Effects on the buoyant density of rabbit platelets of ADP and agents that increase the concentration of cyclic AMP

Rabbit platelets were aggregated by adenosine diphosphate (ADP), allowed to deaggregate and then separated into density subpopulations by centrifugation through discontinuous Stractan density gradients. Although ADP causes little or no release of the contents of the amine storage granules of rabbit platelets, ADP caused a decrease in platelet density as compared with control platelets subjected to the same procedures except for exposure to ADP. The density change persisted for at least four hours. The apparent size of platelets stimulated with ADP increased initially, but returned to control values during a one-hour period. A similar decrease in platelet density was observed with an albumin density gradient. Under conditions in which aggregation did not occur in response to ADP with ethylenediaminetetraacetic acid (EDTA) in the medium, little or no decrease in platelet density was observed. Agglutination with polylysine did not change platelet density. Thus, not only agents such as thrombin and plasmin that cause the release of the contents of the platelet granules decrease platelet density, but ADP also has this effect. Platelets would be exposed to all of these stimuli during thromboembolic processes, and their effect on platelets may account for the decrease in platelet density observed previously in experiments with rabbits with indwelling aortic catheters. Agents that increase the concentration of cyclic AMP (cAMP) in platelets (PGE1, adenosine, dibutyryl cAMP, forskolin, and papaverine) also decreased platelet density. This effect persisted when the platelets were washed and resuspended in fresh medium and was also demonstrable in plasma. Platelet size was gradually increased by prostaglandin E1 (PGE1) which maintains platelets in a disc shape and does not cause the release of granule contents, indicating that the decrease in platelet density caused by PGE1 may be attributable to platelet swelling.     

Autoantibodies inhibit interleukin-7-mediated proliferation and are associated with the age-dependent loss of pre-B cells in autoimmune New Zealand Black Mice

Surface IgM+B220+ B cell precursors can be categorized as either leukosialin (CD43/S7) negative (late stage pre-B cells) or positive (pro-B/early pre-B cells). In autoimmune New Zealand Black (NZB) mice, bone marrow small pre-B cells (IgM-CD43-B220+) and pro-B/early pre-B cells (IgM-CD43+B220+) declined significantly with age. In particular, subpopulations of pro-B/early pre-B cells expressing the heat stable antigen (HSA) were found in lower proportions with age. Significant decreases in interleukin-7 (IL-7) colony forming units (CFU) were also seen in NZB mice by 6 to 8 months of age and accompanied alterations in the numbers of pro-B and pre-B cells in bone marrow. Concomitant with reduced numbers of B lineage precursor cells and IL-7 CFU in vivo, NZB mice produced serum IgM antibodies that strongly inhibited IL-7 CFU responses in vitro. Two monoclonal IgM antibodies (5G9, 2F5) derived from LPS stimulated 10-month-old NZB splenocytes recognized pre-B cell surface antigens on both pre-B cell lines and on IL-7 stimulated bone marrow pro-B/pre-B cells. However, these monoclonal antibodies (MoAb) failed to significantly stain ex vivo bone marrow cells. The 5G9 and 2F5 MoAbs also partially inhibited IL-7 CFU in vitro. These results suggest that NZB bone marrow becomes increasingly deficient in B cell precursors and especially in IL-7 responsive pre-B cells with age. IgM serum antibodies and monoclonal IgM antibodies derived from older NZB mice inhibit pre-B cell growth to IL-7. The production of such autoantibodies may interfere with B cell development in aging NZB mice by preventing IL-7-mediated proliferation.     

Biological evaluation of intervertebral disc cells in different formulations of gellan gum‐based hydrogels

Gellan gum (GG)‐based hydrogels are advantageous in tissue engineering not only due to their ability to retain large quantities of water and provide a similar environment to that of natural extracellular matrix (ECM), but also because they can gelify in situ in seconds. Their mechanical properties can be fine‐tuned to mimic natural tissues such as the nucleus pulposus (NP). This study produced different formulations of GG hydrogels by mixing varying amounts of methacrylated (GG‐MA) and high‐acyl gellan gums (HA‐GG) for applications as acellular and cellular NP substitutes. The hydrogels were physicochemically characterized by dynamic mechanical analysis. Degradation and swelling abilities were assessed by soaking in a phosphate buffered saline solution for up to 170 h. Results showed that as HA‐GG content increased, the modulus of the hydrogels decreased. Moreover, increases in HA‐GG content induced greater weight loss in the GG‐MA/HA‐GG formulation compared to GG‐MA hydrogel. Potential cytotoxicity of the hydrogel was assessed by culturing rabbit NP cells up to 7 days. An MTS assay was performed by seeding rabbit NP cells onto the surface of 3D hydrogel disc formulations. Viability of rabbit NP cells encapsulated within the different hydrogel formulations was also evaluated by Calcein‐AM and ATP assays. Results showed that tunable GG‐MA/HA‐GG hydrogels were non‐cytotoxic and supported viability of rabbit NP cells. Copyright © 2012 John Wiley & Sons, Ltd.     

Rectal biopsy in patients presenting to an infectious disease unit with diarrhoeal disease.

