Fibrinopeptide A and fibrinogenfragment B β 15–42 and their relation to the operative trauma and post-operative thromboembolism in neurosurgical patients

Summary In an earlier study on post-operative thromboembolism in neurosurgery the incidence of deep vein thromboses (DVT) diagnosed by the fibrinogen uptake test and phlebography was reduced to the same extent by two different prophylactic methods (low dose heparin or calf muscle stimulation + dextran). However, patients with lower limb paresis due to a brain lesion experienced relatively often a less successful prophylaxis compared to patients with spinal lesions. There are few reports on successful clinical methods for haematological screening of post-operative DVT. The aim of this study was to examine possible haematological indicators for postoperative thromboembolism and secondarily to elucidate whether there exist some special coagulatory or fibrinolytic characteristics in patients who had been operated upon for brain lesions.We have studied two specific coagulatory factors (FPA reflecting thrombin generation and B 15–42 reflecting plasmin activity) in connection with neurosurgical operations. Patients in the above-mentioned study on post-operative DVT operated upon for malignant cerebral tumours or intracranial vascular disease exhibited postoperatively higher values for FPA compared to other neurosurgical diagnoses. B 15–42 was higher in the malignant tumour group and almost significantly higher in the intracranial vascular group (p<0.065). These differences could not be ascribed to the occurrence of DVT.Another 15 patients divided into a minor and a major lesion group were investigated with determination of both parameters pre- and post-operatively. Concerning FPA an increase was noticed post-operatively compared to pre-operatively in the major lesion group. B 15–42 was higher post-operatively in this group compared to the minor lesion group. The results indicate a hyperactivity of the coagulatory system in patients with surgically treated brain parenchyma lesions. These patients are prone to develop post-operative DVT.     

Postoperative changes in hemostasis analyzed by the serial determination of fibrinopeptides and D-dimer

In order to elucidate the postoperative changes in hemostasis, three molecular markers; fibrinopeptide A (FPA), fibrinopeptide Bβ15-42 (Bβ15-42) and D-dimer, were serially determined in 27 gastric resections (group A), 27 hepatic resections (group B) and 4 probe laparotomies (group C). Unexpectedly, a postoperative hypercoagulable state was transient and of a low magnitude, as determined by the obtained value of FPA. On the other hand, a significant fibrinolysis (an elevation of Bβ15-42) was observed immediately after surgery which continued for over 10 days. The early phase of fibrinolysis, up until the 3rd postoperative day, is likely to be primary fibrinolysis, as it was not accompanied by the formation of D-dimer, which results from the digestion of fibrin by plasmin. The late phase, however, is considered to be secondary fibrinolysis, as D-dimer was elevated during this phase. Despite the different surgical procedures, these changes were basically similar between the patients who underwent gastric resections and those who underwent liver resections. The postoperative changes in hemostasis as presented herein may therefore be the general physiological response, at least against abdominal surgery.     

