Sulfur compounds in therapy: radiation-protective agents, amphetamines, and mucopolysaccharide sulfation.

Sulfur-containing compounds have been used in the search for whole-body radiation-protective compounds, in the design of amphetamine derivatives that retain appetite-suppressive effects but lack most behavioral effects characteristic of amphetamines, and in the search for the cause of kidney stone formation in recurrently stoneforming patients. Organic synthetic procedures were used to prepare radiation-protective compounds having a variety of sulfur-containing functional groups, and to prepare amphetamine derivatives having electron-attracting sulfur functions. In the case of the kidney stone causation research, isolation of urinary mucopolysaccharides (MPS) from recurrently stoneforming patients was carried out and the extent of sulfation of the MPS was determined by electrophoresis. Whole-body radiation-protective agents with a high degree of protection against lethal doses of gamma-radiation in mice were found in a series of quinolinium and pyridinium bis(methylthio) and methylthio amino derivatives. Mechanism studies showed that the copper complexes of these agents mimicked the beneficial action of superoxide dismutase. Electron-attracting sulfur-containing functions on amphetamine nitrogen, as well as 4'-amino nitrogen provided amphetamine derivatives with good appetite-suppressant effects and few or no adverse behavioral effects. Higher than normal levels of sulfation of the urinary MPS of stone formers suggested a cause for recurrent kidney stone formation. A sulfation inhibitor was found to prevent recurrence of stone formation and inhibit growth of existing stones. The inclusion of various sulfur-containing functions in organic molecules yielded compounds having whole-body radiation protection from lethal doses of gamma-radiation in animals. The presence of electron-attracting sulfur functions in amphetamine gave derivatives that retained appetite-suppressant effects and eliminated most adverse behavioral effects. A therapy for recurrent urolithiasis resulted from inhibition of MPS sulfation, after the finding that stoneforming patients had abnormally high levels of MPS sulfation.


Critical analysis of the content and enforcement of mandatory consultation and patient profile laws.

This study evaluates the experience of 12 states that mandate that pharmacists provide consult services to patients and maintain drug profiles. An analysis of each state's statutes and regulations was complemented by telephone interviews of state licensing board staff. Nine states specify the information to be provided during consultations, but only one (Florida) requires a comprehensive list of drug information, including adverse drug reactions, drug-drug interactions, directions for use, and necessary warnings. Eleven of the 12 states also require pharmacists to maintain patient drug profiles, but only 6 states specify the type of information to be included in profiles. State efforts to inform consumers about the pharmacist consultation requirement are limited, with only 7 states undertaking minimal promotional programs. The fact that no states reported any consumer consultation complaints or disciplinary actions against pharmacists for failure to consult illustrates the limited nature of mandatory consultation statues and regulations. The wide variety of requirements demonstrates that these laws do not represent a uniform standard of practice. The lack of documented evidence regarding implementation invites questions regarding their usefulness and reinforces the need for enhanced monitoring activities.


[Review of results and clinical analysis of conservative partial surgery associated with postoperative radiotherapy in operable breast cancers stages I amd II].

Over the last 25 years, conservative forms of treatment have been used for the locoregional treatment of early breast cancer, stages I and II (< 3 cm in diameter). At present, the same carcinological results are obtained with conservative surgery and radiation therapy as with radical surgery, and the aesthetic and functional results have improved. Several parameters should be taken into account, ie tumour volume, breast volume, multifocal nature and histologic type. A number of questions still remain unanswered namely: what is the most appropriate conservative surgery (quadrantectomy, tumourectomy or local excision)? What is the most appropriate total irradiation dose (50 Gy over 5 weeks: or 45 Gy over 4.5 weeks)? What is the value of radiation boost in patients with negative resection margins? What is the value of radiation boost in patients with a high local risk of recurrence? What is the most appropriate radiation technique for boost in the primary tumour bed (electrons or iridium implants)? Does the external irradiation of regional lymph nodes improve survival rate? Is it possible to extend conservative treatment to a 4-cm diameter tumour? What is the impact of adjuvant systemic chemotherapy and/or hormonotherapy on the risk of isolated breast cancer recurrence?


E-cadherin expression: a counterbalance for cancer cell invasion.

Invasion, eventually leading to metastasis, is presented as the result of a balance between the activation of 2 sets of genes, coined i+ (invasion promotor) and i- (invasion suppressor) genes. Experiments in vitro have indicated that the homotypic homophilic epithelial cell--cell adhesion molecule E-cadherin (L-CAM; uvomorulin; cell CAM 120/80; Arc-1; rrl antigen) is an i- gene product. In several cell families, manipulation of E-cadherin at the level of the protein by antibody-mediated inactivation, at the level of the mRNA by antisense DNA transfection, and at the level of the genome by sense DNA transfection respectively resulted in induction and suppression of invasiveness. Nude mouse tumors from non-invasive homogeneously E-cadherin-positive cell populations were found to be invasive and metastatic. These tumors expressed E-cadherin in a heterogeneous manner, the undifferentiated cells being negative; but tumor-derived cells in culture were again E-cadherin-positive, indicating downregulation of this protein by host factors. Several types of human cancers showed a similar heterogeneity suggesting a relationship between downregulation of E-cadherin and invasion. Our current research focus is on the factors responsible for E-cadherin downregulation in experimental and human cancers.


[Regulation by protein kinase C of TGF-beta 1 expression in cultured cells of breast adenocarcinoma].

In human breast carcinoma MCF-7 cells, phorbol diesters inhibit proliferation and induce cell maturation. We investigated the involvement of TGF-beta 1 in the PCK-mediated inhibition of breast cancer cell proliferation. Using an RNase protection assay, we showed that TPA induced a dose-dependent increase in levels of TGF-beta 1 mRNA that paralleled the inhibitory effect on MCF-7 proliferation. Similar results were obtained with another TPA-sensitive breast cancer cell line (BT-20). TPA did not increase TGF-beta 1 mRNA levels in the MCF-7:RPh-4 and T47D cell lines, which are both insensitive to the growth inhibitory effects of phorbol esters. In addition, the increase in TGF-beta 1 mRNA level was not observed after treatment of the MCF-7 cell with other inducers of cell differentiation such as forskolin, DMF, HMBA and sodium butyrate. The induction of TGF-beta 1 mRNA by TPA along with its inhibitory effect on cell proliferation suggests that TGF-beta 1 mediates, at least in part, the inhibitory effect of PKC activation.


[Apoptosis or programmed cell death: concepts, mechanisms and contribution in oncology].

Programmed cell death, or apoptosis, corresponds to a sequence of intracellular events that lead to cell death. It has been shown that apoptosis is necessary in some physiological conditions such as embryogenesis, homeostasis of the immune system, erythropoiesis, etc. Some xenobiotics can induce apoptosis at lower doses and necrosis at higher doses. When a cell dies, it is either by apoptosis or by necrosis, and there are many differences between these two deaths. Apoptosis begins by a pre-commitment phase, which is reversible; during this phase the cell has a high level of second messengers. The commitment phase then follows and is irreversible, even when the xenobiotic that triggered the induction is removed. Most often, apoptotic cell death requires synthesis of macromolecules, the inhibition of their synthesis can prevent it. The cell undergoes important morphological changes during apoptosis, its volume decreases when its density increases. Then chromatin becomes granular, intensively osmiophilic, it condenses along the nuclear membrane. Later, chromatin disintegrates into small granules which will be phagocytized. One of the most important characteristics of the programmed cell death is the activation of an endonuclease, that gives rise to DNA fragments of 180-200 base pairs or multiples of these numbers; then after electrophoresis, the DNA gives the appearance of a ladder. Apoptotic cells can be characterized after classic staining, and flow cytometry; they can be separated from other cells by centrifugation on a gradient of density. It has been hypothesized that cell transformation could be due to a sudden resistance to apoptosis. However, the most interesting aspect in oncology recently demonstrated is that well-known anticancer drugs are able to induce apoptosis. One can hope that the discovery of new targets for anticancer drugs could lead to discovering new drugs that could be more active.


