[Relationship between marked hyperferritinemia and hemophagocytic lymphohistiocytosis].

OBJECTIVE: To investigate the relationship between marked hyperferritinemia (MHF) and hemophagocytic lymphohistiocytosis(HLH). METHODS: The clinical data of 123 patients with MHF admitted to Peking University People's Hospital from January 2017 to September 2018 were collected, including demographics, baseline characteristics, signs and symptoms, blood routine, blood biochemistry, coagulation function parameters, such as prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib), d-dimer (D-D), fibrin degradation product (FDP), blood ferritin, natural killer (NK) cell activity, soluble interleukin (IL)-2 receptor and bone marrow examination. According to the diagnosis of HLH, the patients were divided into HLH group and non HLH group. The patients were divided into death group and survival group according to the 3-month follow-up results. The groups were compared and statistically analyzed. RESULTS: In the 123 patients with MHF, the average age was (44.2±17.4) years with a male/female ratio of 1.3 ∶1. The most common causes were hematolo-gic malignancies, rheumatologic and inflammatory disorders, iron overload, and HLH. HLH was enriched as the ferritin increased, and the HLH ratios were 28.8%, 40.0%, 54.5%, 50.0%, 50.0% in ferritin value of 10 000-19 999, 20 000-29 999, 30 000-39 999, 40 000-49 999 µg/L, more than 50 000 µg/L respectively. There were 46 cases of HLH, among which 15 cases were secondary to malignancies, 14 cases secondary to rheumatologic disorders, 2 cases secondary to infection, and 15 cases with no clear precipitating cause. There were significant differences between the HLH group and non-HLH group in hepatomegaly, splenomegaly, lymphadenectasis, albumin (ALB), fibrinogen(Fib), P < 0.05, and no significant differences in age, gender, fever, disturbance of consciousness, ferritin level on presentation, maximum ferritin level, cytopenia in 2 or more cell lines, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), direct bilirubin (DBIL), triglyceride (TG), coagulation parameters (PT, APTT, D-D, FDP, exception of Fib), and mortality rate (P > 0.05). There were significant differences between the death group and survival group in disturbance of consciousness, platelet count, PT, TBIL, and DBIL (P < 0.05), but no significant differences in age, gender, fever, hepatomegaly, splenomegaly, lymphadenectasis, ferritin level on presentation, maximum ferritin level, neutrophils, hemoglobin, ALT, AST, ALB, TG, coagulation parameters (Fib, APTT, D-D, FDP, exception of PT) and the HLH ratio (P > 0.05). CONCLUSION: HLH was enriched as the ferritin increased, but marked hyperferritinemia was not specific for HLH in adults.

Diagnostic development for parallel wave-number measurement of lower hybrid waves in EAST.

An eight-channel magnetic probe diagnostic system has been designed and installed adjacent to the 4.6 GHz lower hybrid (LH) grill antenna in the low-field side of the Experimental Advanced Superconducting Tokamak (EAST) in order to study the n∥ evolution of LH waves in the first pass from the launcher to the core plasma. The magnetic probes are separated by 6.6 mm, which allows measurement of the dominant parallel refractive index n∥ up to n∥ = 5 for 4.6 GHz LH waves. The magnetic probes are designed to be sensitive to the magnetic field component perpendicular to the background magnetic field with a slit on the casing that encloses the probe. The intermediate frequency stage, which consists of two mixing stages, down-coverts the frequency of the measured wave signals at 4.6 GHz to 20 MHz. A bench test demonstrates the phase stability of the magnetic probe diagnostic system. By evaluating the phase variation of the measured signals along the background magnetic field, the dominant n∥ of the LH wave in the scrape-off layer has been deduced during the 2019 experimental campaign. In the low density plasma, the measured dominant n∥ of the LH waves is about 2.1, corresponding to the main peak 2.04 of the launched n∥ spectrum. n∥ deduced by the least-squares linear fit method remains near this value in the low density plasma with a high spatial correlation magnitude of 0.9. With an eight-channel probe system, a wave-number spectrum has also been deduced, which has a peak near to the measured dominant n∥.

[A report of six TAFRO syndrome: clinical characteristics, diagnosis and treatment analysis].

