General care in the management of severe traumatic brain injury: Latin American consensus. / Cuidados generales en el manejo del traumatismo craneoencefálico grave: consenso latinoamericano.

Severe traumatic brain injury (sTBI) remains prevalent in the young adult population. Indeed, far from descending, the incidence of sTBI remains high. One of the key bases of treatment is to avoid, detect and correct secondary injuries of systemic origin, which aggravate the primary lesion. Much of this can be achieved by maintaining an adequate physiological microenvironment allowing recovery of the damaged brain tissue. General care measures are nonspecific actions designed to meet that objective. The available guidelines on the management of sTBI have not included the topics contemplated in this consensus. In this regard, a group of members of the Latin American Brain Injury Consortium (LABIC), involved in the different aspects of the acute management of sTBI (neurosurgeons, intensivists, anesthesiologists, neurologists, nurses and physiotherapists) were gathered. An exhaustive literature search was made of selected topics in the LILACS, PubMed, Embase, Scopus, Cochrane Controlled Register of Trials and Web of Science databases. To establish recommendations or suggestions with their respective strength or weakness, the GRADE methodology (Grading of Recommendations, Assessment, Development and Evaluation) was applied. Additionally, certain recommendations (included in complementary material) were not assessed by GRADE, because they constitute a set of therapeutic actions of effective compliance, in which it was not possible to apply the said methodology. Thirty-two recommendations were established, 16 strong and 16 weak, with their respective levels of evidence. This consensus attempts to standardize and establish basic general care measures in this particular patient population.

Malignant hemispheric infarction of the middle cerebral artery. Diagnostic considerations and treatment options. / Infarto hemisférico maligno de la arteria cerebral media. Consideraciones diagnósticas y opciones terapéuticas.

INTRODUCTION: Malignant hemispheric infarction (MHI) is a specific and devastating type of ischemic stroke. It usually affects all or part of the territory of the middle cerebral artery although its effects may extend to other territories as well. Its clinical outcome is frequently catastrophic when only conventional medical treatment is applied. OBJECTIVE: The purpose of this review is to analyse the available scientific evidence on the treatment of this entity. DEVELOPMENT: MHI is associated with high morbidity and mortality. Its clinical characteristics are early neurological deterioration and severe hemispheric syndrome. Its hallmark is the development of space-occupying cerebral oedema between day 1 and day 3 after symptom onset. The mass effect causes displacement, distortion, and herniation of brain structures even when intracranial hypertension is initially absent. Until recently, MHI was thought to be fatal and untreatable because mortality rates with conventional medical treatment could exceed 80%. In this unfavourable context, decompressive hemicraniectomy has re-emerged as a therapeutic alternative for selected cases, with reported decreases in mortality ranging between 15% and 40%. CONCLUSIONS: In recent years, several randomised clinical trials have demonstrated the benefit of decompressive hemicraniectomy in patients with MHI. This treatment reduces mortality in addition to improving functional outcomes.

Aislamiento y rendimiento del GMP mediante precipitación de lactosuero con ácido tricloroacético / GMP isolation and yield by whey precipitation with trichloroacetic acid

