The Reaction of three Purified Lectins with Guinea Pig Lymphocytes

Three purified phytohemagglutinins, LcH-A, LcH-B, and AbH bind to guinea pig lymphocytes, the first two stimulating lymphocytes in culture and the last causing no stimulaton. By fluorescence microscopy essentially every examined lymphocyte isolated from various lymphoid organs bound both LcH-A and AbH. From the fact that both the stimulant (LcH-A) and the nonstimulant (AbH) showed similar microscopic staining patterns, it is suggested that the type of staining pattern seen is not directly related to the stimulation process. Although LcH-A and LcH-B have very similar specificities, they have subtly different lymphocyte stimulating properties, the former being a better stimulant for blood and lymph node cells and the latter a better stimulant of thymus cells. Despite the fact that AbH is not a stimulator of guinea pig lymphocytes, it could, under certain conditions, significantly affect the stimulation of guinea pig lymphocytes by LcH-A.     

Effect of growth factors on cell proliferation, matrix deposition, and morphology of human nasal septal chondrocytes cultured in monolayer

OBJECTIVES: Tissue engineering of septal cartilage provides ex vivo growth of cartilage from a patient's own septal chondrocytes for use in craniofacial reconstruction. To become clinically applicable, it is necessary to rapidly expand a limited population of donor chondrocytes and then stimulate the production of extracellular matrix on a biocompatible scaffold. The objective of this study was to determine favorable serum-free culture conditions for proliferation of human septal chondrocytes using various concentrations and combinations of four growth factors. STUDY DESIGN: Prospective, randomized, controlled study. METHODS: Nasal septal chondrocytes from six patient donors were isolated by enzymatic digestion and expanded in monolayer culture in both serum-free media (SFM) and 2% fetal bovine serum (FBS). Both of these groups were exposed to varying concentrations and combinations of transforming growth factor (TGF)-beta1, basic fibroblast growth factor (FGF)-2 both at 1, 5, and 25 ng/mL, and bone morphogenetic protein (BMP)-2 and insulin-like growth factor (IGF)-1, both at 5, 25, and 125 ng/mL in the medium during the expansion phase. Cell morphology was assessed throughout the culture duration. After 7 days of monolayer growth, cultures were assessed for cellularity and glycosaminoglycan (GAG) content. RESULTS: The addition of low-dose FBS in culture media consistently led to significantly greater cell proliferation and matrix deposition than the SFM cell cultures. FGF-2 and TGF-beta1 both alone and in combination led to the greatest proliferative effect compared with the other growth factors. In contrast, BMP-2 and IGF-1 led to the least cell proliferation although was most effective in retaining chondrocyte cell morphology. CONCLUSIONS: With the addition of TGF-beta1 and FGF-2 to culture media, the concentration of serum can be greatly decreased and possibly eliminated altogether without jeopardizing cell proliferation.     

Development of UV-induced squamous cell carcinomas is suppressed in the absence of SPARC

SPARC (Secreted Protein Acidic and Rich in Cysteine) is a multifunctional glycoprotein belonging to a group of matrix-associated factors that mediate cell-extracellular matrix interactions but have no structural roles. In the present study we investigated the contribution of SPARC to factors that influence the development of skin tumors in response to UV irradiation. A hairless SPARC-null mouse was developed and compared to control SKH1 hairless mice in terms of skin tumor induction and extracellular matrix changes occurring in response to UV-irradiation. Following 23 weeks of exposure to UVB totaling 14.5 J per cm(2), tumor development in the wild-type mice was severe, with an average of over 20 tumors per mouse, many of which were squamous cell carcinomas. Conversely, the SPARC-null mice were strikingly tumor-resistant, developing no squamous cell carcinomas and averaging less than one small papilloma per mouse. SPARC was undetectable immunohistochemically in skin from the non-irradiated control group yet was present in relatively high quantities in the basal and superficial areas of the tumor mass. The SPARC-null mice also exhibited a limited contact hypersensitivity response and were refractory to UV induced immune suppression. In conclusion, SPARC appears to have a crucial role in mediating tumor formation in response to UV irradiation.     

