SOL glucose uptakes were found to be significantly greater in HYPER conditions. The HYPER condition resulted in significant alterations in muscle metabolite concentrations (lower glycogen, elevated lactate, and G-6-P) suggesting a catabolic cell state, and an increase in glycogen synthase transformation when compared to the HYPO group. In conclusion, skeletal muscle cell volume alters rates of glucose uptake with further alterations in muscle metabolites and glycogen synthase transformation.