Durante la elaboración del queso, la k-caseína es hidrolizada por la renina (Quimosina E.C.3.4.23.4) en el enlace peptídico Fen105-Met106 generando dos fracciones: la para-k-caseína y el glicomacropéptido (GMP) que se libera al lactosuero. El GMP presenta una estructura química particular donde predominan los aminoácidos con cadena lateral ramificada, no presenta aminoácidos aromáticos y contiene carbohidratos unidos a residuos de treonina; por esta razón se le ha atribuido una variedad de actividades biológicas. Se ha estimado que en Venezuela se generan alrededor de 713 toneladas de lactosuero anualmente. Un volumen considerable de este subproducto se produce en el estado Zulia, constituyéndose esto en una fuente de péptidos y proteínas de alta calidad nutricional que está siendo subutilizada. Con el propósito de evaluar el aislamiento y rendimiento del GMP a partir de la precipitación de lactosuero de ricotta con ácido tricloroacético 50 por ciento, se realizaron 6 extracciones con este ácido a 50 mL de cada tipo de suero analizado: suero ricotta, suero comercial resuspendido y suero ácido (control negativo). Se verificó mediante pruebas químicas y PAGE-SDS 15 por ciento de manera indirecta, la presencia de GMP en las preparaciones obtenidas. Se observaron bandas de 6,5; 18,3 y 19,0 kDa en suero ricotta y suero comercial resuspendido. Las bandas de 18,3 y 19,0 posiblemente correspondan a la forma trimérica del péptido. El rendimiento del GMP en términos de proteínas fue en promedio 1,17 mg/50mL (1,17 por ciento) y 4,51 mg/50mL (0,81 por ciento), para suero ricotta y suero comercial, respectivamente. Los resultados indican que es factible obtener preparaciones del GMP, sin embargo, para plantear la producción a escala industrial de este péptido para su aprovechamiento, se requiere evaluar otros procedimientos donde se obtenga a bajo costo una preparación purificada del GMP.

During cheese manufacturing k-casein is hydrolyzed by rennin (Quimosine E.C.3.4.23.4) on peptidic bond Fen105- Met106 releasing two fractions: para-k-casein and glycomacropeptide (GMP). GMP shows a particular chemical structure in which ramified lateral chain aminoacids prevail, without aromatics aminoacids, but with carbohydrates short chains linked to some threonine residues; because of this, a variety of biological activities have been attributed to molecule. It has been considered that in Venezuela, 713 tons of whey are generated annually. An important volume of this byproduct is produced in the Zulia State, becoming itself a source of peptides and proteins of high nutritional quality that has been subused. With the purpose of evaluating GMP isolation and yield from ricotta whey precipitation with 50 percent trichloroacetic acid treatment, 6 extractions where performed with this acid to 50 mL of each analyzed whey: ricotta whey, resuspended commercial whey and acid whey (negative control). By means of chemical tests and PAGE-SDS 15 percent, indirect presence of GMP was verified in all preparations. Bands of 6.5, 18.3 and 19.0 kDa were observed in ricotta and commercial whey. Bands of 18.3 and 19.0 possibly correspond to the peptide trimeric structure. GMP yield in terms of protein content was 1.17 mg/50mL (1.17 percent) and 4.51 mg/50mL (0.81 percent), for ricotta and commercial whey, respectively. Results show that it feasible to obtain preparations of GMP, however, in order to produce this peptide industrially for its use, evaluation of low cost procedures for GMP purification is required.

Creatinine clearance and adverse hospital outcomes in patients with acute coronary syndromes: findings from the global registry of acute coronary events (GRACE).

OBJECTIVE: To determine whether creatinine clearance at the time of hospital admission is an independent predictor of hospital mortality and adverse outcomes in patients with acute coronary syndromes (ACS). DESIGN: A prospective multicentre observational study, GRACE (global registry of acute coronary events), of patients with the full spectrum of ACS. SETTING: Ninety four hospitals of varying size and capability in 14 countries across four continents. PATIENTS: 11 774 patients hospitalised with ACS, including ST and non-ST segment elevation acute myocardial infarction and unstable angina. MAIN OUTCOME MEASURES: Demographic and clinical characteristics, medication use, and in-hospital outcomes were compared for patients with creatinine clearance rates of > 60 ml/min (normal and minimally impaired renal function), 30-60 ml/min (moderate renal dysfunction), and < 30 ml/min (severe renal dysfunction). RESULTS: Patients with moderate or severe renal dysfunction were older, were more likely to be women, and presented to participating hospitals with more comorbidities than those with normal or minimally impaired renal function. In comparison with patients with normal or minimally impaired renal function, patients with moderate renal dysfunction were twice as likely to die (odds ratio 2.09, 95% confidence interval 1.55 to 2.81) and those with severe renal dysfunction almost four times more likely to die (odds ratio 3.71, 95% confidence interval 2.57 to 5.37) after adjustment for other potentially confounding variables. The risk of major bleeding episodes increased as renal function worsened. CONCLUSION: In patients with ACS, creatinine clearance is an important independent predictor of hospital death and major bleeding. These data reinforce the importance of increased surveillance efforts and use of targeted intervention strategies in patients with acute coronary disease complicated by renal dysfunction.