Objective: To explore the clinical characteristics, treatment and prognosis of TAFRO syndrome. Methods: All patients diagnosed as Castleman disease in Peking University People's Hospital between December 2011 and April 2019 were included.Among them,6 patients were diagnosed as TAFRO syndrome. Medical records were studied;the clinical manifestation, laboratory test, pathology, treatment and prognosis were analyzed. Recent related literatures were reviewed. Results: The average age of six TAFRO syndrome patients (5 males)was 41.5 years(range, 27-59 years). The patients presented as acute or subacute onset, manifested as fever, thrombocytopenia, polyserositis including pleural effusion and ascites, organomegaly, anasarca, and renal insuffciency. One patient was accompanied by hemophagocyticsyndrome, one patient was accompanied by hypothyroidism, six patients' serum IL-6 was elevated, four patients had received the test of serum VEGF and results were all elevated, six patients' HIV antibody were negative,four patients had received HHV-8 DNA test and results were all negative. For pathology, threewere plasma cell type, twowere mixed type andonewashyaline vascular type. Renal biopsies were performed in 2 patients, showing that renal thrombotic microangiopathyassociated with subacute tubulointerstitial nephritis and secondary capillary proliferative glomerulonephritis. CHOP chemotherapy wereused in 2 patients, glucocorticoid was used in 1 patient, and glucocorticoid combined with Rituximab or Tocilizumab were used in 3 patients. Among them, one patient died because of disease progression after 5 years, other five patientsare still stable. Conclusion: TAFRO syndrome is a rare disease, early recognition and appropriate treatment may improvethe prognosis.

Effectiveness of laser-assisted treatments for medication-related osteonecrosis of the jaw: a systematic review.

The feasibility of laser-assisted treatments of medication-related osteonecrosis of the jaw (MRONJ) remains poorly understood, so we have therefore systematically evaluated their effectiveness. We made a comprehensive search of MEDLINE, Pubmed, and Embase to find randomised controlled trials, case-control studies, and prospective cohort studies that assessed them. We assessed the eligible studies in duplicate, and if possible conducted a meta-analysis. Ten studies with a low to high risk of bias met the inclusion criteria. We found that a comparison of pain scores before and after using visible and infrared GaAs laser in the low-level laser treatment based on the Numerical Pain Rating Scale (mean difference 4.28; 95% CI 3.62 to 4.93; p<0.00001), showed that there were significant differences in the amount of pain. The effectiveness of other laser-assisted treatments on the reduction of pain - for example, Er:YAG laser surgical treatment, and laser-assisted treatment plus platelet-rich plasma, and the effect of other techniques on wound healing of laser-assisted treatments, are uncertain. We found that the results of the studies that were deemed to be high-to-low quality and to have high-to-low statistical power suggested that there may be considerable clinical improvement in MRONJ by using laser-assisted treatment; we cautiously consider that low-level laser treatment may manage pain and symptoms in these patients. More randomised studies of good quality and with a low risk of bias are needed to test whether laser-assisted treatment should be a routine part of management of patients with MRONJ.

Chymotrypsin with sialendoscopy-assisted surgery for the treatment of chronic obstructive parotitis.

Chronic obstructive parotitis (COP) is a common disease of the parotid gland. A total of 104 patients with COP were identified and randomized into a treatment group (52 cases) and a control group (52 cases). All patients underwent sialography and salivary gland scintigraphy (SGS) examinations before surgery. The patients in the treatment group received chymotrypsin combined with gentamicin via interventional sialendoscopy to irrigate the duct, and the control group received gentamicin alone. All patients were asked to record their pain on a visual analogue scale (VAS) before treatment and at 1 week, 2 weeks, 1 month, 3 months, and 6 months after surgery. The VAS score for pain intensity was decreased at 1 week post-treatment in both groups (P<0.05). Compared to the control group, the VAS score was lower in the treatment group at 1 week, 2 weeks, and 1 month post-treatment (P<0.05). The 6-month postoperative SGS results showed improved uptake and excretion in both groups (P<0.05). The treatment group exhibited higher scores for postoperative SGS excretion than the control group (P<0.05). The administration of chymotrypsin combined with gentamicin by sialendoscopy is effective for the treatment of non-stone-related COP and specifically improves the excretion function of the parotid gland.

Scenario analysis of carbon emissions' anti-driving effect on Qingdao's energy structure adjustment with an optimization model, Part II: Energy system planning and management.