Zur Bedeutung von β-hCG im serum als tumormarker fur das magenkarzinom

Zusammenfassung Einleitung: Nach neueren Untersuchungen ist davon auszugehen, daß bei des Hälfte von Patienten mit einem Magenkarzinom -hCG-positive Zellen im Tumor immunhistochemisch gefunden werden können. Ziel war daher, systematisch zu untersuchen, inwieweit -hCG-immunreaktive Magenkarzinome von einem Anstieg des Serum--hCG begleitet werden and dieses damit als Verlaufsparameter zur Verfügung steht. Methode: Bei 54 Patienten mit einem Magenkarzinom wurde zur immunhistochemischen Darstellung ein gegen -hCG gerichteter monoklonaler Antikörper (Fa. Sigma, 1:100) im APAAP-System verwendet. Die Auswertung wurde nach positiver and negatives Reaktion graduiert. Parallel wurde im Serum des Patienten -hCG präoperativ mit einem Enzymimmunoassay (MEIA, Fa. Abbot) bestimmt. Tumor-stadium, Grading and Tumor-lokalisation werden in die Auswertung mit einbezogen. Ergebnisse: Es wird bestätigt, daß 41% (22 von 54) des Karzinome, unabhängig von ihrer Lokalisation im Magen, eine positive immunhistochemische Reaktion gegen -hCG auslösen. Es zeigte sich in Abhängigkeit vom Tumorstadium eine positive -hCG-Immunreaktivität in 27% (6 von 22) des Tumoren ohne Lymphknoten- and Fernmetastasierung (T_1–4 N₀ M₀), in 54% (7 von 13) des Tumoren mit Lymphknotenaber ohne Fernmetastasen (T_1–4 N₁ M₀) und in 47% (9 von 35) des Tumoren mit Fernmetastasierung. Schlecht differenzierte Tumoren (G3–4) waren zu 42% (15 von 36) und gut differenzierte Tumoren (G_1–2) nur zu 39% (7 von 18) positiv. Aber lediglich bei einer Patientin war der -hCG-Spiegel im Serum erhöht. Zusammenfassung: Immunhistochemisch -hCG-positive Magenkarzinome werden vermehrt bei fortgeschrittenem Tumorstadium und Schlecht differenzierten Karzinomen gefunden. Diese Kar zinome scheinen aber nicht in ausreichender Menge -hCG ins Serum abzugeben, was zu serologisch meßbar erhöh-ten Werten führt. -hCG im Serum kann daher nicht als Prognosefaktor bzw. zur Verlaufskontrolle herangezogen werden. Abzuwarten bleibt, inwieweit die -hCG-Expression von Tumorzellen u. U. Einfluß auf die Propose der Patienten besitzt.

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Significance of -hCG in the serum as a tumour marker for gastric cancer

Introduction: Recent investigations indicate that in 50% of patients with gastric cancer, -hCG-posiitive cells can be found in the tumour by immunohistochemical investigations. The objective of this study was to investigate how often -hCG-immunoreactive gastric carcinomas were accompanied by an elevation in serum -hCG, that could have been used as a course control variable. Methods: In 54 patients with gastric carcinoma a monoclonal antibody directed against -hCG was used for immunohistochemical marking in the APAAP system. The evaluation was graded positive or negative. In parallel, serum -hCG was determined preoperatively using an enzyme immunoassay (MEIA). Tumour stage, grading and tumour locallization were determinants in the evaluation. Results: We found that 41% (22 of 54) of the carcinomas induced a :positive immunohistochemical response to -hCG, regardless of their location in the stomach. In relation to tumour stage, a positive -hCG immunoreactivity was apparent in 27% (6/22) of tumours without lymph node or distant metastases (TI -4N0M0), in 54% (7/13) of tumours with lymph node and without distant metastases (T1–4N1 M0) and in 47% (9/35) of tumours with distant metastases. Poorly differentiated tumours (G3–4) were positive in 42% (15/36) and well-differentiated tumors (G1–2) in 39% (7/18) of cases. In only 1 patient was the -hCG, level in serum elevated, however. Conclusions: -hCG-Positive gastric carcinomas are found more frequently in advanced tumour stages and poorly differentiated carcinomas. These carcinomas, however, seem not to excrete -hCG in sufficient amounts to produce measurable serum values. Therefore, -hCG cannot be used a prognostic factor or for course control. The relevance of -hCG expression of tumour cells to the patients' prognosis remains obscure.