A Cbfa1-dependent genetic pathway controls bone formation beyond embryonic development.

The molecular mechanisms controlling bone extracellular matrix (ECM) deposition by differentiated osteoblasts in postnatal life, called hereafter bone formation, are unknown. This contrasts with the growing knowledge about the genetic control of osteoblast differentiation during embryonic development. Cbfa1, a transcriptional activator of osteoblast differentiation during embryonic development, is also expressed in differentiated osteoblasts postnatally. The perinatal lethality occurring in Cbfa1-deficient mice has prevented so far the study of its function after birth. To determine if Cbfa1 plays a role during bone formation we generated transgenic mice overexpressing Cbfa1 DNA-binding domain (DeltaCbfa1) in differentiated osteoblasts only postnatally. DeltaCbfa1 has a higher affinity for DNA than Cbfa1 itself, has no transcriptional activity on its own, and can act in a dominant-negative manner in DNA cotransfection assays. DeltaCbfa1-expressing mice have a normal skeleton at birth but develop an osteopenic phenotype thereafter. Dynamic histomorphometric studies show that this phenotype is caused by a major decrease in the bone formation rate in the face of a normal number of osteoblasts thus indicating that once osteoblasts are differentiated Cbfa1 regulates their function. Molecular analyses reveal that the expression of the genes expressed in osteoblasts and encoding bone ECM proteins is nearly abolished in transgenic mice, and ex vivo assays demonstrated that DeltaCbfa1-expressing osteoblasts were less active than wild-type osteoblasts. We also show that Cbfa1 regulates positively the activity of its own promoter, which has the highest affinity Cbfa1-binding sites characterized. This study demonstrates that beyond its differentiation function Cbfa1 is the first transcriptional activator of bone formation identified to date and illustrates that developmentally important genes control physiological processes postnatally.

Osteopontin (OPN) distribution in premalignant and malignant lesions of oral epithelium and expression in cell lines derived from squamous cell carcinoma of the oral cavity.

The objectives of this study were to assess the immunolocalization of human osteopontin (OPN) in oral lesions and to identify human cell lines of oral squamous cell carcinoma (OSCC) origin that express OPN mRNA. OPN was localized using immunohistochemistry in the following oral specimens: normal epithelium (n=6), epithelial hyperplasia (n=4), epithelial dysplasia (n=28), carcinoma in situ (n=11) and squamous cell carcinoma (n=43). Cell lines UMSCC-1, MDA TU 138, MDA 686LN, SCC4, SCC9, SCC25, CAL 27 and MDA 1483 were characterized for OPN mRNA expression using Northern blotting. OPN was not detected in normal oral epithelium. Intracellular and intercellular immunoreactivity was seen in 75% of hyperplasias, 57% of dysplasias, 54% of carcinoma in situ and 67% of squamous cell carcinomas. UMSCC-1 expressed high levels of OPN mRNA. We conclude that OPN protein is detectable in premalignant and malignant lesions arising from oral epithelium. UMSCC-1 may be a useful cell line in which to conduct in vitro studies designed to clarify the role of OPN in OSCC.

Expression and localization of PG-Lb/epiphycan during mouse development.