In this study, an inexact multistage stochastic mixed-integer programming (IMSMP) method was developed for supporting regional-scale energy system planning (EPS) associated with multiple uncertainties presented as discrete intervals, probability distributions and their combinations. An IMSMP-based energy system planning (IMSMP-ESP) model was formulated for Qingdao to demonstrate its applicability. Solutions which can provide optimal patterns of energy resources generation, conversion, transmission, allocation and facility capacity expansion schemes have been obtained. The results can help local decision makers generate cost-effective energy system management schemes and gain a comprehensive tradeoff between economic objectives and environmental requirements. Moreover, taking the CO2 emissions scenarios mentioned in Part I into consideration, the anti-driving effect of carbon emissions on energy structure adjustment was studied based on the developed model and scenario analysis. Several suggestions can be concluded from the results: (a) to ensure the smooth realization of low-carbon and sustainable development, appropriate price control and fiscal subsidy on high-cost energy resources should be considered by the decision-makers; (b) compared with coal, natural gas utilization should be strongly encouraged in order to insure that Qingdao could reach the carbon discharges peak value in 2020; (c) to guarantee Qingdao's power supply security in the future, the construction of new power plants should be emphasised instead of enhancing the transmission capacity of grid infrastructure.

Collagen fibril stiffening in osteoarthritic cartilage of human beings revealed by atomic force microscopy.

OBJECTIVE: This study aimed to characterize the in-situ mechanical property and morphology of individual collagen fibril in osteoarthritic cartilage using indentation-type atomic force microscopy (IT-AFM). METHODS: The specimens with intact articular cartilage (AC), mild to severe degenerated cartilage from osteoarthritis (OA) were collected with informed consent from the postmenopausal women who underwent hip or knee arthroplasty. The fresh specimens were cryo-sectioned by layers with 50µm thick for each from the articular surface to calcified cartilage, and then processed for AFM imaging and nanoindentation test. For each layer, a total of 20 collagen fibrils were randomly selected for testing. AFM tips with the nominal radius less than 10nm were employed for probing the individual collagen fibril, and the obtained cantilever deflection signal and displacement were recorded for calculating its elastic modulus. RESULTS: An intact AC exhibited a gradation in elastic modulus of collagen fibrils from articular surface (2.65 ± 0.31 GPa) to the cartilage-bone interface (3.70 ± 0.44 GPa). It was noted in mildly degenerated OA cartilage that the coefficient of variation for mechanical properties of collagen fibers, ranging from 25% to 48%, significantly increased as compared with intact one (12%). The stiffened collagen fibrils occurred at either articular surface (3.11 ± 0.91 GPa) or the cartilage-bone interface (5.64 ± 1.10 GPa), accompanied by loosely organized meshwork with advancement of OA cartilage degeneration. It was echoed by histological findings of OA cartilage, including fibrotic changes of surface region and tidemark irregularities. CONCLUSION: The stiffened collagen fibrils in AC occurred with OA onset and progression, not only at articular surface but also the cartilage-bone interface.

A serum-resistant polyamidoamine-based polypeptide dendrimer for gene transfection.

A serum tolerant polycation gene vector, G(2) PAMAM-PGlu-G(1) PAMAMs (ALA), was designed, synthesized, characterized and evaluated. A honeycomb-like molecular structure model for mechanistic explanation of ALA was postulated and discussed. Designed as a star-shaped polyamidoamine (PAMAM)-based polypeptide dendrimer through peptide bond linkages, ALA was with non-toxic low generation G(2) PAMAM (G(2)) as its central core, polyglutamate (PGlu)s as its star-shaped backbone branches and G(1) PAMAM (G(1))s as its branch grafts and peripheral terminals. IR, (1)H NMR demonstrated its successful combination. As a gene carrier, ALA exhibited good DNA binding and condensation capacity with particle size (approximately 87 nm for N/P 40, approximately 170 nm for N/P 30) and ζ-potential (approximately 16 mV for N/P 30-40), negligible cytotoxicity, exciting serum tolerant capacity and significant serum-promoted (serum-containing 56.6%>serum-free 32.7%), cell line dependent (Hek 293 > Bel 7402 > Hela), incubation period dependent (38 h > 18 h > 12 h > 9 h > 4 h > 2 h > 1 h) and sustained (peak transfection appeared at 30 h incubation) transfection efficiency. The presence of serum had not only no inhibition on, but also prominent promotion to, the transfection activity of ALA. All above features differentiated ALA clearly from most other serum-inhibitive nonviral gene carriers, and proved ALA the promising and challenging potential efficient gene vector for practical clinical application.

MMP-3 response to compressive forces in vitro and in vivo.