     

Persistent activation of thrombocytes in neurosurgical patients operated for primary brain tumours

Summary A prospective study was designed to investigate whether platelet hyperactivity exists following neurosurgical removal of primary brain tumours. The level of -thromboglobulin (TG), a protein released by platelets during the activation process, was measured in the plasma of 13 consecutive patients prior to surgery (T 1) and on the first (T 2) and seventh (T 3) post-operative days. A significant and sustained increase in TG levels from a baseline of 20.7±1.7 ng/ml (mean ± sem) at T1 to 37.0±5.2 ng/ml (p<0.005) at T 2 and 35.9±3.7 at T 3 (p<0.005) occurred. When patients were grouped according to tumour malignancy, significantly higher TG levels were found in the malignant group at T 2 (51.8±6.3 ng/ml) when compared to the benign group (30.6±6.0 ng/ml) (p=0.025). Postoperative T 3 levels were linearly correlated to T 1 levels (r=0.58, p=0.04).This significant and sustained platelet activation that occurs following brain surgery may be part of the biochemical sequel leading to a hypercoagulable state and thrombo-embolic phenomena (TEP) in these patients.     

β2-microglobulin in Paget's bone disease

On the basis of earlier findings of increased serum ₂-microglobulin concentration in women with postmenopausal osteoporosis, we decided to study serum ₂-microglobulin concentration in other bone diseases. In 28 patients with untreated Paget's bone disease, serum ₂-microglobulin concentration was normal (1.49±0.41 mg/liter versus 1.36±0.21 mg/liter in 42 control subjects, P= ns), a finding that contradicts reports in the literature. We found that serum ₂-microglobulin concentration was related negatively and significantly (r²=–0.154, P=0.0354) with serum total alkaline phosphatase concentration, but not with serum tartrate-resistant acid phosphatase concentration (p =ns). Urinary elimination of ₂-microglobulin was lower in the patients with Paget's disease than in the controls (34±28 versus 120±21 mg/liter, P<0.001). These findings suggest that ₂-microglobulin behaves similarly to osteocalcin (BGP) in Paget's bone disease and that its concentration remains within normal levels perhaps because of the rate of reuptake of ₂-microglobulin in bone neoformation.     

Fibrinolysis profiles and platelet activation after endothelial cell seeding of prosthetic vascular grafts

There is no convincing evidence that endothelial cell seeding of prosthetic grafts in humans confers any of the advantages seen in animals. However, partial endothelial coverage might exert a subtle effect not detectable with indirect end points such as patency or scintigraphy. This study examined seeded cell function by measuring fibrinolytic and platelet activation markers in patients receiving seeded and control prosthetic grafts. Thirty-two patients were randomly assigned to seeded (n=15) and control (n=17) groups. Preoperatively and 3, 6, and 12 months postoperatively, plasma levels of fibrinopeptide A (FPA), B1–42 fragment, cross-linked fibrin degradation products (XL-FDP), thromboxane A₂ (TXA₂), platelet factor 4 (PF4), and -thromboglobulin (ßTG) were measured. Patients with seeded grafts had significantly lower levels of FPA at 6 and 12 months (p <0.05) and a significant overall group effect (p <0.05). These patients also tended to have higher levels of XL-FDP (p <0.1). No other significant differences were seen. The lower rate of conversion of fibrinogen to fibrin and the trend toward increased fibrinolysis seen in seeded grafts may be due to the metabolic effects of viable retained seeded cells. Although comparable platelet activation indicates that endo-thelial coverage remains limited, seeding may exert an antithrombotic influence at the graft surface.Supported in part by W.L. Gore & Associates, Inc., Flagstaff, Ariz.Presented at the Third International Meeting of the British/Swedish Angiology Societies, Lund, Sweden, May 1994.     

Fibrinogen osaka IV: A congenital dysfibrinogenemia found in a patient originally reported in relation to surgery,now defined to have an Aα arginine-16 to histidine substitution

We discovered a congenital heterozygous dysfibrinogen in a patient and reported this case in relation to surgery some time ago (Jpn J Surg (1988) 18:43–46).³ Further studies on the isolated abnormal population of fibrinogen derived from this patient have revealed that fibrinopeptide A was not cleaved by ancrod, a snake venom-derived thrombin-like enzyme, but by thrombin, slowly but completely. The released fibrinopeptide A components, being the A, AY, and AP peptides, were all found to be abnormal, as evidenced by slightly earlier elution positions on high-performance liquid chromatography, compared with the normal counterparts. By analyzing their amino acid sequence, we have identified an arginine to histidine substitution at position 16 of the A chain, the thrombin cleavage site. Utilizing insolubilized abnormal fibrinogen, we confirmed that the polymerization site assigned to the central E domain, the A site, was exposed by thrombin, but not by ancrod. This dysfibrinogen, designated as fibrinogen Osaka IV, is the second abnormal molecule with an A arginine-16 to histidine substitution identified among Japanese families.     