We have examined the expression pattern of the PG-Lb/epiphycan gene that encodes a small leucine-rich repeat proteoglycan during mouse embryonic development. PG-Lb/epiphycan mRNA transcripts were first detected at E12.5 days postcoitus (dpc) at high levels in structures that were developing cartilage elements. The gene is expressed in a very specific temporal and spatial fashion in cartilaginous structures. To examine PG-Lb/epiphycan gene expression during cartilage development in more detail, we performed in situ hybridization on hindlimb sections at specific stages of mouse embryonic development. The expression of PG-Lb/epiphycan was compared to that of collagen type II and collagen type X, which are early and late markers for cartilage development, respectively. The expression of PG-Lb/epiphycan occurs later than collagen type II in cartilage development, but its expression appears in the growth plate before and is excluded from the zone of hypertrophic chondrocytic cells expressing collagen type X. An antibody against PG-Lb/epiphycan localized the protein within the entire growth plate of the E17.5 dpc embryonic hindlimb cartilage including the hypertrophic zone where PG-Lb/epiphycan gene expression is turned off. Our results show that PG-Lb/epiphycan gene expression is an intermediate marker for chondrogenesis, and that the protein can be localized to the extracellular matrix surrounding resting, proliferating, and hypertrophic chondrocytes by immunofluorescence histochemistry.

Improved immunohistochemical staining of osteopontin (OPN) in paraffin-embedded archival bone specimens following antigen retrieval: anti-human OPN antibody recognizes multiple molecular forms.

Studies to assess osteopontin (OPN) localization in adult human bone using immunochemical techniques produce conflicting results due to variations in tissue processing or antibody immunoreactivity. The present study was designed to resolve these discrepancies using well-characterized antibodies and improved antigen detection. An anti-osteopontin (alpha-OPN) antiserum was developed that recognizes various soluble molecular weight forms of human OPN, including monomeric, cleaved, and dimerized products. An affinity column of full length recombinant human OPN (rOPN) coupled to support was used to purify alpha-OPN antibodies. Western analysis showed that the affinity-purified antibodies recognized numerous molecular weight forms of OPN. These antibodies were used to study the distribution of OPN in adult human bone using immunohistochemical techniques combined with an antigen retrieval protocol utilizing a newly developed antigen retrieval solution, Retriev-Alltrade mark (Bronco Technologies Inc, Pasadena, TX). Immunolocalization of OPN in archival bone specimens prior to antigen retrieval produced no demonstrable immunostaining even at high concentrations of alpha-OPN. Use of the antigen retrieval protocol restored OPN immunoreactivity, with strong staining apparent in cement lines, osteoblasts, osteocytes, canaliculi, osteoid, and bone matrix. We conclude that antigen retrieval restores immunochemical recognition of OPN in archival specimens containing bone without increasing nonspecific binding.

Spontaneous calcification of arteries and cartilage in mice lacking matrix GLA protein.

Calcification of the extracellular matrix (ECM) can be physiological or pathological. Physiological calcification occurs in bone when the soft ECM is converted into a rigid material capable of sustaining mechanical force; pathological calcification can occur in arteries and cartilage and other soft tissues. No molecular determinant regulating ECM calcification has yet been identified. A candidate molecule is matrix GLA protein (Mgp), a mineral-binding ECM protein synthesized by vascular smooth-muscle cells and chondrocytes, two cell types that produce an uncalcified ECM. Mice that lack Mgp develop to term but die within two months as a result of arterial calcification which leads to blood-vessel rupture. Chondrocytes that elaborate a typical cartilage matrix can be seen in the affected arteries. Mgp-deficient mice additionally exhibit inappropriate calcification of various cartilages, including the growth plate, which eventually leads to short stature, osteopenia and fractures. These results indicate that ECM calcification must be actively inhibited in soft tissues. To our knowledge, Mgp is the first inhibitor of calcification of arteries and cartilage to be characterized in vivo.

Integrated processes responsible for soft tissue healing.