[Growth factors and intestinal cancers].

This review focuses on the growth factors (primarily IGFs, TGF-alpha and TGB-beta) that control both proliferation and differentiation of the normal intestinal epithelial cells and their involvement in intestinal tumorigenesis. Integrity of the digestive tissue is dependent on continuous coordination between cell growth and maturation along the crypt- villus axis. Beyond an intricate network of various regulatory molecules, such a regulation is essentially in close connection with the opposite biological effects delivered on a same target cell by TGF-alpha and TGF-beta. Growth factors act via regulatory autocrine/paracrine loops that are physiological means to deliver biological signals throughout the normal gut tissue. During tumorigenesis, cell progressively lose their sensitivity towards such extracellular regulatory loops. TGF-alpha insensitivity is linked to constitutive activation of intracellular pathways that induce uncontrolled cell growth. The incapacity to respond to TGF-beta that is due to an alteration of its intracellular pathway does not allow the negative regulation of cell proliferation or the induction of cell differentiation. Concurrently, the disappearance of an IGF-II extracellular autocrine loop appears to be correlated with cells maintained in an undifferentiated state. These alterations lead to a break between the metabolic pathways involved in the delicate control of the proliferation/differentiation balance. This leads to an unscheduled increase of positive proliferative signals which are responsible for an uncoordinated epithelial cell growth that favour tumor cell clone outgrowth. From these experimental data, essentially obtained in vitro, we propose a tentative colorectal tumorigenesis model that links both growth factor pathways and genetic (oncogenes and tumor suppressor genes) alterations. However, such a model only represents a part of the multiple cell and molecular interactions that are set in action in vivo. It remains to decipher their consistency in order to improve new therapeutic strategies.


[Mammary microcalcifications: is there a consensus on interpretation and management?].

Twenty two view mammograms without magnification, with microcalcifications corresponding to non palpable breast lesions, were submitted to 23 experienced observers, either gynaecologists or radiologists. These mammograms consisted of 10 malignant lesions and 10 benign lesions; all of them underwent surgical procedures. The microcalcifications were graded and categorized into one of the four groups: malignant; suspect; slightly suspect; benign. One of the four options was proposed: biopsy; mammogram within 3 months; mammogram within 6 months; mammogram within a year. Twelve observers out of 23 have referred to a classification. The mammograms were assessed according to a consensus. The validity of that diagnostic test was studied in various clinical situations and the lack of homogeneity of the responses was quantified. In the trade-off situation between sensitivity and specificity (suspect or malignant considered as positive), sensitivity is 50% and specificity is 70%. Moreover, the responses are not homogeneous at all and this lack of homogeneity is found to be statistically significant, greater than by chance alone. The diagnosis of cancer is more frequently put forward by the radiologists. This study explains the low predictive value of non palpable microcalcifications undergoing surgical procedures: from 11.5 to 44% in the literature. Similarly, there is no agreement on the frequency of the follow-up for these microcalcifications when they do not lead to surgical procedures.


[Meta-analysis of adjuvant medical treatment in breast cancers. Ten year results].

The ten-year results of the meta-analysis of the adjuvant systemic treatment of early breast cancer are summarized. The five-year results are confirmed and a few new informations arise: tamoxifen is more efficient when given more than two years; the results of chemotherapy are significant in post-menopausal patients less than 70 years old.


[Puncture of bone lesions under tomodensitometric control].

Sixty CT-guided fine needle biopsies were performed on 59 patients. Of the 40 lesions in patients with a previous history of cancer, malignancy was confirmed by cytological examination in 27 cases and benignity in three cases. Of the 20 lesions in patients with no previous history of cancer, malignancy was confirmed in nine cases (45%) and benignity in five cases (25%). This precise and safe procedure shows 78% overall accuracy in this series.


A phase II trial of early intensive chemotherapy with autologous bone marrow transplantation in the treatment of poor prognosis non seminomatous germ cell tumors.

Twenty-eight patients with poor prognosis, advanced metastatic non seminomatous germ cell tumors (NSGCT) were treated with early high dose chemotherapy and autologous bone marrow transplantation (ABMT) rescue. The primary tumor was testicular in 19 patients and extragonadal in nine patients. For 19 patients with a testicular primary, the median probability of complete remission (CR) was 0.05 according to our prognostic mathematical model based on pretreatment levels of serum HCG and AFP. The same prognostic model was used for extragonadal primaries. Treatment consisted of two cycles of a modified double dose of cisplatin, vinblastine, bleomycin, VP-16 regimen (mPVeBV) followed by a high dose cisplatin-etoposide-cyclophosphamide regimen (PEC) followed by ABMT. Of the 28 patients, 17 (61%) achieved CR, one of which was surgical CR (sCR), five died of rapidly progressive disease early during the first cycle of mPVeBV, two had treatment-related deaths, three did not respond and one patient refused treatment. Of the 17 patients initially in CR, three relapsed after 4, 4 and 7 months respectively and have subsequently died. Two other patients died while still in CR: one committed suicide and one died of an infectious complication due to transfusion-related AIDS. Twelve patients are alive in CR after a median follow-up of 66 months (range 7-72 months). The non parametric 3-year survival rate is 40%. To demonstrate the effect of intensive chemotherapy with ABMT, a randomized multicenter French study was set up to evaluate the PVeBV regimen with or without high dose treatment and ABMT in poor risk NSGCT patients.


Histocompatibility antigens associated with Behçet's disease in northern Han Chinese.

Among Han nationality Chinese and living in the northern area of the Yellow River, 120 patients suffering from Behçet's disease and 100 unrelated healthy individuals were typed for histocompatibility antigens (HLA)-A, -B, -C, and -DR and -DQ antigens. HLA-DR and DQ typing was performed on B-lymphocyte separated with Lympho-B-Kwik. The HLA-antisera were provided by 11th IHWC. Bf alleles and C4 allotypes were determined by immunofixation agarose-gel electrophoresis. HLA-B51 was found in 67/120 (55.83%) patients and in 12/100 (12%) controls, the Chi-square and relative risk values were 45.54 and 9.27, respectively (p < 0.0005). C4AQ0 frequency was significantly increased in the patient group. In the complete form group HLA-B51 was observed more frequently (62.79%). No significant differences of other HLA antigens, frequencies, Bf or B4 alleles were found between the groups.


Type 1 (insulin-dependent) diabetes mellitus with coexisting autoimmune thyroid disease in Japan.