During orthodontic tooth movement, bone resorption occurs at the compression site. However, the mechanism underlying resorption remains unclear. Applying compressive force to human osteoblast-like cells grown in a 3D collagen gel, we examined gene induction by using microarray and RT-PCR analysis. Among 43 genes exhibiting significant changes, cyclo-oxygenase-2, ornithine decarboxylase, and matrix metalloproteinase-3 (MMP-3) were up-regulated, whereas membrane-bound interleukin-1 receptor accessory protein was down-regulated. The MMP-3 protein increases were further confirmed by Western blot. To ascertain whether MMP-3 is up-regulated in vivo by orthodontic force, we examined human bone samples at the compressive site by realigning the angulated molars. Immunohistochemical staining revealed MMP-3 distributed along the compressive site of the bony region within 3 days of compression. Since MMP-3 participates in degradation of a wide range of extracellular matrix molecules, we propose that MMP-3 plays an important role in bone resorption during orthodontic tooth movement.

[Plant regeneration from protoplast culture in flax (Linum usitatissimum)].

Shoot protoplasts of four fiber flax (Linum usitatissimum) varieties (7309, 948, Belinka and Viking) were isolated and cultured. The optimal condition for higher protoplast yield 1.8 x 10(6)/gFW and activity 85.5% (c.v. 948) were from 10 day old seedings. Culture in V-KM Agroase-island medium led to first divisions after 3 days (c. v. 948), and after twenty days with an efficiency of 36% of divided cells and 5.2% in plating efficiency. Plant regeneration was obtained in 7309 and Belinka on agar media B5-2, MS3 containing 0.6 mg/L 6-BA and 0.1 mg/L NAA. Roots and leaves regeneration were observed in Viking and 948 respectively.

Phosphorylation of the proteins of the extracellular matrix of mineralized tissues by casein kinase-like activity.

The extracellular matrix of the connective tissue contains non-collagenous proteins (NCP) which are acidic in character. The NCP of mineralizing systems (bone, dentin) differ from those of the non-mineralizing systems (skin, tendon) in that the mineralized tissue NCP are frequently phosphorylated. The phosphorylated proteins have been implicated in various aspects of the mineralization process. Thus, it is of interest to consider the mechanism and regulation of phosphorylation of the major matrix NCP. The majority of the phosphorylation takes place at Ser or Thr residues embedded within acidic sequences, and therefore are targets for casein kinase I (CK1) or casein kinase II (CK2)-like kinases. CK1 and CK2 are distantly related members of the protein kinase family. They are ubiquitous, constitutively active, second-messenger-independent kinases. CK1 is found in a variety of isoforms, all homologous to the alpha-subunit of the protein kinase family. It acts as a monomer. The active form of CK2 is a tetrameric holoenzyme, with 2 alpha catalytic subunits and 2 beta regulatory subunits. The CK2 alpha has activity alone, but the holoenzyme is four- to five-fold that activity. CK2 can use either ATP or GTP as the phosphate donor, but CK1 can use only ATP. The CK2 activity which phosphorylates the mineralized tissue NCP appears to be localized to membrane-associated cell fractions, and is present in the endoplasmic reticulum and Golgi compartments in osteoblasts, where phosphorylation of the secreted proteins appears to take place as co- and post-translational processes. Data indicate that both alpha and beta subunits of the membrane-associated CK2 are isoforms of the cytosolic CK2 in the same cells. The CK1 has not been specifically localized. Studies of dephosphorylated NCP such as phosphophoryn (PP) have shown that CK1 will not phosphorylate dephosphorylated dPP unless prior phosphorylation with CK2 has been carried out. In turn, CK2 activity may be initiated only after an initial phosphorylation of one of the messenger-dependent kinases. Thus, the phosphorylation reactions in mineralized tissues may be a tightly regulated hierarchical or sequential cascade of intracellular phosphorylation events.

Characterization and partial purification of microsomal casein kinase II from osteoblast-like cells: an enzyme that phosphorylates osteopontin and phosphophoryn.