Wounded soft tissues undergo repair through a complex series of interrelated events that involve both physical and chemical activities. These processes are currently undergoing extensive investigation as efforts are directed toward achieving augmented and accelerated healing. Early wound-healing research focused on expanding traditional histologic descriptions of tissue healing by attempting to characterize the environment and biologic mediators responsible for healing. These initial studies successfully identified a number of agents and physiochemical factors present in healing wounds, but their precise roles and importance remain largely unknown. This review article summarizes the current literature on soft tissue healing. An effort has been made to correlate the activities of the major growth factors and cytokines with the individual reparative processes including the inflammatory response, hemostasis, fibroplasia, angiogenesis, and remodeling. Explanations and characteristics of growth factor function as well as brief descriptions of several major factors and their spectrum of activity are also provided.

Increased bone formation in osteocalcin-deficient mice.

Vertebrates constantly remodel bone. The resorption of preexisting bone by osteoclasts and the formation of new bone by osteoblasts is strictly coordinated to maintain bone mass within defined limits. A few molecular determinants of bone remodelling that affect osteoclast activity have been characterized, but the molecular determinants of osteoblast activity are unknown. To investigate the role of osteocalcin, the most abundant osteoblast-specific non-collagenous protein, we have generated osteocalcin-deficient mice. These mice develop a phenotype marked by higher bone mass and bones of improved functional quality. Histomorphometric studies done before and after ovariectomy showed that the absence of osteocalcin leads to an increase in bone formation without impairing bone resorption. To our knowledge, this study provides the first evidence that osteocalcin is a determinant of bone formation.

Treatment of a patient with hereditary gingival fibromatosis: a case report.

A case of hereditary gingival fibromatosis is presented. Treatment consisted of apically positioned flap surgery and CO2 laser evaporation. Diagnostic and treatment issues are discussed.

Bone matrix proteins in osteogenesis and remodelling in the neonatal rat mandible as studied by immunolocalization of osteopontin, bone sialoprotein, alpha 2HS-glycoprotein and alkaline phosphatase.

The neonatal rat mandible was used as a model to study bone formation, mineralization, quiescence, and resorption, using immunolocalization and a variety of tissue-processing techniques. Monospecific antibodies for osteopontin (OPN), bone sialoprotein (BSP), alkaline phosphatase (AP) and alpha 2HS-glycoprotein (alpha 2HS-GP) were used on fixed paraffin-embedded tissue, fixed frozen tissue and unfixed frozen tissue. Immunostaining was correlated with mineral content by two procedures, the von Kossa and the morin techniques. Morin fluorescence was used with secondary immunostaining to provide a way of closely correlating bone matrix proteins and matrix mineralization. Co-immunolocalization procedures were used to compare the sites of bone proteins in the matrix. AP was found earliest during osteogenic cell differentiation, appearing in the preosteoblasts, followed by OPN and BSP, which first appeared in osteoblasts. alpha 2HS-GP expression was not observed in cells. The results provide clear evidence for the presence of OPN in osteoid, while BSP and alpha 2HS-GP were confined to the mineralized matrix. Immunostaining of bone proteins is highly technique-dependent: immunolocalization investigations required several methods of approach to ensure adequate demonstration of these proteins in cells and matrix. The results support the contention that osteopontin is multifunctional in bone metabolism, and that alpha 2HS-GP, though produced in the liver, is abundant in bone matrix and may also have a function in bone metabolism.

Molecular analysis of rat dentin sialoprotein.

Dentin sialoprotein (DSP) is a noncollagenous protein originally isolated from rat dentin. Because it is made by odontoblasts that are actively synthesizing dentin. DSP may play an important role in dentinogenesis. We have isolated a full length DSP cDNA from a rat odontoblast/dental pulp cDNA library (Ritchie et al. [1994] J. Biol. Chem. 269:3698-3702) which codes for a 17 residue signal peptide and a 366 residue, 53 kDa mature protein. In situ hybridization revealed DSP mRNA expression by odontoblasts, but no other cells, in jaws from newborn rat. Northern analysis of various rat tissues demonstrated the presence of DSP transcripts in newborn tooth germs and 21 day old rat incisors. Moreover, multiple transcripts of 4.6 kb and 1.5 kb were found in these two tissues. To better understand the origin of these DSP mRNA multiple transcripts, we have isolated two rat genomic clones. Digestion of each clone with EcoRI followed by Southern analysis revealed that DSP cDNA hybridized to a 4 kb fragment in a lambda dash clone and to a 6 kb fragment in a cosmid clone. Since DSP cDNA hybridized to a 6 kb EcoRI fragment and a 4 kb EcoRI fragment obtained from a rat liver genomic cDNA digested with EcoRI, the multiple DSP mRNA transcripts are most likely derived from two related DSP genes which coexist in the rat genome.