Type 1 diabetes mellitus is known to be a heterogenous disease which is frequently complicated with other autoimmune thyroid diseases (AITD). The present study was designed to investigate the clinical characteristics and HLA antigens in Japanese Type 1 diabetic patients with AITD. Subjects were 25 Type 1 diabetic patients with AITD (13 Graves' disease and 12 Hashimoto's thyroiditis) and 32 Type 1 diabetic patients without AITD. Compared with Type 1 diabetic patients without AITD, age at onset of diabetes was later and positive ICA persisted much longer in the diabetic patients with AITD. Compared with normal controls, DR9 was increased in the patients with AITD, while DR4 was increased in those without AITD. Type 1 diabetic patients with AITD were characterized by the late onset of diabetes, persistent ICA and increased association with DR9. These results suggest that immunological and genetic heterogeneity may exist within Japanese Type 1 diabetic patients.


Central motor conductivity in aged people.

The conductivity of motor neurons in 26 aged females (mean age 79 years) was analyzed by the conventional conduction method and by pulsed magnetic stimulation and compared with that in 14 younger controls. In aged people, slow motor conduction velocities were found in peripheral nerves. Central motor conduction time (CMCT) in relaxed muscle was shorter in the aged people, although CMCT was normal in mildly contracted muscle. These findings coincide with the results studied in Parkinson's disease, although these subjects were not diagnosed as having Parkinson's disease. Aged people generally have an anteflexed posture, slow movements, and poor postural reflexes, and have been reported to have a decrease in the dopamine level which is relatively earlier than that of other transmitters in the basal ganglia. This may account for the present finding that aged people have neurophysiological abnormalities in CNS which are similar to those in Parkinson's disease.


Unoperated thoracic aortic aneurysms: survival rates of the patients and determinants of prognosis.

The purpose of this study was to evaluate the long-term prognosis of unoperated thoracic aortic aneurysms, and to detect the risk factors which determine the prognosis of unoperated patients. The subjects were 52 unoperated patients with 58 thoracic aortic aneurysms (22 of the ascending aorta, 36 of the descending aorta or the aortic arch) and 38 with abdominal aortic aneurysms. The survival rates of the patients with ascending aortic aneurysms at 3 years and 5 years and those of the patients with descending aortic aneurysms at 5 years were significantly higher than those of the patients with abdominal aortic aneurysms. The risk factors for rupture of thoracic aortic aneurysms were the large size of aneurysms and non-management by the cardiologist and the risk factors for death unrelated to the aneurysms were patient age, male gender and non-management by the cardiologist.


Phonoechocardiographic identification of chordal snap in hypertrophic cardiomyopathy.

In three patients with hypertrophic cardiomyopathy, a late systolic click following mid systolic murmur was recognized. M-mode and two-dimensional echocardiography showed the presence of systolic anterior motion of the chordae tendineae in all cases. Phonoechocardiographic studies showed that the time of the click always coincided with the endpoint of mild anterior motion of the chordae tendineae. The click was considered to be a chordal snap.


Endobronchial involvement with Waldenström's macroglobulinemia.

Although many cases of Waldenström's macroglobulinemia (WMG) with pulmonary involvement have been reported, endobronchial involvement has not been described. We report an unusual case of WMG wherein endobronchial invasion of tumor cells narrowed the airways and caused coughing and wheezing.


Pentasomy X mosaic in two adult sisters with diabetes mellitus.

Pentasomy X mosaic in two adult sisters with non-insulin dependent diabetes mellitus is described. The younger sister had schizophrenia, and both were mentally retarded, but no apparent somatic abnormalities were found. Chromosome analyses revealed karyotype 45,X/46,XX/47,XXX/48,XXXX/49,XXXXX mosaic with a low frequency of aneuploidy on cultured peripheral lymphocytes and 46,XX on cultured skin fibroblasts in both sisters. The low frequency of X chromosome aberration may be responsible for the lack of somatic abnormalities and the long life in both sisters. The association of pentasomy X mosaicism and diabetes mellitus however appears to be coincidental.


Anaplastic thyroid carcinoma with prominent cardiac metastasis, accompanied by a marked leukocytosis with a neutrophilia and high GM-CSF level in serum.

Cardiac metastasis of thyroid carcinoma is extremely rare. We treated a case of anaplastic thyroid carcinoma with prominent cardiac metastasis. The 61-year-old male was admitted because of high fever. Investigations revealed a cardiac mass and anaplastic thyroid carcinoma. Resection of the cardiac mass revealed that it was metastasis from the thyroid carcinoma. After 4 months, he died in spite of intensive therapy. Marked leukocytosis was observed during the clinical course, and a concomitant increase of granulocyte macrophage-colony stimulating factor (GM-CSF) level was demonstrated in the sera. It was suggested that the high GM-CSF level in serum contributed to leukocytosis.


Partial anomalous pulmonary venous return showing anomalous venous return to the azygos vein.

Partial anomalous pulmonary venous return (PAPVR) is a congenital heart disease with a reported incidence of autopsied case. The location of the anomalous pulmonary venous return is usually the right atrium, superior vena cava (SVC), and sometimes the brachiocephalic vein, inferior vena cava (IVC) or coronary venous sinus. Recently we experienced a rare case of PAPVR showing anomalous right total pulmonary venous return to the azygos vein. Furthermore, downward translocation of the right upper lobe bronchus was evident. This rare case is reported along with a review of the related literature.


Thyroid papillary adenocarcinoma in a woman with Peutz-Jeghers syndrome.

We present a rare case of Peutz-Jeghers(P-J) syndrome in a 29-year-old woman who developed adenocarcinoma of the thyroid and polyps in the gastrointestinal tract. Polyps removed from the stomach, colon and ileum showed features typical of hamartomas. Histological examination indicated that the tumor resected from the right lobe of the thyroid was a papillary adenocarcinoma. This appears to be the first precise report of a P-J syndrome associated with thyroid carcinoma. This case adds an example of predisposition of P-J syndrome to extra-intestinal malignancy and indicates the importance of long-term observation of malignancy.


Bartter's syndrome with gouty arthritis.

A 45-year-old woman with Bartter's syndrome complicated with gouty arthritis is reported. Bartter's syndrome was diagnosed from hypokalemia, metabolic alkalosis, high plasma renin activity, normal blood pressure and attenuated pressor response to exogenous angiotensin II infusion. Serum uric acid was 11.8 mg/dl, the renal clearance of uric acid was low (3.0 ml/min) and fractional excretion of uric acid (FEUA) was 7.0%, being the lower limit of the normal range. Fractional excretion of phosphate was low (5.3%) and fractional free water clearance per fractional distal sodium delivery was 0.51. The mechanism of hyperuricemia in this case is discussed.


Multiple liver abscesses secondary to Yersinia enterocolitica.

A 37-year-old male, a poorly-controlled insulin-dependent diabetic patient, was admitted to our hospital with complaints of high fever and confusion. Laboratory data showed hyperglycemia, positive inflammatory reaction and liver dysfunction. Blood culture demonstrated Yersinia enterocolitica. Liver CT scan showed multiple low density areas. These data were consistent with a diagnosis of liver abscess secondary to Yersinia enterocolitica. He died of disseminated intravascular coagulation; subsequent autopsy confirmed the clinical diagnosis. Liver abscess secondary to Yersinia enterocolitica with septicemia is rare, but has been reported in compromised hosts. In the mechanism of this disease, the alimentary tract has been suggested to be the port of entry in most cases.


Wegener's granulomatosis in a patient with a rheumatoid arthritis.

A 38-year-old woman with rheumatoid arthritis who developed Wegener's granulomatosis is described. Wegener's granulomatosis appeared with saddle nose, perforation in her nasal septum, and granuloma in the nasal cavity. Laboratory evaluation showed a positive rheumatoid factor and circulating immune complex. Radiographic examination revealed ankylotic changes in both wrist and elbow joints. Bilateral anosmia and other disease manifestations completely responded to treatment with oral cyclophosphamide and prednisolone.


