Human UC-blood banking: impact of blood volume, cell separation and cryopreservation on leukocyte and CD34(+) cell recovery.

BACKGROUND: UC blood represents an increasingly useful source of hematopoietic stem cells for BMT, although, currently, low cell numbers generally limit its use to pediatric patients. We have determined parameters that influence the recovery of viable cells during processing and cryopreservation, in an effort to set guidelines for determining whether a sample will yield sufficient cells to be of use in the transplant setting. METHODS: UC blood was collected from donors from January 1996 to December 1999. Volume was reduced using Ficoll, followed by cryopreservation under liquid nitrogen. Total leukocyte and CD34(+)-cell counts were determined prior to processing and a subset of samples was also assessed post-processing and post-cryopreservation. RESULTS: Outcomes for 3816 samples were analyzed to determine the correlation between cell number, cell type, volume, and time between collection and processing. A positive relationship was observed between volume and cell number for both leukocytes and the CD34(+) cells. This correlation allowed us to determine the number of leukocytes and CD34(+) cells expected from a sample based on volume, and to set guidelines for determining the practicality of storing any given sample prior to processing and cryopreservation. DISCUSSION: Measuring blood volume gives a very useful indication of the total leukocyte and CD34(+) cell number. The majority (75%) of cord-blood samples contain sufficient leukocytes for a pediatric transplant, and the number of cells available can be determined prior to processing by measuring blood volume.

Regeneration of vas deferens after vasectomy.

A total of 338 specimens obtained from 182 patients undergoing reversal of previous vasectomies have been studied. The major histologic features are sprouting of tubules and extravasation of sperm. Neo-tubules grow only from the proximal end of the divided vas and were seen in 136 specimens. Sperm were present in the neo-tubules in 61 specimens, but in the lumen of the proximal vas in 88 per cent of all patients. Extravasation of sperm had occurred from the neo-tubules in 64 specimens. In 57/154 patients operated on bilaterally at the same time, regeneration was unilateral. The conclusion from the analysis of factors responsible for this selectively proximal process is that sperm secretion was probably a major factor but was certainly not the only factor responsible.

Pathologic conditions of the pisiform and pisotriquetral joint.

In eight of sixteen patients with symptomatic pisotriquetral joints the pisiform was excised. A detailed study allowed correlation of the etiologic factors with the pathologic diagnosis. In a similar fashion pathologic-etiologic data were retrieved from 216 cases identified from the world literature and organized into the following four pathologic groups: primary OA (2.3%), secondary OA (48.4%), other arthritides (4.7%), and flexor carpi ulnaris enthesopathy (44.6%). The most common causes were acute and chronic trauma and instability. On the basis of these data, we hypothesized that loss of integrity to the surrounding retinacular structures of the pisiform may lead to instability and thus dysfunction of the joint.

Centrode patterns and segmental instability in degenerative disc disease.

Moiré fringes were used to determine centrode patterns in cadaveric spines with degenerative disc disease. The normal centrodes were compared with those with minor, mild, moderate, and severe degenerative disc disease. The most complex loci noted were those discs that showed minimal radiographic evidence of degeneration, that is, the minor and mild groups. The loci lengthened significantly (P less than 0.001) when compared with normal controls. The position of the centrode shifted downward into the body of L5 in the moderate group.

Mineral oil (follicular) lipidosis: II. Histologic studies of spleen, liver, lymph nodes, and bone marrow.

Tissues obtained from 600 routine autopsies were studied. Mineral oil lipidosis was present in the spleen (76 per cent), liver (45 per cent), bone marrow (26 per cent), and lymph nodes; more than 50 per cent of the lymph nodes from the mesentery, porta hepatis, and mediastinum were affected. Mineral oil and its metabolic products produce a nonfibrogenic reaction in these tissues. The differential diagnosis of mineral oil lipidosis in lymph nodes with reaction to radiopaque oils and Whipple's disease is discussed. The presence of mineral oil in para-aortic (42 per cent) and internal iliac (15 per cent) lymph nodes could result in false-positive readings after lymphangiography.

Follicular (mineral oil) lipidosis: I. Epidemiologic studies of involvement of the spleen.

A histologic study of spleens from autopsies performed in 74 centers in 41 countries showed the incidence of follicular (mineral oil) lipidosis to vary from 50 per cent (North America, Australia, New Zealand) to nil (Africa, Central and South America) in 1970 and 1971. Analysis of the factors responsible for the condition leads to the conclusion that the lipid is ingested from sources related to the packaging and display of foodstuffs.

Nasolacrimal bone changes in diseases of the lacrimal drainage system.