Leptin,soluble leptin receptor,and transforming growth factor-β1 levels in minimal change nephrotic syndrome

Leptin may contribute to renal pathology in some situations by stimulating transforming growth factor-1 (TGF-1) synthesis. The soluble leptin receptor (sOb-R) is a transport protein contributing to binding and activation of circulating leptin. We investigated the interaction between serum and urinary leptin, TGF-1, and serum sOb-R levels in 38 patients with minimal change nephrotic syndrome (MCNS) aged between 6 and 12 years and 10 age- and sex-matched healthy controls (group III). Patients were divided into two groups: group I, proteinuria exceeding >40 mg/m² per hour and group II, patients in remission. Serum leptin levels in group I were significantly lower than those in group II and group III (P=0.011, P=0.007, respectively). There was a negative correlation between serum leptin levels and proteinuria (r=–0.52, P=0.02) as well as between serum leptin and sOb-R levels (r=–0.82, P=0.000) in group I. Urine leptin and sOb-R levels in group I were significantly higher than in group II (P=0.0021, P=0.001, respectively) and group III (P=0.07, P=0.009, respectively). Serum TGF-1 levels in healthy controls (406±424 pg/ml) were significantly lower than those in groups I and II (P=0.004, P=0.000, respectively). However, no significant correlation was found between the serum TGF-1 and leptin levels in MCNS patients. In conclusion, low serum leptin, high serum TGF-1 and sOb-R levels, and elevated urine leptin concentrations were observed at the onset of MCNS. Since long-term proteinuria and leptinuria might be associated with the progression of renal damage, future in vivo and in vitro studies are needed to explain the interaction between these parameters in different types of nephrotic syndrome.     

Methylated β-cyclodextrins are able to improve the nasal absorption of salmon calcitonin

The absorption enhancing effect of methylated -cyclodextrins on the nasal absorption of salmon calcitonin (sCT) was studied in rats and rabbits. The nasal absorption of sCT following administration without additives was low in both species. The absorption in rats could be largely improved by coadministration of cyclodextrins as apparent from the effect on serum calcium concentrations. Trimethyl--cyclodextrin (TMCD), at a concentration of 5% (w/v), was the least potent enhancer. Randomly methylated--cyclodextrin (RMCD) and dimethyl--cyclodextrin (DMCD), all at a concentration of 5% (w/v), were almost equally effective in decreasing serum calcium levels, and the hypocalcemic responses were similar to those of i.v. and s.c. injected sCT. Absorption enhancement was already achieved with 1% DMCD added to the nasal formulations. In rabbits, only the effect of DMCD on the nasal sCT absorption was investigated. A total serum calcium decrement in 4 hours of 9.4±3.9% (mean±SD) was observed following nasal administration of 12.6 IU/kg sCT with 5% DMCD, comparable to that of i.v.-injected sCT. In conclusion, the methylated cyclodextrins DMCD and RMCD are suitable absorption enhancers for nasal sCT administration, which is expected to have a clinical impact on the therapy with calcitonin.     

