Physiology and cryosensitivity of coral endosymbiotic algae (Symbiodinium).

Coral throughout the world are under threat. To save coral via cryopreservation methods, the Symbiodinium algae that live within many coral cells must also be considered. Coral juvenile must often take up these important cells from their surrounding water and when adult coral bleach, they lose their endosymbiotic algae and will die if they are not regained. The focus of this paper was to understand some of the cryo-physiology of the endosymbiotic algae, Symbiodinium, living within three species of Hawaiian coral, Fungia scutaria, Porites compressa and Pocillopora damicornis in Kaneohe Bay, Hawaii. Although cryopreservation of algae is common, the successful cryopreservation of these important coral endosymbionts is not common, and these species are often maintained in live serial cultures within stock centers worldwide. Freshly-extracted Symbiodinium were exposed to cryobiologically appropriate physiological stresses and their viability assessed with a Pulse Amplitude Fluorometer. Stresses included sensitivity to chilling temperatures, osmotic stress, and toxic effects of various concentrations and types of cryoprotectants (i.e., dimethyl sulfoxide, propylene glycol, glycerol and methanol). To determine the water and cryoprotectant permeabilities of Symbiodinium, uptake of radio-labeled glycerol and heavy water (D(2)O) were measured. The three different Symbiodinium subtypes studied demonstrated remarkable similarities in their morphology, sensitivity to cryoprotectants and permeability characteristics; however, they differed greatly in their sensitivity to hypo- and hyposmotic challenges and sensitivity to chilling, suggesting that standard slow freezing cryopreservation may not work well for all Symbiodinium. An appendix describes our H(2)O:D(2)O water exchange experiments and compares the diffusionally determined permeability with the two parameter model osmotic permeability.

Phenotypic and population differences in the association between CILP and lumbar disc disease.

BACKGROUND: Lumbar disc disease (LDD) is one of the leading causes of disability in the working-age population. A functional single-nucleotide polymorphism (SNP), +1184T-->C, in exon 8 of the cartilage intermediate layer protein gene (CILP) was recently identified as a risk factor for LDD in the Japanese population (odds ratio (OR) 1.61, 95% CI 1.31 to 1.98), with implications for impaired transforming growth factorbeta1 signalling. AIM: To validate this finding in two different ethnic cohorts with LDD. METHODS: This SNP and flanking SNPs were analysed in 243 Finnish patients with symptoms of LDD and 259 controls, and in 348 Chinese subjects with MRI-defined LDD and 343 controls. RESULTS AND CONCLUSION: The results showed no evidence of association in the Finnish (OR = 1.35, 95% CI 0.97 to 1.87; p = 0.14) or the Chinese (OR = 1.05, 95% CI 0.77 to 1.43; p = 0.71) samples, suggesting that cartilage intermediate layer protein gene is not a major risk factor for symptoms of LDD in Caucasians or in the general population that included individuals with or without symptoms.

Optimal conditions for detection of reverse transcriptase activity in human placentas.

Optimal conditions for detecting reverse transcriptase activity in human placental extracts are described. They vary with the state of the placenta at birth and are influenced by relative amounts of detergent, monovalent cation, and protein in the reaction mixture. Demonstrating activity of the placental enzyme requires detergent, but the enzyme is sensitive to high detergent concentrations. This sensitivity can be be altered by lowering the monovalent cation concentration from 0.154 to 0.034 M and by adding protein to the reaction mixture. The detection of reverse transcriptase in Rauscher murine leukemia virus and baboon endogenous type C virus, but not in the Mason-Pfizer monkey type D virus, shows similar requirements.

Stage IA to IIB mediastinal Hodgkin's disease: three-dimensional volumetric assessment of response to treatment.

From 1979 to 1986, the response to treatment of 53 patients with stage IA to IIB mediastinal Hodgkin's disease was evaluated by three-dimensional volumetric analysis using thoracic computed tomographic (CT) scans. The mean initial volume of mediastinal disease in 34 patients treated with mantle and para-aortic irradiation was 166 mL, whereas for 19 patients treated with two to six cycles of multiagent chemotherapy and mantle and para-aortic irradiation the mean initial volume was 446 mL. Preliminary data suggested that patients with mediastinal volumes of less than 200 mL had a lower mediastinal relapse rate (13%) than patients with volumes greater than 200 mL (32%). For 12 patients receiving six cycles of nitrogen mustard, vincristine, procarbazine, and prednisone (MOPP), those with a greater than 85% reduction in volume 1 to 2 months after chemotherapy had a lower incidence of mediastinal relapse (zero of six, 0%) compared with patients having 85% or less reduction in volume (four of six, 67%). The primary value of this technique is that it provides a sensitive assessment of response to treatment and may aid in monitoring the effectiveness of a given treatment.