Immunological studies on opportunistic infection and the development of adult T-cell leukemia.

Three patients with opportunistic infection preceding adult T-cell leukemia (ATL) are presented: a 66-year-old woman with cryptococcosis and Pneumocystis carinii pneumonia, a 46-year-old man with Pneumocystis carinii pneumonia, and a 55-year-old woman with cryptococcosis. Although, at the first examination, the first two had smoldering type ATL and the third case was an HTLV-I carrier, all three developed overt ATL 14-16 months after the onset of opportunistic infection. It is considered that there is immune suppression already present in HTLV-I carriers and in patients with smoldering ATL, and opportunistic infection is predictive of the development of ATL.


Post-splenectomy meningitis in patients with lymphoid malignancy.

This report describes pneumococcal meningitis in two splenectomized patients with lymphoid malignancy. Early diagnosis and emergency treatment of pneumococcal infection in splenectomized patients is proven to be of significant value. Prophylactic antibiotics targeted against pneumococcus should be considered for the clinical management of such conditions in splenectomized patients, particularly during cytotoxic chemotherapy.


Idiopathic acute eosinophilic pneumonia.

A previously healthy young man presented with acute respiratory distress, high fever and bilateral ground-glass appearance on chest radiograph. Bronchoalveolar lavage analysis demonstrated significant eosinophilia (72%) with no evidence of infection. The transbronchial lung biopsy showed that the walls of bronchioli and alveolar septa were markedly infiltrated with eosinophils. The patient rapidly improved with corticosteroid therapy. This case exemplifies the recently described idiopathic acute eosinophilic pneumonia. Similar cases published in the Japanese literature were reviewed and discussed.


Persistent hemichorea associated with thyrotoxicosis.

We describe a case with unilateral chorea associated with thyrotoxicosis. A 23-year-old female with no family history of neurological diseases acutely developed choreic movements of the left extremities during gross thyrotoxicosis. CT scan and MRI study demonstrated no abnormality. Single-photon emission CT with technetium Tc 99m-labeled hexamethylpropyleneamine oxime revealed normal cerebral perfusion. Although the choreic movements were partially improved by dopamine antagonist, they persisted for two months until successful treatment of the thyrotoxicosis finally abolished these movements. Increased sensitivity of dopamine receptors may be responsible for persistent choreic movements in thyrotoxicosis.


Granulomatous hypophysitis with meningitis and hypopituitarism.

We report an unusual case of granulomatous hypophysitis in which visual impairment, meningitis and hypopituitarism in a 76-year-old female were associated with radiological evidence of a pituitary mass. The sellar lesion was indistinguishable from pituitary tumor on neuroimaging studies, but the recovery of visual acuity and visual field abnormalities together with the improvement of pituitary function after steroid administration indicated that the mass lesion was due to an inflammatory disease of the pituitary gland. The pituitary tissue obtained by transsphenoidal hypophysectomy revealed granulomatous inflammatory cell infiltration with epithelioid cells and scattered multinucleated giant cells. Although a causal relationship with meningitis was not ascertained, possible exposure of the CSF space to the autoimmune inflammatory process of the pituitary gland was likely in view of the positive pituitary antibody reaction and radiological evidence of suprasellar extension. This entity should be considered when evaluating patients with a pituitary mass, hypopituitarism and meningitis.


Thoracic extramedullary hematopoiesis associated with hereditary spherocytosis.

A 54-year-old man was diagnosed with hereditary spherocytosis. An X-ray of the chest disclosed bilateral round masses in the upper and lower posterior mediastinum. CT showed homogeneous round masses of soft tissue density in the posterior mediastinum. A needle biopsy of the mass showed hyperplastic erythroid bone marrow. Fatty metamorphosis of the masses was observed in CT 2 years after splenectomy but showed no change in size.


Bronchopulmonary disease in ulcerative colitis.

Two cases of ulcerative colitis are described: a 33-year-old woman who developed widespread bronchiectasis 7 months after undergoing colectomy, and a 72-year-old man whose colonic disease began coincidentally with the appearance of diffuse interstitial pulmonary infiltrates. In both cases, clinical correlation and common patterns of response of lung and bowel diseases suggested that the co-existence of these two pathologies might not be merely a casual relation.


Detoxification of nonopiate drugs in the chronic pain setting and clonidine opiate detoxification.

Although the pain physician is most familiar with the treatment of the opiate withdrawal syndrome, other drugs are abused by the chronic pain patient. The pain physician should then be familiar with the withdrawal syndromes associated with other drug groups. The withdrawal syndromes associated with hypnosedatives, psychotomimetics, nicotine, stimulants, ergot alkaloids, beta adrenergic blocking agents, antidepressants, muscle relaxants, and alpha-adrenergic agonists are described. Drug detoxification protocols for these drugs are reviewed. Additionally, the rationale for clonidine opiate detoxification is discussed, and current clonidine detoxification protocols are reviewed.


Chronic-pain medications: equivalence levels and method of quantifying usage.

Medication use is an important consideration in chronic-pain rehabilitation programs (CPRPs). However, it is difficult to quantify this aspect of chronic-pain treatment, because patients often take multiple medications that can differ by pharmacological class as well as dosage level. The Medication Quantification Scale (MQS) provides a method for quantifying medication use in patients with chronic, nonmalignant pain. Scores are calculated for each medication based on weights assigned by medication class and dosage level, and these scores are summed to provide a quantitative index of total medication usage suitable for statistical analysis. The method for calculating MQS scores is illustrated, and research data on MQS reliability and validity are presented. Interrater reliability was rho = 0.985 (p less than 0.0001) for 30 MQS scores calculated by two clinicians. MQS scores for 88 patients correlated well with the clinical judgment of 12 health care professionals (mean rho = 0.755, p less than 0.0001). The MQS scores for 60 chronic-pain patients (30 treated in a CPRP and 30 untreated) were obtained at two time points: evaluation and 1-year follow-up. A two (groups) by two (time points) analysis of variance yielded a significant group-by-time interaction (F = 8.82, p less than 0.0043). Treated patients decreased their medication intake significantly (p less than 0.0001), whereas untreated patients did not (p greater than 0.57). The MQS offers a reliable and valid method for quantifying medication usage in chronic-pain patients.


Patient controlled analgesia: the relation of psychological factors to pain and analgesic use in adolescents with postoperative pain.

To examine the relationship of psychological variables to pain and patient-controlled analgesia (PCA) use in adolescents undergoing orthopedic surgeries. Cohort. Fifty adolescents and their parents. Visual analogue scales of pain, the McGill Pain Questionnaire, and questionnaires that assessed overall psychological distress, anxiety, and family environment. Correlations of the psychological variables with measures of pain and PCA use showed that high levels of anxiety and distress in adolescents were related to higher postoperative pain. Adolescents from controlling and conflicted families also had higher pain ratings and made more frequent PCA demands during the lockout interval when drug was unavailable. Parent anxiety and distress were also related to adolescent pain and PCA use. The psychological status of adolescents and their parents can significantly influence postoperative pain and PCA use.


MMPI scale 3 as a predictor of back injury report: what does it tell us?