Transforming Growth Factor-β1 Gene Polymorphisms and Bone Turnover,Bone Mineral Density and Fracture Risk in Southern Chinese Women

Genetic contributions play an important role in determining bone mineral density (BMD) and bone turnover. Transforming growth factor- (TGF-) is abundant in bone and has been implicated as an important regulator of both bone formation and resorption. Several polymorphisms of the TGF-1 gene have recently been suggested to be associated with BMD and susceptibility to osteoporotic spine fractures. To determine the relationship between TGF-1 polymorphisms and BMD in southern Chinese women, three SNPs at C^–1348-T, T²⁹-C, and T^861-20-C of TGF-1 gene were analyzed in 237 postmenopausal southern Chinese women by RFLP and direct sequencing. BMD at the lumbar spine and hip region, biochemical markers of bone turnover, as well as serum levels of TGF-1 were measured. Only the T²⁹-C polymorphism of TGF-1 gene was associated with BMD and fracture risk. The prevalence of fragility fractures was significantly higher in individuals with TC genotype (P < 0.05). Serum alkaline phosphatase and osteocalcin levels as well as urinary N-telopeptide excretion were significantly higher in women with TC than with TT or CC genotypes, and the difference remained significant after adjusting for age and BMI (all P < 0.05). Women with TC genotype had lower BMD at the trochanteric (P < 0.03) and total hip region (P = 0.05). No difference was observed in the serum TGF-1 levels among the three genotypes. In conclusion, an association between T²⁹-C polymorphisms of TGF-1 gene and BMD, bone turnover as well as fragility fractures were demonstrated in postmenopausal southern Chinese women.     

β-Galactosidase as a marker of HSP70 promoter induction in Dunning R3327 prostate carcinoma cells

Hyperthermia is known to improve the response of tumors to radiation or chemotherapeutic treatment when combined in multimodal strategies. The cellular response to hyperthermia is associated with the synthesis of heat shock proteins (HSP). To study the stress response in prostate cancer we have developed a clone of Dunning R3327 rat prostate carcinoma cells stably transfected with a gene construct containing theE. coli -galactosidase gene driven by theDrosophila HSP70 promoter. The measurement of -galactosidase serves as a rapid and semiquantitative assay of HSP70 gene activation. The Dunning cell clone showed evidence of incorporation of the HSP70/-galactosidase construct within the genomic DNA by Southern blot analysis. When compared to mock-transfected control cells, the clone showed minimal baseline -galactosidase activity, which significantly increased following a hyperthermic stress. The time course of -galactosidase elevation following heat stress paralleled the time course of cellular HSP70 elevation by Western blot analysis. These stably transfected Dunning R3327 cells may provide a useful tool to study the effects of hyperthermia, radiation, and chemotherapeutic agents on the cellular stress response and in the establishment of HSP70 as a marker of cellular resistance in the multimodal treatment of prostate cancer.     

Case Report: Intracerebral Hemorrhage After γ-Knife Surgery for Metastatic Brain Tumor

The authors report a case of a 69-year-old man with metastatic brain tumors who died of spontaneous intracerebral hemorrhage 3 days after -knife surgery. He had been suffering from lung cancer with multiple systemic metastasis. Preoperative magnetic resonance images showed two well-defined round lesions with intratumoral hemorrhage in the left frontal and right occipital lobe. There was no bleeding tendency in the hematological examination and the patient was normotensive. -Knife surgery was performed on both lesions in a single session. However, the patient died of massive intracerebral hemorrhage from the left frontal lesion 3 days after the surgery. There have been no previous reports of mortality resulting from spontaneous intracerebral hemorrhage after -knife surgery in metastatic brain tumors documented in the literature. It is likely that the two events, -knife surgery and spontaneous intracerebral hemorrhage, occurred separately and were not associated. However, it is worth noting that there is a possibility of bleeding after -knife surgery, especially in a metastatic brain tumor with preexisting intratumoral hemorrhage as in our case.     