Microsomal casein kinase II (mCKII) is a membrane-bound enzyme present in the microsomal fractions of ROS 17/2.8 osteoblast-like cells. It phosphorylates acidic matrix phosphoproteins such as phosphophoryn and osteopontin. Addition of 1.0% Nonidet P-40 facilitates extraction of the optimum amount of detergent-solubilized and -activated enzyme from microsomal fractions. mCKII was partially purified over 3000-fold by sequential chromatography over DEAE-cellulose and heparin-agarose. SDS-polyacrylamide gels, showed that mCKII contained 43 kDa and 31 kDa polypeptides, corresponding to the alpha- and beta-subunits of the enzyme, respectively. The alpha subunit was identified by anti-CKII antiserum and the beta subunit, by its ability to undergo autophosphorylation. The enzyme was inhibited by 50% with 0.4 micrograms/ml heparin and stimulated by 100% with 1.0 mM spermine when casein was used as a substrate. The phosphorylation of phosphophoryn was reduced to 50% by 0.8 micrograms/ml heparin, but was increased to 2-2.5 fold by 5 to 15 mM spermine, which may be due to substrate-directed effects. Kinetic analysis showed that the apparent Km values for phosphophoryn (0.39 microM) and for osteopontin (2.1 microM) were lower than that for casein (21.3 microM). Vmax values of phosphophoryn and osteopontin were 2.2-fold and 4.6-fold higher than that of casein. Using the ratio Vmax/Km as a measure of kinetic specificity, osteopontin and phosphophoryn appear to be the more specific substrates than casein for mCKII. Thus, both proteins can be considered as physiological substrates for mCKII.

Microsomal casein kinase II in endoplasmic reticulum- and Golgi apparatus-rich fractions of ROS 17/2.8 osteoblast-like cells: an enzyme that modifies osteopontin.

Osteopontin is an acidic phosphoprotein containing casein kinase II (CKII) phosphorylatable sites and an acidic amino acid cluster. The metabolically 32P-labelings of both serines and threonines in vitro in osteopontin immunoprecipitated from rat osteoblast-like ROS 17/2.8 cells may suggest that casein kinase II catalyzes this modification. The enzyme occurs in microsomal fractions of rat osteoblast-like ROS 17/2.8 cells. Subcellular fractions containing endoplasmic reticulum and Golgi apparatus were isolated by differential centrifugation and were identified according to their ultrastructures and the presence of marker enzymes such as glucose-6-phosphatase and thiamine pyrophosphatase, respectively. both fractions phosphorylated the partially dephosphorylated osteopontin and the specific substrate peptide RRREEETEEE. Endoplasmic reticulum-catalyzed peptide phosphorylation was 2.7 times lower than that of Golgi although both endoplasmic reticulum- and Golgi-catalyzed peptide reactions were 50% inhibited by 20 and 100 ng/ml heparin, respectively. Western blot analysis revealed that both fractions contained osteopontin and microsomal CKII. Furthermore, microsomal CKII was immunogold-labeled in endoplasmic reticulum and Golgi apparatus. Heparin inhibition and utilization of [gamma-32P]GTP as a phosphate donor by both fractions confirmed their capacity to phosphorylate osteopontin. The results suggest that microsomal CKII modifies the acidic matrix proteins during transportation. These matrix phosphoproteins may participate in the mineralization process of hard tissues.

Autosomal dominant, familial spastic paraplegia, type I: clinical and genetic analysis of a large North American family.

"Familial spastic paraplegia" (FSP) refers to clinically and genetically diverse syndromes characterized by insidiously progressive lower extremity spasticity. We evaluated 126 members of a large kindred, including 31 affected subjects, in which FSP was transmitted as a stereotyped, autosomal dominant disorder that showed complete genetic penetrance. Affected subjects developed insidiously progressive gait disturbance between ages 12 and thirty-five. Neurologic examination revealed hyperreflexia and spasticity in the lower extremities, weakness of hip flexion and ankle dorsiflexion, extensor plantar response, diminished vibratory sense in the feet, and pes cavus. Using genetic linkage analysis, we excluded the FSP1 locus on chromosome 14q11.2 as the disease locus in this family. We present the clinical and genetic features of FSP type I, including the age-adjusted risk of developing the disorder in this family.

[Studies on magnetic responsiveness and renal embolization with magnetic microspheres in dogs].

The responsiveness and transcatheter embolization with magnetic gelatin microspheres (MG-ms) in dog kidney were reported. In experiments with magnet using a constant flow apparatus such as roller pump, the percent retention was determined by counting the carrier retained by the magnetic field and divided by the total starting counts. Factors influencing the percent retention of MG-ms include: velocity of medium flow, magnetic field intensity, alpha-Fe2O3 content in the MG-ms, viscosity of the medium and so on. Transcatheter embolization with MG-ms (10-30 microns) was performed under external magnet control in dog kidney. The result of angiogram and histological section showed that the MG-ms were in the arteries, arterioles, and glomerular capillaries with no adverse reactions. The embolized effect with magnet field was more prominent than the group without magnet field. These results show that the MG-ms is a promising embolic agent for treatment of renal cancer under external magnet control.