Items and selected subscales of Scale 3 (Hysteria) of the MMPI were examined to pinpoint personality or emotional factors predictive of back injury reports in an industrial setting. Data were derived from a previous prospective-design study of back pain in volunteer hourly wage employees of an aircraft manufacturing company. After physical examination and completion of questionnaires pertaining to demographic, psychosocial (including the MMPI), and workplace factors, workers were followed for an average of 3 years. Those who subsequently reported back injury were compared with those who did not. In that study three variables predicted report of back injury, one of which was Scale 3 of the MMPI. Individual items, Ornduff et al. subscales of Psychological Denial and Body Concern, and the five Harris-Lingoes (1955) subscales of Scale 3 were analyzed. Three Harris-Lingoes subscales showed significant relationships to the criterion. Hy-3: Lassitude/Malaise; Hy-1: Denial of Social Anxiety; and, marginally, Hy-2: Need for Affection, significantly contributed to prediction effectiveness. Results and implications for the understanding of factors predicting back injury reports and for the medical evaluation of pain and the concept of pain are discussed.


An 18-month follow-up of a secondary prevention program for back pain: help and hindrance factors related to outcome maintenance.

Thirty-six subjects participated in an 18-month follow-up of a secondary prevention program for back pain of recent onset. The follow-up assessment included most of the same instruments used in the original study. In addition, interviews were conducted to ascertain help and hindrance factors for maintaining improvements during the follow-up period. Results showed that subjects had significantly less pain, used fewer medications, and were more active at 18-month follow-up than at baseline. All subjects had returned to work, and one third had no pain-related work absences during the follow-up. A cost-benefit analysis indicated substantial economic savings when follow-up sick-listing data were compared with estimates based on an increasing trend for pain-related absenteeism found during the baseline period. Hindrance factors reported by subjects were related to personal time-management and workplace factors, especially psychosocial aspects of the work environment. It was concluded that the secondary prevention program was effective and that future maintenance programs should focus more on personal time-management and workplace factors.


Persistent low-back pain is real. However, diagnostic spinal injections are not helpful in its evaluation.

We endeavored to assess the short-term effects of intrathecal fentanyl and lidocaine in chronic-pain patients by ascertaining whether the opioid fentanyl, by virtue of its lack of sensory and motor paralysis, conferred any diagnostic advantages over lidocaine, a local anesthetic whose effects include sensory and motor paralysis. Neuraxial administration of fentanyl has been touted as an improved diagnostic tool to distinguish between peripheral and central pain, because the absence of sensory and motor effects may avert the patient's presumption of the onset of analgesia based on these cues. Twenty-two patients with persistent low-back pain, whose investigations had determined that they were not surgical candidates, were studied using a counter-balanced, placebo-controlled, and double-blinded crossover design. Each patient received three separate lumbar intrathecal injections of equal volume (1.4 ml): cerebrospinal fluid, fentanyl 25 micrograms, and lidocaine 70 mg. Pain and symptom assessments were performed preinjection (baseline), and at regular intervals up to and including 4 h postinjection. Pain was evaluated by verbal patient response using a numerical pain-rating system of 0 to 10. Duration of analgesia, sensation of warmth, and adverse effects were noted. Statistical analyses were performed using nonparametric tests. Subjects' average age was 56 years, with a median low-back pain duration of 16 years. There were no significant differences in the baseline median-pain scores among injection types. The baseline and best cerebrospinal fluid-pain scores were significantly different, suggesting a placebo effect. The best pain scores for fentanyl and lidocaine were superior to their own baseline levels and to the best cerebrospinal fluid scores.(ABSTRACT TRUNCATED AT 250 WORDS)


Chronic back pain: view from a psychiatrist's office.

Twenty-six adult outpatients suffering from chronic back pain were observed in a clinical study. Treatment was multifaceted. A psychiatrist with a subspecialty in pain management served as the coordinator of all treatments used. Treatment included biofeedback, physical therapy, behavioral-management program and pain measurement scales, psychotherapy with pain counseling, and medication. Patients were seen weekly for 4 months and observed for 1 year thereafter. Results of the study bore out the necessity of a coordinated multifaceted approach to treatment of chronic pain. Strong emotional overlay was seen in all patients. Stress, depression, and secondary gain are symptoms that were addressed along with the physical symptomatology. Patients improved significantly. The psychiatric component serving as the anchor for treatment was viewed as a key to success. The emotional component present in chronic pain must not be overlooked, because it plays a vital role in successful treatment.


Voices from the past: ancient views of pain in childhood.

Our understanding of pain in childhood has made many important advances in the past 15-20 years. Examination of the views of ancient writers, who were concerned about pain and disease in childhood, reveals similar concerns that have dominated the discussions of clinicians and researchers in the twentieth century. The most recent pediatric pain research confirms the position of many writers in previous centuries who believed children and infants are sensitive to pain. Although childhood historians have assumed that pain in children was largely ignored in previous centuries, examination of ancient medical writings reveals ongoing debates about the sources of pain in childhood, symptoms of pain and its treatment. This paper contrasts historical perspectives with the views of twentieth century writers.


A case of opiate-insensitive pain: malignant treatment of benign pain.

We report the case of a woman with presumed cancer pain treated with escalating doses of opiates despite no evident improvement in her pain and several deleterious side effects. A 62-year-old woman with cervical myelopathy and a diagnosis of a spinal cord tumor was referred to the University of Washington Medical Center complaining of chest tightness, multiple joint pains, nausea, constipation, seizures and a deteriorating memory. At the time of admission she was confined to her bed with a full-time attendant and was receiving 240 milligrams of intravenous morphine per hour for her pain. Diagnostic studies failed to find any evidence of neoplasm and revealed only an old hemorrhage within the cervical spinal cord. A program of increasing physical and occupational therapy and decreasing opiate intake was initiated. Within a month the patient's pain complaints decreased, as did the rest of her presenting complaints. Her activities of daily living greatly increased making attendant care no longer necessary. This case report illustrates some of the hazards of opioid therapy in the management of patients with chronic pain. Our patient's opiate therapy was expensive, gave her undesirable side effects, and did not reduce her pain complaints or improve her function. In the treatment of chronic pain, of noncancerous or cancerous origin, a) systemic opioids may not be effective in reducing pain complaints in every patient, b) treatment efficacy evaluation should always include functional endpoints, and c) nonefficacious treatments should not be continued indefinitely.


RSD score: criteria for the diagnosis of reflex sympathetic dystrophy and causalgia.

No standardized criteria for the diagnosis of reflex sympathetic dystrophy (RSD) are in common use. An RSD score scoring system of diagnostic criteria is proposed. Diagnosis of reflex sympathetic dystrophy by RSD score correlated well with clinical diagnosis in a group of 25 patients who had treatment for chronic upper extremity pain. Standardization of diagnostic criteria with the RSD score should enhance comparison of the outcomes of studies concerning the treatment of RSD.


Interscalene blocks for chronic upper extremity pain.

A study of 25 patients was carried out to determine the efficacy of interscalene block (ISB) for the treatment of chronic upper extremity pain. An RSD score was used to categorize these patients. Seventeen of the 25 patients had less pain after ISB, and 14 also had increased range of motion of the affected limb. Patients with reflex sympathetic dystrophy (RSD)/causalgia, as well as other chronic pain conditions, improved. ISB was compared with stellate ganglion block (SGB) in patients undergoing both treatments. ISB seemed to be at least as effective as SGB for treatment of RSD/causalgia and may have some advantages over SGB. The role of somatic and sympathetic blockade is discussed.