The role of sigmoidoscopy and rectal biopsy was investigated in patients referred to an infectious diseases unit with diarrhoea. Seventy-four patients were studied. Nine patients (12%) had inflammatory bowel disease, either ulcerative colitis or Crohn's disease. Thirty-six patients (48%) had infective diarrhoea. A wide variety of conditions accounted for the diarrhoea in the remaining patients. Sigmoidoscopy was abnormal in 25 patients and rectal biopsy in 56. The abnormalities in rectal mucosal histology were classified into six grades. Some patients with infective diarrhoea showed rather characteristic histological changes which may be of diagnostic value. Eight showed features which suggested a diagnosis of inflammatory bowel disease. However, repeat rectal biopsy in the convalescent period showed a striking improvement in the patients with infective diarrhoea. In contrast, the histological changes persisted in the patients with inflammatory bowel disease. Repeat rectal biopsy may be essential before making a firm diagnosis of inflammatory bowel disease in some patients who present with diarrhoea and apparently typical histological changes.     

Circadian variation in ventricular electrical instability associated with coronary artery disease

Although sudden cardiac deaths and ischemic cardiac events clearly occur in a circadian pattern, such a pattern has not been shown for primary arrhythmic events. Because primary arrhythmic events are thought to play an important role in sudden cardiac death, a large series of ventricular stimulation studies was analyzed to determine whether circadian variation in ventricular electrical instability exists. If such a circadian variation could be shown, it could have implications for the conduct and interpretation of electrophysiologic testing and the etiology of circadian variation in sudden cardiac death. Results of 2 drug-free ventricular stimulation studies performed 4 to 28 hours apart in each of 162 patients with coronary artery disease were analyzed. Rate and duration of induced arrhythmia, number of extrastimuli required to induce arrhythmia and changes in these factors between the 2 tests in each patient were analyzed. Comparisons were made by half-day, by hour and in a temporally continuous manner to eliminate errors associated with any single method. No significant circadian variation was found in any electrophysiologic measure of ventricular electrical instability despite adequate statistical power. These findings show that the time of day during which ventricular stimulation tests are performed does not affect test results, and therefore does not need to be controlled during electrophysiologic studies. If these findings are parallel to those in ambulatory patients with coronary artery disease, then circadian changes in ventricular electrical instability may not play as important a role in the circadian pattern of sudden cardiac death as had been previously thought.     

Reliability of measuring hip abductor strength following total knee arthroplasty using a hand-held dynamometer

Purpose: To investigate the test–retest reliability of measuring hip abductor strength in patients with total knee arthroplasty (TKA) using a hand-held dynamometer (HHD) with two different types of resistance: belt and manual resistance. Method: Test–retest reliability of 30 subjects (17 female, 13 male, 71.9?±?7.4 years old), 9.2?±?2.7 days post TKA was measured using belt and therapist resistance. Retest reliability was calculated with intra-class coefficients (ICC_3,1) and 95% confidence intervals (CI) for both the group average and the individual scores. A paired t-test assessed whether a difference existed between the belt and therapist methods of resistance. Results: ICCs were 0.82 and 0.80 for the belt and therapist resisted methods, respectively. Hip abductor strength increases of 8?N (14%) for belt resisted and 14?N (17%) for therapist resisted measurements of the group average exceeded the 95% CI and may represent real change. For individuals, hip abductor strength increases of 33?N (72%) (belt resisted) and 57?N (79%) (therapist resisted) could be interpreted as real change. Conclusions: Hip abductor strength can be reliably measured using HHD in the clinical setting with the described protocol. Belt resistance demonstrated slightly higher test–retest reliability.

• Implications for Rehabilitation

• Reliable measurement of hip abductor muscle strength in patients with TKA is important to ensure deficiencies are addressed in rehabilitation programs and function is maximized.

• Hip abductor strength can be reliably measured with a hand-held dynamometer in the clinical setting using manual or belt resistance.

     

The Montreal Neurological Institute: training of the first African-American neurosurgeons

BACKGROUND: Since its inception in 1934 by the legendary Dr. Wilder Penfield, the Montreal Neurological Institute (MNI) has provided world-renowned instruction in neurosurgery and related neurosciences, training many of the most prominent figures in the history of neurosurgery. Less well known is the role of the MNI in training the first African-American board-certified neurosurgeons. METHODS: A comprehensive review of pertinent modern and historical records spanning the past century was performed. RESULTS: From 1947-1965, the MNI trained the first African-American board-certified neurosurgeon, and three of the first four. The first, Dr. Clarence Greene, Sr., trained at MNI from 1947-1949. The next, Dr. Jesse Barber, Jr., trained at MNI from 1958-1961. Like Greene, Barber received his MD from the Howard University College of Medicine, was on the general surgery faculty at Howard before training at MNI under Penfield and returned to Howard following his training. The third, Dr. Lloyd Dayes, matriculated at MNI in 1960 after receiving his MD from the Loma Linda University School of Medicine and trained from 1961-1965 under Dr. Theodore Rasmussen, after which he returned to Loma Linda. Greene, Barber and Dayes were certified by the American Board of Neurological Surgery in 1953, 1963 and 1967, respectively, as the first, third and fourth African-American neurosurgeons. CONCLUSION: The willingness of the world-renowned MNI to train the first African-American neurosurgeons during a time of intense racial segregation in the United States played a major role in enabling subsequent African Americans to enter and enhance the field of neurosurgery.