Biomechanical and electromyographic evaluation of ankle foot orthosis and dynamic ankle foot orthosis in spastic cerebral palsy.

This study evaluated the biomechanical and electromyographic effects of conventional ankle foot orthoses (AFOs) and dynamic ankle foot orthoses (DAFOs) on gait in patients with spastic cerebral palsy (CP). Thirteen patients with dynamic equinus underwent motion analysis with electromyography. Both AFOs and DAFOs provided longer stride length, permitted pre-positioning for initial contact, and successfully controlled the excessive plantarflexion during the swing phase. Median frequency (MF) of EMG signal indicated that extremely high firing was found in the patient's lower limbs compared to controls that resulted in tiredness. The DAFOs allowed a significantly larger total ankle range of motion than the AFOs. However, AFOs significantly reduced the MF while DAFOs did not. The reduced MF seen when wearing AFOs suggested an improvement of walking endurance. The DAFO had the advantage of less restriction on ankle movement, which avoids muscular atrophy and improves orthotic compliance.

The psychometric properties of the Quantitative-Checklist for Autism in Toddlers (Q-CHAT) as a measure of autistic traits in a community sample of Singaporean infants and toddlers.

BACKGROUND: There is growing research evidence that subclinical autistic traits are elevated in relatives of individuals with autism spectrum disorder (ASD), continuously distributed in the general population and likely to share common etiology with ASD. A number of measures have been developed to assess autistic traits quantitatively in unselected samples. So far, the Quantitative-Checklist for Autism in Toddlers (Q-CHAT) is one of very few measures developed for use with toddlers as young as 18 months, but little is known about its measurement properties and factor structure. METHODS: The present study examined internal consistency, factor structure, test-retest stability, and convergent validity of the Q-CHAT in a sample of toddlers in Singapore whose caregivers completed the Q-CHAT at 18 (n = 368) and 24 months (n = 396). RESULTS: Three factors were derived accounting for 38.1 % of the variance: social/communication traits, non-social/behavioral traits, and a speech/language factor. Internal consistency was suboptimal for the total and speech/language scores, but acceptable for the social/communication and non-social/behavioral factor scores. Scores were generally stable between 18 and 24 months. Convergent validity was found with the Pervasive Developmental Disorders subscale of the Child Behavior Checklist (CBCL) completed by caregivers when their children were 24 months. Q-CHAT total scores in this sample were higher than those reported in other unselected samples from the UK. CONCLUSIONS: The Q-CHAT was found to have a three-factor structure, acceptable internal consistency for its two main factor scores (social/communication and non-social/behavioral), normally distributed scores in an unselected sample, and similar structure and measurement properties as those reported in other published studies. Findings are discussed in relation to existing literature and future directions for the validation of the Q-CHAT.

25-Hydroxycholecalciferol and insulin-like growth factor I are determinants of serum concentration of osteocalcin in elderly subjects with and without spinal fractures.

Osteocalcin is one of the major proteins in the osseous matrix. To evaluate the determinants and thus the physiological control of osteocalcin production in normal and osteoporotic subjects, the serum levels of osteocalcin, 25-hydroxycholecalciferol (25-OH-D3), and insulin-like growth factor I (IGFI) were measured by radioimmunoassay in 44 subjects over 60 years old. Circulating osteocalcin, 25-OH-D3, and IGFI were 0.28 +/- 0.10 nmol/L (1.65 +/- 0.96 ng/mL), 70.5 +/- 25.1 nmol/L (28.3 +/- 10.1 ng/mL), and 23.8 +/- 12.0 nmol/L in 27 healthy controls respectively, as compared with 0.09 +/- 0.09 nmol/L (0.52 +/- 0.52 ng/mL), 48.2 +/- 19.9 nmol/L (19.35 +/- 7.91 ng/mL), and 16.56 +/- 6.96 nmol/L in 17 patients with spinal fractures. Significant correlation was found between osteocalcin and 25-OH-D3 level, as well as between osteocalcin and IGFI. The results show that IGFI and 25-OH-D3 are important determinants for serum concentration of osteocalcin in elderly subjects with and without spinal fractures.

Vitamin D status among patients with fractured neck of femur in Hong Kong.