Hypertension following carotid endarterectomy: the role of cerebral renin production

The cause of hypertension in the immediate postoperative period after carotid endarterectomy is unknown. In order to elucidate the etiology of hypertension following carotid endarterectomy, blood samples were drawn intraoperatively from internal jugular vein and external carotid artery prior to and subsequent to carotid endarterectomy in 20 patients. Renin measurement in these samples produced a ratio of internal jugular vein (cerebral) to external carotid artery (systemic). In pre-endarterectomy samples, this cerebral-to-systemic ratio was 1.0 +/- 0.17. However, in the six patients hypertensive postoperatively, this ratio was significantly (p less than 0.02) higher at 1.39 +/- 0.4 than in 14 patients not hypertensive, 0.99 +/- 0.28. Although this ratio in hypertensive patients reverted to 1.12 +/- 0.24 in the postoperative period, the present study suggests a relation between hypertension after carotid endarterectomy and renin production by the brain.     

Gangliocytoma of the pineal body a case report and review of the literature

Summary Ganglion cell tumours are mostly seen in children and young people, but they are extremely rare, accounting for 0.1–0.5% of all brain tumours. It usually occurs in the floor of the third ventricle and the temporal lobe.Recently we have experienced a pineal gangliocytoma, probably the first ever seen in Japan and the fourth case in the world, and have succeeded in a total removal of it.The case concerns a 51-year-old man who sufferend from intermittent blurred vision and headache of 3 years' duration. CT showed, together with severe hydrocephalus, positive contrast medium enhancement and a somewhat irregular but sharply circumscribed high density lesion suggestive of a meningioma. But the brain scintiscan revealed a badly and irregularly demarcated region of warm activity and having little change with time mainly in the pineal region, which was strongly suspicious of gliomas. Hence this scan was thought to be important in diagnosing this tumour. As an operative procedure, biparieto-occipital craniotomy was successfully performed in the sea lion position to remove the tumour totally. Pathological findings indicated a mixture of dispersion and concentration of giant cells possessing prominent nucleoli, abundant chromatin and a prominent nucleus or several nuclei of varying sizes and process-like cell bodies polygonal or irregular in shape. GFAP stain showing no glial fibres and the tumour was thought to be a gangliocytoma.     

Neonates with congenital diaphragmatic hernia have smaller neck veins than other neonates-An alternative route for ECMO cannulation

Background/Purpose: The aim of this study was to compare the size of the right internal jugular vein between neonates with congenital diaphragmatic hernia (CDH) who need extracorporeal membrane oxygenation (ECMO) and non-CDH neonates. The purpose also was to describe a method for cannulation of the right brachiocephalic vein if the internal jugular vein was too small for ECMO cannulation. Methods: Birth weight and size of the right internal jugular vein were compared between CDH and non-CDH neonates subjected to ECMO treatment. A corrected venous size was calculated, estimating the venous size if the patient's birth weight was 3,000 g. Results: The mean birth weight of the CDH patients (3,214 g) was lower than that of the non-CDH patients (3,497 g; P [lt ] .04). The mean venous size of the CDH patients (12.1 Charierre [Ch]) was significantly smaller (P [Lt ] .001) than that of non CDH patients (13.6 Ch). The corrected venous size also was significantly (P [lt ] .001) smaller among CDH patients. Conclusions: Severely affected neonates with CDH have significantly smaller right internal jugular veins than other neonates. If the patient needs ECMO and if the internal jugular vein is too small for cannulation, the right brachiocephalic vein can be successfully cannulated from the neck instead. J Pediatr Surg 37:906-908.     