The bone pieces removed from 71 patients who had undergone dacryocystorhinostomies (DCR) or canaliculodacryocystorhinostomies (CDCR) were examined pathologically. Nineteen percent of cases showed evidence of active bone remodeling, 23% had immature, woven bone present, and the remaining cases showed normal mature cortical and cancellous bone. There were no inflammatory changes found. The clinical histories of the patients were reviewed and related to the bone findings. The clinical applications of the bone changes are discussed.

Determination of a locus of instantaneous centers of rotation of the lumbar disc by moiré fringes. A new technique.

The aim of this study was to define the motion of the L4-5 spinal segment moving from full extension to full flexion. A new technique using moiré fringes was applied to measure angles of 3-degree change, a technique proven to be highly accurate for small angular changes. This investigation has identified a locus of centers of rotation (centrode) that is located in the posterior half of the intervertebral disc. The direction of the locus passes from posterior to anterior and back to posterior. The technique can be adapted for evaluation of in vivo radiographs. This technique may prove useful in identifying aberrations in movement associated with spinal segment instability in its earliest stages.

Local modulation of progesterone production in human fetal membranes.

We have studied the ability of human fetal membranes (amnion, chorion, and decidua) to produce progesterone. Membranes were obtained from women at 38-41 weeks pregnancy following spontaneous onset of labor. The tissues were dispersed into single cell preparations using 0.05% collagenase in a Krebs-Ringer buffer system. Progesterone was measured by specific RIA. All 3 tissues produced progesterone in a significant dose-response relationship with added pregnenolone but chorion was significantly more active than the other membranes. Equimolar concentrations of dehydroepiandrosterone, estrone or estradiol cause significant inhibition (to 50%) of this activity but dehydroepiandrosterone sulfate and estriol did not. We conclude that human fetal membranes at term can produce progesterone and this production may be modulated by other steroids of fetal or maternal origin. Local regulation of progesterone production, without significant changes in circulating levels of progesterone, may have an important influence on local prostaglandin generation and hence on myometrial contractility.

A clinical and experimental investigation of occult injuries of the pelvic ring.

Apparently isolated fractures of the pubic rami must be associated with damage elsewhere in the pelvic ring. In a clinical review of 20 patients it was demonstrated that radiographic evidence of isolated fractures of the pubic rami was associated with the increased uptake of technetium polyphosphate on bone scans in either the sacroiliac joints or the acetabulum. The nature of the injury causing increased uptake was evaluated by means of an experimental study creating pelvic injuries in rabbits. Radiographs, bone scans, autoradiographs and histological sections of the pelves of these animals all showed evidence of microavulsion fractures of the subchondral bone mediated through the sacroiliac ligaments and Sharpey's fibres. The associated injuries account for the clinical picture in patients who complain of pain in the hip and sacroiliac region following fractures of the pubic rami. The clinical and radiographic evidence presented clearly demonstrates that an injury at one site in the pelvic ring must be associated with another on the other side of the ring. Furthermore, the technique developed for autoradiography of the animal pelves provides a new and useful tool for further study of injury to bones and ligaments.

Altered lymphocyte response in adjuvant-induced arthritis.

Evaluation of lymphocyte response has been undertaken in adjuvant-induced arthritis in rats. Lymphocyte reactivity was studied by evaluating lymphocyte transformation in mixed lymphocyte-chondrocyte cultures as well as the response to PHA and M. tuberculosis. It was found that animals immunized with Freund's complete adjuvant showed increased transformation when challenged with M. tuberculosis powder. However, there was a depressed response to PHA and a markedly depressed response in mixed lymphocyte-chondrocyte cultures.

The response of bone apposition rate to some nonphysiologic conditions.

The response of bone apposition to some nonphysiologic conditions was investigated. In rabbits, the normal osteogenic rhythm was totally abolished shortly following treatment of hydrocortisone at a dose of 2 mg/kg body weight daily. A new rhythm developed after the treatment was continued for 20 1/2--34 days. The rate of bone apposition was significantly depressed compared to normal data reported previously. In rats receiving one U.S.P. unit of parathyroid extract daily for 14 days, the rate of bone apposition was significantly higher than that in control rats. In both sepcies, the response to an exogenous hormone was identified for all skeletal locations. It was concluded that new bone apposition was a function of activated osteoblasts, and that this function responded only to general body control mechanisms.

Bone apposition rate as an index of bone metabolism.

The rate of bone apposition was studied by labeling bone with four or five sequential doses of oxytetracycline given at equal intervals of 2 or 4 days in rats and rabbits. The rate was estimated in bone formation sites bearing all the doses. This method has the advantage of assessing the rate of new bone mineralization during the active bone-forming phase only. This index of bone formation was found to be independent of the location of bone samples in the skeleton, and therefore might be under the influence of general body mechanisms only. Preliminary study in the rats indeed yielded results indicating that this bone apposition rate is accelerated by dietary deficiency in calcium.