Reflex sympathetic dystrophy of the stump in below-knee amputees.

To document the occurrence of reflex sympathetic dystrophy of the stump in two patients with below-knee amputation. A retrospective survey emphasising two clinical case reports. Department of orthopaedic rehabilitation at a teaching rehabilitation hospital. Lower limb amputees (n = 164) were accepted for prosthetic rehabilitation. Twenty-one amputees were regarded as rehabilitation failures; in two below-knee amputees intractable pain was the major problem. Clinical manifestations, radiological, and scintigraphic findings in the two amputees with intractable pain met the criteria for diagnosis of reflex sympathetic dystrophy. Reflex sympathetic dystrophy of the stump should be suspected in below-knee amputees whenever severe pain persists over a period of 3 to 4 months following amputation.


Physical illness and the family emotional system: psoriasis as a model.

Most physical illnesses are characterized by significant variability in age of onset and severity of clinical course. Although many diseases may occur in the context of a strong family history for the disease, they may also occur when there is no such history. Psoriasis has been chosen as a model for demonstrating that variability in age of onset, clinical course, and family history for a given disease may be explained, in part, by factors related to the family emotional system, a concept described by family systems theory (Bowen theory). Links between the clinical manifestations of psoriasis and the family emotional system have important treatment implications.


Coping with inner feelings and stress: heavy alcohol use in the context of alexithymia.

The authors conducted a population study to assess the relationship between heavy alcohol consumption and alexithymia, defined as difficulty in identifying and expressing feelings. The study sample consisted of 2,297 middle-aged men from eastern Finland. The proportion of men who reported either frequent intoxication or unpleasant aftereffects of heavy drinking increased linearly with alexithymia. Alexithymia and the heavy acute intake of different sorts of alcoholic drinks were also consistently associated. Long-term heavy use, taking into account both the dose and frequency, was also linearly related to the alexithymia measure. In multivariate models to assess whether high alcohol consumption in alexithymic individuals might relate to stress, the authors found nothing to support the stress-dampening hypothesis.


Psychosocial antecedents of athletic injury: the effects of life stress and social support on female collegiate gymnasts.

The article presents information from two studies of a research program investigating psychosocial antecedents of athletic injury. Study 1 describes the development of the Life Events Survey for Collegiate Athletes (LESCA). The LESCA demonstrated good content validity and provided a stable measure of life stress. The LESCA's validity and the moderating effects of social support were explored in the second study. The LESCA provided a construct-valid measure of life stress and was a better predictor of athletic injury than the Social and Athletic Readjustment Rating Scale. Social support moderated the stress-injury relationship in such a way that LESCA negative life stress accounted for 11% to 22% of injury variances in low-support conditions; LESCA positive life stress accounted for 14% to 20% of injury variances in high-support conditions. Mechanisms underlying the stress-injury relationship and directions for future research are discussed with respect to the model outlined by M. B. Andersen and J. M. Williams in 1988.


Mammalian glycosyltransferases: genomic organization and protein structure.

In recent years, several glycosyltransferase genes and cDNAs have been cloned and characterized. Although the glycosyltransferases seem to share the same general architecture, there is only little sequence similarity between the various enzymes. Moreover, a comparison of the organization of the genes shows that there is no common pattern of intron-exon structure. In addition, there seems to be little or no correlation between glycosyltransferase exons and protein domains. Taken together, these observations suggest that many of the glycosyltransferase genes evolved independently. So far, only two glycosyltransferase gene families have been described. These families may have evolved by exon-shuffling, or by gene duplication and subsequent divergence. For specific glycosyltransferases, mechanisms such as alternative splicing and alternative promoter usage play a role in the production of multiple protein isoforms from a single gene. These isoenzymes may differ in their enzymatic properties or cellular localization.


Photoaffinity labelling of glycosyltransferases.

The photoaffinity analogues 5-azido-UDP-glucose and 5-azido-UDP-glucuronic acid have proven to be valuable biochemical tools in the studies of nucleoside diphosphate sugar-utilizing enzymes, especially membrane-associated glycosyltransferases. A summary of the past and current uses of these analogues is presented, as well as photoaffinity data for the enzyme UDP-glucose: dolichylphosphate glucosyltransferase (Glc-P-Dol synthase). This enzyme has served as a model membrane-associated glycosyltransferase for demonstrating the uses of 5-azido-UDP-glucose. The advantages of using photoaffinity analogues for the purification and characterization of glycosyltransferases are presented, as well as an outline of the general procedures which can be used in conjunction with these analogues.


Identification and synthesis of a novel 15 kDa beta-galactoside-binding lectin in human leukocytes.

Knowledge of the identity, synthesis and secretion of beta-galactoside-binding lectins by leukocytes is of importance because lactosaminoglycans present at the leukocyte cell surface may be physiologically significant lectin receptors that could mediate autocrine or paracrine functions and/or cell adhesion. This paper presents data that show that a previously identified 15.5-16.5 kDa lactose-binding protein synthesized in vitro by human peripheral leukocytes is actually comprised of three different polypeptides. One of these is related to a novel 15 kDa lectin isolated from human spleen and which is synthesized by B lymphoblastoid cells. Spleen contains at least six lactose-binding polypeptides for which the carbohydrate-binding activity is independent of the presence of divalent cations and mercaptoethanol. The splenic 15 kDa polypeptide does not appear to be immunologically related to previously characterized beta-galactoside-binding lectins. It is separable from galaptin, another galactoside-binding lectin (subunit mol. wt 14.5 kDa) by chromatography on DEAE-Sephacel. Western blot analyses and immunoprecipitation/fluorography experiments with metabolically labelled cells showed the presence of the 15 kDa lectin in peripheral leukocytes and in Epstein-Barr virus-immortalized B lymphoblastoid cells. The 15 kDa lectin yielded polypeptide fragments of approximately 6.2 and approximately 8.6 kDa after cyanogen bromide (CNBr) degradation. These fragments were partially sequenced and 12 residues/fragment were identified. A similarity search of the SWISS PROT protein data base did not reveal a relationship of the 15 kDa polypeptide to known lectins, including galaptin.(ABSTRACT TRUNCATED AT 250 WORDS)


Reducing the overlap problem in the proton NMR spectra of oligosaccharides by application of pseudo-four-dimensional homonuclear HOHAHA-HOHAHA-COSY.

A new homonuclear NMR experiment is described for the assignment of the proton NMR spectra of oligosaccharides, namely HOHAHA-HOHAHA-COSY (where HOHAHA is homonuclear Hartmann-Hahn spectroscopy and COSY is correlated spectroscopy). While this technique may formally be thought of as a four-dimensional NMR experiment, by use of selective pulses it is demonstrated that the analogous pseudo-four-dimensional experiment is a valuable improvement over conventional three-dimensional HOHAHA-COSY, in that the degree of resonance overlap is markedly reduced by the dispersion of resonances into a fourth effective dimension. The technique is demonstrated by application to the biantennary nonasaccharide Gal beta 1-4-GlcNAc beta 1-2Man alpha 1-6(Gal beta 1-4GlcNAc beta 1-2Man alpha 1-3)Man-beta 1-4GlcNAc beta 1-4GlcNAc.


Carbohydrate structure of Marburg virus glycoprotein.