Transforming growth factor-β pathway is activated in cholecystolithiasis

Background The etiopathogenesis of cholecystolithiasis is not well defined. Primary dysmotility of the organ, due to fibrosis of the gallbladder wall or muscular dysfunction, is suggested as a crucial factor. Transforming growth factor-beta (TGF-) and connective tissue growth factor (CTGF) are involved in several fibrotic disorders and play a critical role in fibrogenesis, thereby changing the physiological function of the organs. In the present study we analyzed the role of TGF- and its downstream target CTGF in patients with cholecystolithiasis.Methods Gallbladders were obtained from 16 individuals undergoing surgery for symptomatic cholecystolithiasis. Normal human gallbladder tissue samples from five individuals without any history of gallbladder disease were obtained through an organ donor transplantation program. Northern blot analysis, in situ hybridization, and immunohistochemistry were used to analyze the expression of TGF-1 and CTGF in the gallbladder tissue samples.Results By northern blot analysis there was an enhanced TGF-1 mRNA expression (eightfold increase; P<0.04) in the cholecystolithiasis tissue samples in comparison with normal controls. There was also a concomitant increase in CTGF (41-fold increase; P<0.01). By in situ hybridization and immunohistochemistry, CTGF mRNA was localized mainly in the mucosa layer, while intensive staining of the smooth muscle cells with TGF-1 and CTGF was observed. In addition, TGF-1 immunoreactivity was also localized in the fibroblasts and inflammatory cells. TGF-1 m-RNA levels showed a significant relationship with the degree of fibrosis in the tissue samples (P<0.04, r=0.5).Conclusion Our data indicate that TGF- and CTGF are involved in ultrastructural tissue changes in patients with cholecystolithiasis. Activation of the TGF- pathway, predominantly in the remaining mucosa and submucosal layer, indicates that extracellular matrix (ECM) synthesis with subsequent gallbladder wall fibrosis is an important step in gallbladder dysfunction in this disorder.     

Transforming growth factor β1-induced cellular heterogeneity in the periosteum of rat parietal bones

Summary We examined the osteogenesis process in transforming growth factor 1 (TGF-1)-treated neonatal and adult rats, aiming to investigate the age difference in the effect of TGF-1 on mesenchymal cell differentiation. Recombinant human (rh) TGF-1 (20 and 200 ng) was injected onto the outer periostea of the right side of the parietal bone of each rat once a day for 1–12 days starting at the age of either 1 day or 12 weeks. On the day after the final injection, the calvaria was excised and evaluated histologically. In the neonates, the 12-day treatment with rhTGF-1 increased the number of osteoprogenitor cells, resulting in intramembranous ossification. In the adult rats, rhTGF-1 induced differentiation of chondrocytes. Cartilage masses were surrounded by mesenchymal cells, which would differentiate into chondrocytes. The cartilage matrix was partially calcified, with chondrocytes buried therein. In the calcified matrix, marrow cavities containing some multinuclear osteoclasts were formed. These findings indicate that rhTGF-1 stimulated the differentiation of mesenchymal cells into chondrocytes and produced the cartilaginous matrix. rhTGF-1 induced intramembranous ossification of the parietal bone in neonatal rats, and it induced enchondral ossification in adults. This result suggests that the different responses of mesenchymal cells in the periosteum to rhTGF-1 may depend on the age of the animals used: namely, they may reflect the respective osteogenic stages of modeling and remodeling.Presented in part at the 4th Workshop on Cells and Cytokines in Bone and Cartilage, Davos, January 11–14, 1992     

Long-term follow-up of a paediatric case of lipoprotein glomerulopathy

A paediatric case of lipoprotein glomerulopathy, a new kidney disease characterized by glomerular lipoprotein thrombi, is reported. The patient had massive proteinuria from the age of 8 years, when the nephrotic syndrome was first detected. This was resistant to conventional treatment for more than 10 years. During the course of the disease, the hyperlipidaemia characteristic of hyper-pre--lipoproteinaemia and elevation of apoprotein E persisted, and renal function gradually deteriorated. The renal histopathological findings from four biopsies were essentially the same, with storage of -lipoprotein in dilated, balloon-like glomerular capillary lumina. However, the number of glomeruli showing global sclerosis increased and tubulo-interstitial changes progressed in parallel with the gradual clinical deterioration. As in other cases reported in Japan some familial involvement has been noted.     