Deficits in the evolution of hand preshaping in Parkinson's disease

Parkinson's disease (PD) results in various types of motor impairments including bradykinesia, tremor and rigidity. Recent research has implicated more fundamental processes at the source of the observed motor deficits. Among these, problems in the sequencing and/or timing of complex movements and in the execution of internally-guided tasks. Furthermore, PD patients exhibit procedural learning deficits which may complicate the interpretation of experimental results of studies involving novel sensorimotor tasks. The reach-to-grasp movement is a complex, overlearned sensorimotor task consisting of two semi-independent components, a relatively simple reach or transport phase and a more complex manipulation or prehension phase. In the present study, we used a novel technique in order to study the evolution of hand preshaping during the reach-to-grasp movement of PD patients and age-matched controls to objects of different shapes in three different spatial locations. Our results indicate that while PD patients are able to specify movement direction as well as controls, their hand preshaping exhibits substantial impairments. Other prehension measures, such as the time to peak aperture (TPA), indicate that PD patients delayed execution of the grasp until visual feedback of their hand was available. Overall, our results suggest that PD patients' internal guidance processes are severely disrupted, having to rely on visual feedback in order to modulate their hand shape to fit the contours of the target objects during a reach-to-grasp movement.     

Amelioration of diabetic nephropathy in SPARC-null mice

SPARC (Secreted Protein, Acidic and Rich in Cysteine) is a matricellular protein that inhibits mesangial cell proliferation and also affects production of extracellular matrix (ECM) by regulating transforming growth factor-beta1 (TGF-beta1) and type I collagen in mesangial cells. This study is an investigation of the role of SPARC in streptozotocin (STZ)-induced diabetic nephropathy (DN) of 6-mo duration in wild type (WT) and SPARC-null mice. SPARC expression was evaluated by immunohistochemistry (IHC) and by in situ hybridization (ISH). Deposition of type I and IV collagen and laminin was evaluated by IHC, and TGF-beta 1 mRNA was assessed by ISH. Renal function studies revealed no significant difference in BUN between diabetic SPARC-null mice and diabetic WT mice, whereas a significant increase in albumin excretion was detected in diabetic WT relative to diabetic SPARC-null mice. Diabetic WT animals exhibited increased levels of SPARC mRNA and protein in glomerular epithelial cells and in interstitial cells, in comparison with nondiabetic WT mice. Neither SPARC mRNA nor protein was detected in SPARC-null mice. Morphometry revealed a significant increase in the percentage of the glomerular tufts occupied by ECM in diabetic WT compared with nondiabetic WT mice, although there was no difference in the mean glomerular tuft area among groups. In contrast, diabetic SPARC-null mice did not show a significant difference in the percentage of the glomerular tufts occupied by ECM relative to nondiabetic null mice. Tubulointerstitial fibrosis was ameliorated in diabetic SPARC-null mice compared with diabetic WT animals. Further characterization of diabetic SPARC-null mice revealed diminished glomerular deposition of type IV collagen and laminin, and diminished interstitial deposition of type I and type IV collagen correlated with decreases in TGF-beta 1 mRNA compared with WT diabetic mice. These observations suggest that SPARC contributes to glomerulosclerosis and tubulointerstitial damage in response to hyperglycemia through increasing TGF-beta 1 expression in this model of chronic DN.     

Immunolocalization of ClC-K chloride channel in strial marginal cells and vestibular dark cells

Secretion of K⁺ into endolymph depends on a particular constellation of ion transport proteins in the apical and basolateral membranes of strial marginal cells and vestibular dark cells. One fundamental component is the large chloride conductance of the basolateral membrane, which recycles chloride taken up by the Na⁺-K⁺-Cl⁻ cotransporter in the same membrane. Evidence has been reported recently that supports ClC-K, a channel subunit previously thought to be specific to the kidney, as being the molecular entity underlying this conductance. We have isolated protein from the gerbil kidney, stria vascularis and vestibular labyrinth and found by Western blot analysis a 60 kDa band, a 48 kDa band and 54 and 70 kDa bands, respectively, specifically labeled by ClC-K antibody. Subsequent immunohistochemical observations of the inner ear tissues with a confocal microscope on fluorescently labeled tissue sections showed the staining to be restricted to the basolateral region of strial marginal cells and vestibular dark cells. The cochlear staining was distinct from the distribution of the Kir4.1 (KCNJ10) K⁺ channel, known to be present only in strial intermediate cells. These findings support the contention that ClC-K is an important component of the basolateral Cl⁻ conductance that participates in K⁺ secretion by these epithelia.     

Cavernous haemangiomas (angiomas) of the brain

This paper highlights the importance of cavernous haemangiomas as clinically significant lesions and the role of imaging, particularly MRI, in suggesting the diagnosis. An understanding of the pathology of these lesions helps to explain the features demonstrated by imaging techniques.