Vitamin D deficiency leads to secondary hyperparathyroidism initially and then to mild osteomalacia, both of which conditions may be aymptomatic and may predispose to bone fracture. To assess the importance of vitamin D deficiency in predisposing to fractured neck of femur, we studied the vitamin D status, dietary intake and socio-economic characteristics in 69 patients with fractured neck of femur (group A), 28 normal subjects with age above 60 (group B), and 101 normal volunteers (group C). Patients with fractured neck of femor had significantly lower levels of serum 25-hydroxy-cholecalciferol compared with subjects of groups B and C. There is no statistically significant difference in other biochemical parameters, including calcium, phosphate, and alkaline phosphatase. Patients with fractured neck of femur and with 25-hydroxycholecalciferol below 20 ng/mL were characterized by a home-bound and/or institutionalized life-style, smaller living place, and limited access to open space. To conclude, hypovitaminosis D is a common problem among elderly patients with fractured neck of femur in Hong Kong. The fact that such vitamin D deficiency is associated with muscle weakness may contribute to falls, and thus indirectly account for an increased rate of hip fractures over the normal control.

A linear accelerator couch attachment for extended SSD treatments.

An attachment to an existing linear accelerator couch is described that extends the range of SSD's at which patients can be treated. This couch attachment, which is mounted on castors for ease of mobility, is attached to and locks into the rail of the accelerator couch such that the axis of the new device is normal to the existing couch. All motions of this assembly are then under the control of the existing couch. The overall size of the couch attachment is 215 cm x 90 cm, sufficiently large to treat all clinically used fields.

Visualization of internal motion within a treatment portal during a radiation therapy treatment.

We offer a method for visualization of treatment portals continually during a radiation therapy treatment. Using digital image processing in conjunction with a fluorescent screen, portal images are obtained almost instantly. Images of quality that are clinically useful can be visualized every second. Digital enhancement further improves the quality of the images. The availability of instant portal images before each treatment can be most helpful in verifying that the patient is correctly and precisely set up prior to the onset of radiation. The ability to visualize treatment portals continuously during a treatment can help to ensure that radiation is restricted to the prescribed field and is not altered significantly by patient motion. In this report, we show and discuss a sequence of instant portal images obtained during a single treatment in which motion and absence of motion of internal anatomy is clearly demonstrated.

Three-dimensional volumetric assessment of response to treatment: stage I and II diffuse large cell lymphoma of the mediastinum.

From 1981 to 1986, 12 patients with Stage I and II diffuse large cell lymphoma of the mediastinum were treated with 4 or more cycles of multiagent chemotherapy and for nine patients this was followed by mediastinal irradiation. The response to treatment was assessed by three-dimensional volumetric analysis utilizing thoracic CT scans. The initial mean tumor volume of the five patients relapsing was 540 ml in contrast to an initial mean tumor volume of 360 ml for the seven patients remaining in remission. Of the eight patients in whom mediastinal lymphoma volumes could be assessed 1-2 months after chemotherapy prior to mediastinal irradiation, the three patients who have relapsed had volumes of 292, 92, and 50 ml (mean volume 145 ml) in contrast to five patients who have remained in remission with residual volume abnormalities of 4-87 ml (mean volume 32 ml). Four patients in prolonged remission with CT scans taken one year after treatment have been noted to have mediastinal tumor volumes of 0-28 ml with a mean value of 10 ml. This volumetric technique to assess the extent of mediastinal large cell lymphoma from thoracic CT scans appears to be a useful method to quantitate the amount of disease at presentation as well as objectively monitor response to treatment.

Strengthening mechanisms of bone bonding to crystalline hydroxyapatite in vivo.

The formation and strengthening mechanisms of bone bonding of crystalline hydroxyapatite (HA) has been investigated using high-resolution transmission electron microscope (HRTEM) and energy-dispersive X-ray (EDX) analysis. A series of results were obtained: (i) a layer of amorphous HA, which has almost the same chemistry as the implanted HA, was formed on the surface of crystalline HA particles prior to dissolution; (ii) at 3 months a bone-like tissue formed a bonding zone between mature bone and the HA implant, composed of nanocrystalline and amorphous apatite; and (iii) at 6 months, mature bone was in direct contact with HA particles, and collagen fibres were perpendicularly inserted into the surface layer of implanted HA crystals. Findings (i) and (ii) indicated the following dissolution-precipitation process. (i) The crystalline HA transforms into amorphous HA; (ii) the amorphous HA dissolves into the surrounding solution, resulting in over-saturation; and (iii) the nanocrystallites are precipitated from the over-saturated solution in the presence of collagen fibres. A preliminary analysis indicated several conclusions: (i) the transition from crystalline to amorphous HA might be the controlling step in the bone bonding of crystalline HA; (ii) biological interdigitation (or incorporation) of collagen fibres with HA and chemical bonding of a apatite layer were both necessary to strengthen and toughen a bone bond, not only for the bonding between bone and HA at 6 months, but also for the bonding zone at 3 months, which would otherwise be very fragile due to the inherited brittleness of polycrystalline ceramics; and (iii) perpendicular interdigitation is an effective way for collagen fibres to impart their unique combination of flexibility and strength to the interface which they are keying.