Alendronate reduces serum TNFα and IL-1β, increases neutrophil counts,and improves bone mineral density and bone metabolism indices in patients with chronic idiopathic neutropenia (CIN)-associated osteopenia/osteoporosis

The current study was undertaken to investigate the effect of alendronate on bone mineral density (BMD), bone metabolism markers, and serum bone-resorbing cytokines in patients with chronic idiopathic neutropenia (CIN)-associated osteopenia/osteoporosis. Sixteen randomly selected women, 7 with CIN-associated osteoporosis and 9 with CIN-associated osteopenia, and 14 age- and menopausal status-matched healthy volunteers, were enrolled in the study. Patients received 10mg alendronate daily per os for 360 sdays and studies were done before treatment (day 0) and at varying time points during the study. We found that patients BMD measurements increased by 5.32% after treatment, and that the elevated serum osteocalcin (OC), a bone formation marker, decreased by day 30, normalized by day 90, and increased again by day 270 of treatment. Elevated values of patients urine deoxypyridinoline (Dpd) and N-telopeptide of type I of collagen (NTx), two bone resorption markers, returned to the control range by day 30 and decreased thereafter. Increased levels of patients serum tumor necrosis factor- (TNF) and interleukin-1 (IL-1), two bone resorbing cytokines, returned to the control range by day 30 and decreased thereafter. Peripheral blood neutrophil counts increased by day 30 and continued to rise thereafter, reaching a mean value higher than 2650 neutrophils per µl of blood on day 360. Interestingly, alendronate-induced changes in the levels of both cytokines correlated inversely with the respective changes in neutrophil counts and BMD measurements, and positively with the changes in the respective means of urine NTx and Dpd values. All these findings indicate that alendronate is effective in treating CIN-associated osteopenia/osteoporosis, and that the beneficial effect of the compound may lie, at least in part, in its property to inhibit the production of TNF and IL-1 by cells of the monocyte/macrophage system, in which osteoclasts are included.     

Calciotropic hormones in elderly people with and without hip fracture

The effects of age on calciotropic hormones are not completely understood. The presence of secondary hyperparathyroidism has previously been demonstrated, particularly in patients with hip fracture. The role of a disturbance of vitamin D metabolism, especially a defect in l-hydroxylation, is debated. The aim of this study was to compare serum parathyroid hormone (PTH), osteocalcin and vitamin D metabolites (25(OH)D and 1,25(OH)2D) in osteoporotic elderly patients with hip fracture (HF) and in elderly controls. We studied 57 HF patients aged 83.9±5.9 years (mean±SD) and 68 controls aged 82.5±5 years recruited during two periods: 1 January and 30 April 1988 and 1989. Patients with chronic renal failure (serum creatinine above 150, µmol/l), cancer, or other metabolic bone disease were excluded. Thirty healthy young adults were studied in 1989 only for measurement of 1,25(OH)2D. (1,25(OH)2D was measured by different laboratories in 1988 and 1989 for technical reasons.) We also measured serum PTH, osteocalcin, total calcium and ionized calcium. 1,25(OH)2D levels were not statistically different between HF patients and controls for the two years, nor between HF patients and young healthy adults in 1989. 25(OH)D was decreased in HF patients (p<0.003), as was ionized calcium. Serum PTH levels were higher in HF patients than in controls (p<0.01). A positive correlation has been found between PTH and age in HF patients (r=0.29;p<0.03) and in the whole group of HF patients and controls. There was a significant decrease in osteocalcin in HF patients versus elderly controls (p<0.04). Our results confirm the high levels of intact PTH in elderly HF patients, this elevation of PTH being known to increase bone resporption. Low serum osteocalcin in HF patients seems to reflect decreased bone formation. Thus, this association contributes to the accelerated bone loss in hip fracture. This study also suggests that 1,25(OH)2D is not significantly lowered in case of hip fracture, and l-hydroxylase is not deficient, in spite of a lack of the substrate of this enzyme (25(OH)D). Therefore, a defect of 1,25(OH)2D does not appear to be a pathogenetic factor in bone aging.