Normal long bone growth and development in type X collagen-null mice.

To investigate the role of type X collagen in skeletal development, we have generated type X collagen-null mice. Surprisingly, mice without type X collagen were viable and fertile and had no gross abnormalities in long bone growth or development. No differences were detected between the type X collagen-null mice and controls when growth plates of both newborn and 3-week old mice were examined by histology and by immunostaining for extracellular matrix components of bone including osteopontin, osteocalcin and type II collagen. Our results suggest that type X collagen is not required for long bone development. However, mice and humans with dominant acting type X collagen mutations have bone abnormalities, suggesting that only the presence of abnormal type X collagen can modify bone growth and development.

Immunolocalization of connexin 43 in the tooth germ of the neonatal rat.

Rabbit polyclonal antibodies to amino acids 346-360 of connexin 43, the 'heart' gap junction protein, were employed to immunolocalize connexin 43 gap junctions in the neonatal rat molar tooth germ. Connexin 43 appears early in the differentiation of both ectodermally derived and ectomesenchymally derived cells of the developing tooth. Connexin 43 immunoreactivity is present in the epithelial components of the enamel organ, including the area of the proximal and distal junctional complexes of the ameloblast layer, and the stratum intermedium, stellate reticulum and outer enamel epithelium. Secretory odontoblasts and developing alveolar bone also display a pattern of connexin 43 immunostaining. Both the epithelial and ectomesenchymally-derived components of the developing tooth acquire connexin 43 channels in a manner that correlates with cell differentiation. In addition, three regions can be defined by connexin 43 immunostaining: the epithelia of the enamel organ that are derived from the oral epithelium, the odontoblast layer derived from the ectomesenchyme, and the alveolar bone. The results suggest that connexin 43 may provide the mechanism for functional compartmentalization of the tissues associated with tooth formation. Compartmentalization suggested by connexin 43 expression could play important roles in the development and functions of these tissues.

Differential effect of TGF-beta 1 and PDGF on proliferation of periodontal ligament cells and gingival fibroblasts.

Regeneration of periodontal tissues requires orchestration of several cell types. Two cell types, gingival fibroblastic cells (gingival fibroblasts) and cells from the periodontal ligament (PDL cells), were studied to compare the effects of supplemental addition of TGF-beta 1 and PDGF on proliferation. Cells obtained from healthy donors were cultured in 10% FBS supplemented with either 10 ng/ml TGF-beta 1, 20 ng/ml PDGF, or both. Thymidine incorporation was measured after 24, 48, or 72 hours. Data from PDL (analyzed by ANOVA) showed the following relations: at 24 hours TGF beta 1/PDGF = PDGF > TGF-beta 1 = control; at 48 hours TGF beta 1/PDGF > TGF-beta 1 > PDGF > control; at 72 hours TGF beta 1/PDGF > TGF-beta 1 > PDGF = control. Gingival fibroblast cultures showed the following relations: at 24 and 48 hours TGF beta 1/PDGF = PDGF > TGF-beta 1 = control; at 72 hours, TGF beta 1/PDGF = PDGF > control with TGF beta 1 not different from control or factor combinations. Both TGF-beta 1 and TGF-beta 1/PDGF showed a significantly greater increase in proliferation of PDL cells than in gingival fibroblasts at 48 and 72 hours (Student t test P < 0.05). In contrast, PDGF stimulated proliferation of gingival fibroblasts was significantly greater than PDL cells at 72 hours (P < 0.05). Thus, supplementation of complete cultures (containing 10% FBS) with TGF-beta 1 alone or combined with PDGF stimulates proliferation of PDL cells to a significantly greater extent than proliferation of gingival fibroblasts.(ABSTRACT TRUNCATED AT 250 WORDS)

Continuously infused calcium hydroxide: its influence on hard tissue repair.