Loosening of sacral screw fixation under in vitro fatigue loading.

Sacral screw fixation is frequently used for fusion of the lower lumbar spine, but sacral screws appear to offer less secure fixation than lumbar pedicle screws, and failure due to loosening under fatigue loading is common. The aim of this study was to examine in vitro the stability of medial and lateral bicortical and unicortical sacral screw fixation under a physiologically relevant fatigue-loading pattern. Bone mineral density, screw insertion torque, and screw-fixation stiffness were measured prior to cyclic loading between 40 and 400 N compression at 2 Hz for 20,000 cycles. The screw-fixation stiffness was measured every 500 cycles, and the axial pullout strength of the screws was recorded following loading. All of the lateral insertions loosened under the applied loading, but some of the medial insertions remained stable. Medial insertions proved stiffer and stronger than lateral insertions, and bicortical fixations were stronger than unicortical fixations. Bone mineral density and insertion torque were correlated with screw stiffness and pullout strength, although better correlation was found for insertion torque than bone mineral density. Bone mineral density is a good preoperative indicator of sacral screw-fixation strength, and insertion torque is a good intraoperative indicator. An insertion torque greater than 1.5 Nm is suggested as an indicative value for a stable medial unicortical insertion, whereas an insertion torque greater than 2 Nm suggests a stable medial bicortical insertion. It appears that, apart from the choice of technique (screw orientation and depth), minimizing the load on the screws during the initial part of the fusion process is also critical to maintain stability of the fused section and to obtain a solid fusion mass.

Tissue phantom ratios for a Clinac 4/100.

Tissue phantom ratios (TPR), based on a normalization depth of 5 cm, have been measured in water for field sizes from 5 x 5 cm2 to approximately 40 x 40 cm2 and for depths from 1 to 40 cm for a Varian Clinac 4/100. These TPR's have been compared with those calculated from percent depth doses measured at the same time, and the two sets of data generally agree to better than 1%, with an average ratio of measured to calculated TPR of 0.999 +/- 0.013. Beam profiles have been measured for open and wedged fields, with particular concern for the often observed "horns," or the increase in dose at the corners of the field. The maximum dose at a depth of 1 cm, along the diagonal of the field for this machine, is approximately 5% higher than at the same depth on the central axis, whereas along the principal plane the maximum dose is only about 3% higher.

Ultrastructural study of mineralization of a strontium-containing hydroxyapatite (Sr-HA) cement in vivo.

The purpose of this study was to investigate the mineralization leading to osseointegration of strontium-containing hydroxyapatite (Sr-HA) bioactive bone cement injected into cancellous bone in vivo. Sr-HA cement was injected into the ilium of rabbits for 1, 3, and 6 months. The bone mineralization area was found to be largest at 3 months, then at 1 month, and smallest at 6 months (p < 0.01) measured with tetracycline labeling. Osseointegration of Sr-HA cement was achieved at 3 months as observed by scanning electron microscopy. A high calcium and phosphorus area was observed at the interface of bone-Sr-HA cement determined by energy-dispersive X-ray analysis. Transmission electron microscopy gave evidence of the mechanism of bone formation. Dissolution of Sr-HA into debris by the bone remodeling process was thought to increase the concentration of calcium and phosphorus at the interface of bone-Sr-HA cement and stimulate bone formation. Crystalline Sr-HA formed an amorphous layer and dissolved into the surrounding solution, then apatite crystallites were precipitated and formed new bone at 3 months. This young bone then becomes mature bone, which bonds tightly to the Sr-HA cement with collagen fibers inserted perpendicularly after 6 months.

In vivo cancellous bone remodeling on a strontium-containing hydroxyapatite (sr-HA) bioactive cement.