[Studies on distribution of magnetic gelatin microspheres in rabbits].

In this report, the technique of labelling MG-ms with 99mTc as pertechnetate in the presence of a reducing agent such as SnCl2 was described. The distribution of intravenously injected 99mTc-labelled MG-ms in rabbits at different intervals of magnetic field applied and different magnetic field intensity was investigated by using an externally applied magnetic field and measuring the radioactivity at the rabbit head and other organs. When magnet was used, the radioactivity in the head, target site, was 15 times more than that when magnet was not used. At the same time, the radioactivity of the lung was 5 times less than when magnet was not used. The newly designed magnetic field equipment was presented.

[Studies on pharmacologic disposition of adriamycin in dog after hepatic arterial embolization with adriamycin carboxymethyl-dextran-microspheres].

The ion exchange adriamycin carboxymethyl-dextran-microspheres (AD-CM-DMS) was chosen as a model preparation. Pharmacokinetics, targeting and embolization effects of the microspheres were studies after hepatic arterial embolization in vivo in dogs. After hepatic arterial infusion, the concentrations of ADM in peripheral vein blood and hepatic tissue were determined by HPLC. The peak concentrations were (0.558 micrograms/ml for AD-CM-DMS and 1.013 micrograms/ml for ADM solution. T1/2 (alpha), T1/2 (beta) and MRT of AD-CM-DMS were respectively 2.82, 3.19 and 1.28 times as much as those of ADM solution. At the target site, the tissue concentrations of AD-CM-DMS were respectively 8.0 and 9.1 times those of ADM solution. The dynamic vessel angiography revealed no external and internal collaterals and no complete degradation of AD-CM-DMS after sixteen weeks was observed. These results suggest that AD-CM-DMS are useful as embolic agent for the treatment of hepatic cancer. It could facilitate intensive chemotherapy with minimum systematic side effect.

The in vitro phosphorylation of the native rat incisor dentin phosphophoryns.

Phosphophoryns are the major non-collagenous proteins of the mineralized matrix of rat incisor dentin. Nearly half the phosphophoryn residues are serines, and 85-90% of these are phosphorylated. Since phosphorylation may be important for phosphophoryn function, it was of interest to identify the kinase(s) responsible for catalyzing their phosphophorylation. Rat osteosarcoma (ROS) 17/2.8 osteoblast-like cells were selected as the enzyme source. Native rat incisor phosphophoryns (RIPP-I, II, III) were not substrates for any of the ROS 17/2.8 messenger-dependent kinases but were phosphorylated by membrane-associated endogenous messenger-independent kinases. These were resolved chromatographically and identified as casein kinase (CK) I and II by elution properties and immunoblotting with a CKII antibody. The CKI preferentially used RIPP-III as substrate, while CKII preferred RIPP-I and II. Heparin at 100 and 500 ng/assay and NaCl at 0.25-0.4 M inhibited phosphorylation of the RIPP by CKI and CKII in parallel. At 10 mM spermine, phosphorylation of RIPP-I and II by CKII, and of RIPP-III by CKI were inhibited, but phosphorylation of RIPP-III by CKII was enhanced. Purified sea star oocyte CKII demonstrated the same substrate specificity and spermine concentration shift as the ROS 17/2.8 CKII. These data show that osteoblast-like cells are a rich source of membrane-bound CKI and CKII activity. The different patterns of phosphorylation of RIPP-I, II, and III further show that they are distinct synthetic products of the odontoblast.

Phosphorylation of extracellular bone and dentine matrix proteins.

Phosphoproteins appear to be involved in several ways in the regulation of the orderly deposition and crystal growth of mineral within the performed collagenous matrix of bone and dentine. The phosphorylation of these proteins is not yet understood. Potential protein kinases were extracted from an osteoblast-like cell line, ROS 17/2.8. The ROS 17/2.8 line was shown to produce a full complement of known kinases. However, neither bone phosphoproteins (BPP) nor dentine phosphophoryn (DPP) could be phosphorylated by the messenger dependent kinases. DPP and dephosphorylated BPP (dBPP) were substrates for a unique messenger independent kinase distinct from casein kinase II, and dDPP was a still better substrate. Thus, BPP and DPP are phosphorylated by a unique kinase or set of kinases which are messenger independent and have very specific substrate sequence requirements.