To study the ability of calcium hydroxide to promote hard tissue repair, Alza Alzet Osmotic Pumps, implanted in Sprague-Dawley rats, were used to deliver either calcium hydroxide and glycerol, barium hydroxide and glycerol, tetracycline and glycerol, or glycerol only to a standardized round bur defect in a rat femur. The pumps infused one of the reagents into the defects continuously over a 4-wk experimental period. The effects of each reagent on the healing of the bony defects were compared by histological evaluation. The Alza Osmotic Pump proved to be an effective method to deliver an agent to an experimental site. Our preliminary findings from a limited sample size indicated that calcium hydroxide contributed to a more complete osseous repair than either barium hydroxide or tetracycline. Barium hydroxide with a sustained pH equivalent to calcium hydroxide showed no greater healing than the controls. Tetracycline results were also similar to controls.

Identification of the rat bone 60K acidic glycoprotein as alpha 2HS-glycoprotein.

Previous reports have described an Mr 60,000-64,000 glycoprotein present in guanidium chloride (GdmCl)/EDTA extracts of bovine and rat bone. We have purified this protein from the long bones of rats and have raised polyclonal antibodies to the purified protein. The 60K glycoprotein has amino acid and carbohydrate compositions that are similar to those reported for the 60-64K protein(s). Several lines of evidence indicate that the 60K bone glycoprotein is the rat homologue of human alpha 2HS-glycoprotein. First, immunochemical data demonstrated that the 60K bone glycoprotein was present in serum as well as in EDTA/GdmCl extracts of bone. Second, immunolocalization and metabolic labelling experiments showed that the 60K protein is synthesized in liver and not in bone cells, although it is sequestered in vascularized regions of bone matrix. Finally, the NH2-terminal sequence for the rat 60K bone glycoprotein was highly similar to that of the human alpha 2HS-glycoprotein A chain. A surprising finding was that small amounts of contaminating 60K/alpha 2HS-glycoprotein were found in several protein fractions purified by ion-exchange chromatography of bone EDTA/GdmCl extracts. Because this protein was found to be highly immunogenic, the presence of anti-60K antibodies in anti-sera prepared against purified bone proteins should be considered as a potential problem.

An ultrastructural study of human epithelial rests of Malassez maintained in a differentiated state in vitro.

Healthy human periodontal ligaments (PDL), obtained from the extracted teeth (premolars and third molars), were cultivated for 1-35 days, using a multi-purposes culture chamber (MPCC) equipped with various transparent membranes. The resting state of the epithelial rests of Malassez (ERM), similar to their in vivo counterparts, appeared as small islands or strands with scant cytoplasm containing poorly developed organelles. This state was most effectively maintained in MPCC with a cellophane sheet. MPCC with a Sartorius membrane filter permitted proliferation and emigration of ERM. Proliferating ERM were characterized by more profiles of rough endoplasmic reticulum and free ribosomes, new formation of actin-containing microfilaments, less prominent tonofilaments and desmosomes and loss of gap junctions. Most of these ultrastructural changes are manifested in epithelial cells during wound healing. The emigrating ERM from PDL explants, as well as occasional proliferating ERM within explants, consisted of two cell types--outer basal-like cells, as described above, and inner tonofilament-rich prickle-like cells, suggesting a propensity for differentiation of ERM. The results show the possibility of controlling the growth and differentiation of ERM through the MPCC culture environment.