Radiopaque alginate microcapsules for X-ray visualization and immunoprotection of cellular therapeutics.

Alginate-poly-L-lysine-alginate (APA) microcapsules have been explored as vehicles for therapeutic drug and cell delivery. The permselectivity of these capsules provides a unique means of controlled drug release and immunoisolation of encapsulated cells. Immunoisolation is especially attractive as it abrogates the need for chronic immunosuppressive therapy and opens up the possibility for the delivery of numerous cell sources including xenogeneic grafts. APA microcapsules containing cellular therapeutics have proven effective in the short-term treatment of a wide range of diseases requiring enzyme or endocrine replacement therapy, including type I diabetes. If these microcapsules could be noninvasively monitored with X-ray imaging modalities (i.e., fluoroscopy, CT, and digital subtraction angiography), questions such as the ideal transplantation site, the best means of delivery, and the long-term survival of grafts could be better addressed. We have developed two novel alginate-based radiopaque microcapsule formulations containing either barium sulfate (Ba X-Caps) or bismuth sulfate (Bi X-Caps). As compared to conventional, nonradiopaque APA capsules, Ba X-Caps and Bi X-Caps containing human cadaveric islets resulted in a decrease in cellular viability of less than 5% up to 14 days after encapsulation. Both radiopaque capsules were found to be permeable to lectins < or =75 kDa, but were impermeable to lectins > or =120 kDa, thus ensuring the blockage of the penetration of antibodies while allowing free diffusion of insulin and nutrients. The glucose-responsive insulin secretion of the radiopaque encapsulated human islets was found to be unaltered compared to that of unlabeled controls, with human C-peptide levels ranging from 3.21 to 2.87 (Ba X-Caps) and 3.23 to 2.87 (Bi X-Caps) ng/islet at 7 and 14 days postencapsulation, respectively. Using fluoroscopy, both Ba X-Caps and Bi X-Caps could be readily visualized as single radiopaque entities in vitro. Furthermore, following transplantation in vivo in mice and rabbits, single capsules could be identified with no significant change in contrast for at least 2 weeks. This study represents the first attempt at making radiopaque microcapsules for X-ray guided delivery and imaging of cellular therapeutics. While human cadaveric islets were used as a proof-of-principle, these radiopaque capsules may have wide ranging therapeutic applications for a variety of cell types.

Effect of milking frequency on DHI performance measures.

Increasing production by increasing milking frequency (MF) is a management option available to dairy producers. This study examined effects of MF and interactions with region and herd size on measures of herd performance. Dairy Herd Improvement (DHI) Holstein herd summary records (n = 10,754, 10,550, and 10,438) for the years 1998, 1999, and 2000 were classified by MF: two times a day (2X) milking vs three times a day (3X); herd size: small (< 250 cows) and large (> or = 250 cows); and region: North and South. Percentage of herds milking 3X by year were 7.0, 6.7, and 7.1. Rolling herd average milk production was 16, 16, and 15% higher for herds milking 3X than herds milking 2X for the respective years. Herds milking 3X in the North region outproduced herds milking 3X in the South region. Milk fat and protein percentages were lower for herds milking 3X during all 3 yr. Differences in energy-corrected milk production between herds milking 3X and herds milking 2X were 14.5, 13.4, and 13.4% during the respective 3 yr as a result of lower component percentages for herds milking 3X. Herds milking 3X had more days open and higher actual calving intervals than herds milking 2X. Services per pregnancy for herds breeding primarily by artificial insemination were higher for herds milking 3X than for herds milking 2X. Somatic cell scores and weighted somatic cell counts were lower for herds milking 3X than herds milking 2X. Herds milking 3X had a higher percentage of somatic cell scores in the low range (0 to 3) and a lower percentage in the high range (7 to 9). Mean percentages of cows entering and leaving the herd were higher for herds milking 3X during all 3 yr.

Serum progesterone and milk production and composition in dairy cows fed two concentrations of nitrate.

Forty clinically normal lactating Holstein cattle from a herd involved in a natural outbreak of chronic nitrate toxicosis were divided into 2 equal groups according to production, stage of lactation, age, and apparent pregnancy state (pregnant or nonpregnant). One group was fed a low-nitrate ration (average of 356 ppm on dry matter basis in concentrate; less than 400 ppm in free-choice hay for 1st 5 wks of study). The 2nd group was fed a high-nitrate (HN) ration (average of 1,600 ppm in protein concentrate-amemded corn silage; 4,000 ppm in free-choice hay for the 8-week study). At the end of the study, the 2 groups were classified according to their starting reproductive status: nonpregnant (open); early pregnant (less than 60 da); midpregnant (average of 105 da). Milk production, milk fat, and milk nitrate concentrations were similar for cows fed both rations. Serum progesterone concentration (SPC) was depressed (P less than 0.05) in cows fed the HN ration. This effect was prominent in open, luteal phase cows, less prominent but still apparent in early pregnant cows, and absent in midpregnant cows. The early reproductive problems of chronic nitrate toxicosis may be due to depression of SPC. A possible mechanism of inhibition of luteal progesterone synthesis by inhibition of cytochrome P-450 is presented.