Marburg virus was propagated in E6 cells, a cloned cell line of Vero cells, in the presence of [6-3H]glucosamine. Radiolabelled viral glycoprotein was digested with trypsin, and oligosaccharides were liberated by sequential treatment with endo-beta-N-acetylglucosaminidase H, peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F and O-glycosidase, by beta-elimination, and by alkaline hydrolysis. After fractionation by HPLC and gel filtration, glycans were characterized chromatographically, by digestion with exoglycosidases and, in part, by methylation analysis and liquid secondary ion mass spectrometry. The oligosaccharide structures thus established include oligomannosidic and hybrid-type N-glycans, as well as neutral fucosylated bi-, tri- and tetraantennary species, most of which carry an additional bisecting N-acetylglucosamine. In addition, high amounts of neutral mucin-type O-glycans with type-1 and type-2 core structures were detected. None of the glycans present in this viral glycoprotein carried sialic acid residues.


The tertiary structure of endo-beta-1,4-glucanase B (CenB), a multidomain cellulase from the bacterium Cellulomonas fimi.

Endo-beta-1,4-glucanase B (CenB) is a large (110 kDa) extracellular enzyme from the cellulolytic bacterium Cellulomonas fimi. CenB contains five domains, including a typical C.fimi cellulose-binding domain, separated by distinctive linker polypeptides (Meinke et al., 1991b). X-ray scattering analyses show that CenB has a highly elongated shape resembling beads on a string. The sizes of the polypeptides produced by treatment of CenB with proteases, together with their N-terminal amino acid sequences, show that at least two of the four linkers connecting the five domains of CenB are more sensitive to proteolysis than the domains themselves. It is concluded that the beads represent the domains of CenB, the string represents the linkers.


A human lysosomal alpha(1----6)-mannosidase active on the branched trimannosyl core of complex glycans.

Normal human fibroblasts and fibroblasts from a patient with alpha-mannosidosis were grown in the presence or absence of 100 microM swainsonine for 7 days. Accumulated oligosaccharides were isolated and analysed by high performance liquid chromatography (HPLC) and methylation analysis. Man alpha 1----3Man beta 1----4GlcNAc and Man alpha 1----2Man alpha 1----3-Man beta 1----4GlcNAc (where Man is D-mannose and GlcNAc is N-acetyl-D-glucosamine) comprised greater than 80% of the total oligosaccharides in untreated mannosidosis cells. However, Man alpha 1----6[Man alpha 1----3]Man beta 1----4GlcNAc was the major Man3GlcNAc isomer present after 7 days of swainsonine treatment. No mannose-containing oligosaccharides were detected in control fibroblasts in the absence of swainsonine but, in its presence, oligosaccharides containing 2-9 mannose residues accumulated. Man alpha 1----6[Man alpha 1----3]-Man alpha 1----6[Man alpha 1----3]Man beta 1----4GlcNAc and Man alpha 1----6-[Man alpha 1----3]Man beta 1----4GlcNAc were the major components (67%). Surprisingly, Man alpha 1----3Man beta 1----4GlcNAc was only observed in swainsonine-treated control cells during the recovery period after removal of swainsonine. These studies suggest the presence of a second lysosomal alpha-mannosidase activity which is unaffected in genetic alpha-mannosidosis, but is inhibited by swainsonine. This enzyme would cleave the alpha(1----6)-linked mannose residue from branched Man3GlcNAc to form Man alpha 1----3Man beta 1----4GlcNAc. To confirm this hypothesis, fractions from alpha-mannosidosis and control fibroblasts that bound to concanavalin A (ConA)-Sepharose and were eluted with 0.5 M alpha-methyl mannoside were incubated at pH 4.0 with Man alpha 1----6[Man alpha 1----3]Man beta 1----4-GlcNAc. As anticipated, Man alpha 1----3Man beta 1----4GlcNAc was the sole product using enzyme from mannosidosis fibroblasts, while the major product from control fibroblasts was Man alpha 1----6Man beta 1----4GlcNAc. This confirmed the presence of a swainsonine-inhibitable alpha(1----6)-mannosidase activity unaffected by the disease. The differing substrate specificities of the alpha(1----6)-mannosidase and the major lysosomal alpha-mannosidase indicate that the alpha(1----6)-mannosidase plays an important role in the generation of the oligosaccharides accumulated in alpha-mannosidosis patients.


Developmental sialic acid modifications in rat organs.

The changes in expression of sialic acids in Sprague-Dawley rats in the prenatal and early postnatal time period have been examined in multiple organs, both visceral and non-visceral. In all organs examined, there is a dramatic increase in both N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) shortly after birth. The bulk of the sialic acid is present in the ganglioside fraction in all tissues examined. As total amounts of sialic acid present in gangliosides decrease, the proportion present in the low molecular weight cytosolic fraction increases. A curious observation is that Neu5Ac hydroxylase activity is present at the time of the increase in sialic acid, but its activity does not correlate with Neu5Gc expression after the early postnatal period. This implies that Neu5Gc expression has another level of regulation besides CMP-Neu5Ac hydroxylase activity.


Presence of O-linked oligosaccharide on a threonine residue in the human transferrin receptor.

We have previously demonstrated that the human transferrin receptor (TfR) of approximately 90 kDa contains Ser/Thr-linked (O-linked) oligosaccharides. In the present study, we report our identification of the site of attachment of the O-linked oligosaccharides in the receptor. A 70 kDa fragment from the external domain of the TfR was generated by trypsin treatment of the [3H]glucosamine-labelled receptor purified from human K562 cells. The beta-elimination of the intact TfR, but not the 70 kDa fragment, released Gal-[3H]Gal-NAcitol, indicating that the 70 kDa fragment lacks O-linked oligosaccharides. In the remaining 20 kDa fragment there are three potential sites (Thr96, Thr104 and Ser106) for O-glycosylation in the extracellular domain. To identify which of these residues are O-glycosylated, both the [3H]Thr- and [3H]Ser-labelled TfR were directly treated with mild base to effect beta-elimination, and the radiolabelled amino acids and their derivatives were analysed. Approximately 2% of the total radiolabelled Thr, but no radiolabelled Ser, was converted to expected beta-elimination products by this treatment. These and other results demonstrate that only one O-linked oligosaccharide is present in the TfR and that it occurs on either Thr96 or Thr104. From human serum we purified the cleaved, soluble form of the TfR (s-TfR), which contains Thr104, but lacks Thr96. The s-TfR was sensitive to O-glycanase and bound to Jacalin lectin, indicating that the s-TfR contains an O-linked oligosaccharide.(ABSTRACT TRUNCATED AT 250 WORDS)


Identification of the O-linked glycosylation site of the human transferrin receptor.

The human transferrin receptor is a glycoprotein containing three N-linked and one O-linked glycosylation sites. Tryptic digestion of the receptor, followed by chromatography on BioGel P-2 and reverse-phase HPLC, yields a glycopeptide (amino acids 101-120) containing the O-linked site. Amino acid sequence analysis reveals that the site of O-glycosylation is Thr-104. Mass spectral analysis is consistent with the presence of a Gal-GalNAc core with predominantly two sialic acid residues.


Topology of ER processing alpha-mannosidase of Saccharomyces cerevisiae.