The purpose of this study was to investigate the in vivo bone response to the strontium-containing hydroxyapatite (Sr-HA) bioactive bone cement injected into the cancellous bone. Sr-HA cement was injected into the iliac crest of rabbits for 1, 3, and 6 months. Active bone formation and remodeling were observed after 1 month. Newly formed bone was observed to grow onto the bone cement after 3 months. Thick osteoid layer with osteoblasts formed along the bone and guided over the bone cement surface reflected the stimulating effect of Sr-HA. From scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) analysis, high calcium and phosphorus levels were detected at the interface with a thick layer of 70 microm in width, and fusion of Sr-HA with the bone was observed. Blood vessels were found developing in remodeling sites. The affinity of bone on Sr-HA cement was increased from 73.55 +/- 3.50% after 3 months up to 85.15 +/- 2.74% after 6 months (p < 0.01). In contrast to Sr-HA cement, poly(methyl methacrylate) (PMMA) bone cement was neither osteoconductive nor bioresorbable. Results show that the Sr-HA cement is biocompatible and osteoconductive, which is suitable for use in treating osteoporotic vertebral fractures.

The shoulder girdle in ankylosing spondylitis.

We report the clinical findings in the shoulders of fifty-two patients who had ankylosing spondylitis. The three major patterns that were found included limitation of scapulothoracic motion (forty-one patients), acute inflammatory arthropathy of the sternoclavicular or acromioclavicular joint (six patients), and severe restriction of glenohumeral motion (five patients). Although radiographic abnormalities were common, they correlated with the clinical findings in only minor respects.

Effusions in the knee in elderly patients who were operated on for fracture of the hip.

The incidence of an effusion in the knee in 155 consecutive elderly patients who had been operated on for a proximal femoral fracture was studied. The preoperative incidence had been 7.7 per cent in the ipsilateral knee and 1.3 per cent in the contralateral knee. Postoperatively, fifty patients (32.3 per cent) had an effusion on the ipsilateral side. In seven of them, the effusion had been present before the operation. All of the effusions subsided completely within three weeks after the operation. Results of the laboratory analysis of a specimen of the effused material from eight patients who were chosen at random showed non-inflammatory fluid. Probably the effusions were traumatic in origin, and it is likely that they were a response to stresses that had been incurred during the operation or at the time of fracture.

Changes in residual volume relative to vital capacity and total lung capacity after arthrodesis of the spine in patients who have adolescent idiopathic scoliosis.

We evaluated pulmonary functions before correction and again after a mean follow-up of three years in thirty-five patients who had adolescent idiopathic scoliosis. The mean age (and standard deviation) at the time of correction was 13.7 +/- 1.8 years, and at the time of follow-up it was 17.1 +/- 2.5 years. The findings in the patients were compared with those in matched normal control subjects. With the exception of forced vital capacity, all of the determinations of absolute pulmonary volume increased postoperatively, but the increases were not all proportional. When the preoperative and follow-up determinations were expressed as percentages of the predicted pulmonary volumes (on the basis of age) to eliminate any effects of the difference in age, there was no change in total lung capacity, but vital capacity and forced vital capacity were significantly reduced. In addition, there was a significant increase in residual volume. Of the mean increase in total lung capacity after correction of the scoliosis, 82 per cent was due to an increase in residual volume and 18 per cent, to an increase in vital capacity. However, in control subjects age-matched at the time of follow-up, the increase in vital capacity contributed 69 per cent of the mean increase in total lung capacity, a very marked difference from the findings in the patients who had scoliosis. In addition, two pulmonary-volume ratios--residual volume to vital capacity and residual volume to total lung capacity--increased in a highly significant fashion (Mann-Whitney test, p < 0.001) after arthrodesis of the spine.(ABSTRACT TRUNCATED AT 250 WORDS)

Surgical treatment of scoliosis following poliomyelitis. A review of one hundred and ten cases.

We reviewed the cases of 110 patients with paralytic scoliosis due to poliomyelitis who were operated on. In the lumbar region, anterior Dwyer instrumentation with posterior fusion gave excellent correction of scoliosis and pelvic obliquity. In the more rigid thoracic curves, combined anterior Dwyer instrumentation and posterior fusion gave better results than posterior fusion alone, but had more morbidity. Long c-shaped curves benefited more from a combined anterior Dwyer procedure at the apex of the curve and long posterior Harrington instrumentation. Traction was found to be of use only in rigid curves and in those larger than 80 degrees. With combined anterior and posterior fusion, there was a pseudarthrosis rate of 7 per cent in lumbar curves, none in thoracic curves, and 12.5 per cent in long c-shaped curves. The pseudarthrosis rate rose to more than 25 per cent in patients who had a posterior fusion alone.