The yeast specific alpha-mannosidase which converts Man9GlcNAc to a single isomer of Man8GlcNAc is involved in N-linked oligosaccharide processing in the endoplasmic reticulum (ER). Sequence analysis of the structural gene for this enzyme suggested that it is a type II transmembrane protein (Camirand et al., 1991). To firmly establish its membrane topology, the gene was transcribed in vitro and translation was performed in a reticulocyte lysate with and without dog pancreas microsomal membranes. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of [35S]methionine-labelled products showed that the largest band formed corresponded in size to the 63 kDa peptide expected from the alpha-mannosidase gene product. It was transformed into a 4 kDa larger endoglycosidase H-sensitive band in the presence of microsomal membranes. This glycosylated translation product was completely protected from proteinase K digestion in the absence of detergent. These results demonstrate that the yeast ER alpha-mannosidase is a type II membrane protein, like Golgi enzymes involved in N-linked glycosylation.


Cloning and expression of the murine gene and chromosomal location of the human gene encoding N-acetylglucosaminyltransferase I.

A mouse cDNA clone previously isolated from an F9 teratocarcinoma cell library and shown to confer N-acetylglucosaminyltransferase I (GlcNAc-TI) activity on Lec1 Chinese hamster ovary (CHO) cell transfectants [Kumar, R., Yang,J., Larsen,R.D. and Stanley,P. (1990) Proc. Natl. Acad Sci. USA, 87, 9948-9952] has been sequenced. The nucleotide and deduced amino acid sequences are highly homologous to previously described human and rabbit GlcNAc-TI cDNAs. A 1250 bp portion of the mouse cDNA encoding all but the first 34 amino acids of the deduced protein sequence was inducibly expressed in Escherichia coli and gave rise to a prominent fusion protein of mol. wt approximately 45 kDa whose presence correlated with high levels of GlcNAc-TI activity in cell lysates. Probes generated from the cDNA were used to show that the GlcNAc-TI gene is present in a single copy in mammals and that a homologous gene was not detectable (under low-stringency hybridization conditions) in DNA from yeast, sea urchin, Drosophila or Chaenorhaditis elegans. Genomic DNA clones that hybridized to probes generated from the GlcNAc-TI cDNA were isolated from a mouse liver library. Restriction analyses, Southern hybridization and DNA sequence analyses of subcloned genomic DNA fragments and a polymerase chain reaction (PCR) product provided evidence that the coding and 3' untranslated regions of the cDNA reside in a single exon. However, the mouse GlcNAc-TI gene (Mgat-1) includes at least one additional exon 5' of the coding region. Southern analyses of DNA from mouse-human somatic cell hybrids and in situ hybridization were used to locate the human GlcNAc-TI gene (MGAT-1) between positions q31.2 and q31.3 on chromosome 5, a region of chromosome 5 that is syntenic with a region of mouse chromosome 11. Northern analyses of adult mouse tissues revealed two GlcNAc-TI gene transcripts that are differentially expressed in different tissues.


Fingerprinting genomes by use of PCR with primers that encode protein motifs or contain sequences that regulate gene expression.

PCR primers of arbitrary nucleotide sequence have identified DNA polymorphisms useful for genetic mapping in a large variety of organisms. Although technically very powerful, the use of arbitrary primers for genome mapping has the disadvantage of characterizing DNA sequences of unknown function. Thus, there is no reason to anticipate that DNA fragments amplified by use of arbitrary primers will be enriched for either transcribed or promoter sequences that may be conserved in evolution. For these reasons, we modified the arbitrarily primed PCR method by using oligonucleotide primers derived from conserved promoter elements and protein motifs. Twenty-nine of these primers were tested individually and in pairwise combinations for their ability to amplify genomic DNA from a variety of species including various inbred strains of laboratory mice and Mus spretus. Using recombinant inbred strains of mice, we determined the chromosomal location of 27 polymorphic fragments in the mouse genome. The results demonstrated that motif sequence-tagged PCR products are reliable markers for mapping the mouse genome and that motif primers can also be used for genomic fingerprinting of many divergent species.


Modulation of polymorphic loci detection with synthetic tandem repeat variants.

The modifications of hybridization patterns were studied when Southern blots, carrying stallions DNA samples, were probed with eight synthetic tandem repeats (STRs), related by sequence variations in the basic unit. Because STRs preferentially crosshybridize with genomic VNTRs, they usually give patterns looking more like DNA fingerprints, but we found that even small modifications in the STR monomer could cause major changes in the hybridization profiles and could induce a shift of fingerprint pattern towards the detection of only one or two loci. This enables the use of STRs as direct genetic markers for linkage analysis, without cloning of the corresponding DNA fragment. Moreover, the set of STR variants can suggest consensus sequences allowing some prediction of the banding pattern.


A mouse genomic library in the bacteriophage P1 cloning system: organization and characterization.

Using the bacteriophage P1 cloning system, we have constructed a two to three times coverage, high-molecular-weight (HMW) genomic library from mouse C127 fibroblast cells. The library consists of about 127,500 clones with an average insert size of about 70 kb that are organized into 300 primary pools containing approximately 425 clones per pool. For screening purposes the primary pools are combined into secondary pools (approximately 4250 clones each) and tertiary pools (approximately 21,250 clones each). Screening is performed by the polymerase chain reaction (PCR) with DNA isolated from the secondary and tertiary pools. We have screened the library for 13 different mouse sequences and have detected 11. Clones generated from two of the eleven positive screens were isolated from the library (those containing the c-fos and G alpha i2 genes) and were further characterized. Direct double-stranded sequencing of DNA from P1 clones with primers bordering the insert provided sequence information from each end of the cloned DNA.


RASA contains a polymorphic microsatellite and maps to bovine syntenic group U22 on chromosome 7q2.4-qter.

The bovine gene for the p21ras protein activator (RASA) includes in its 5' untranslated region a (TG)n repeat. Analysis of this (TG)n repeat by PCR amplification of genomic DNA revealed a four-allele polymorphism. A cDNA probe was used to assign RASA to the region 2.4-qter of bovine Chromosome (Chr) 7 by in situ hybridization. PCR analysis of a panel of somatic hybrid lines allowed the assignment of RASA to the unassigned syntenic group 22 (U22) and thus localizes U22 on Chr 7.


Polymorphic microsatellite loci of the rat (Rattus norvegicus).

The EMBL and GenBank DNA databases were searched for microsatellite sequences of the rat containing dinucleotide repeats of (CA)n and (GA)n. Among those obtained, 23 sequences were analyzed by polymerase chain reaction to examine the size variation of the amplified fragment in inbred rat strains. All of the 23 microsatellite sequences varied in size among the strains tested. The 23 microsatellite loci in a pair of substrains separated from the same progenitor strain were then analyzed. Fragments identical in size were observed in all loci of the two substrains, indicating the stability of the microsatellite over a large number of generations. The microsatellite loci, therefore, should be useful markers for linkage analyses in the rat.


Deletion mapping of the mouse ornithine decarboxylase-related locus Odc-rs8 within Igh-V.

The Odc-rs8 locus belongs to a family of mouse DNA sequences related to the gene encoding ornithine decarboxylase (ODC). Odc-rs8 was mapped by recombinant inbred (RI) strain analysis to the region of Chromosome (Chr) 12 occupied by the variable region genes of the immunoglobulin heavy chain (Igh) complex. In the present study, alleles at Odc-rs8 were shown to cosegregate with those for Igh variable region (Igh-V or VH) genes among 37 inbred mouse strains that had been characterized previously for their haplotypes at Igh. For a more precise definition of the location of Odc-rs8 relative to Igh-V, DNAs from 17 Abelson murine leukemia virus (A-MuLV)-transformed pre-B cell lines cultured from mice heterozygous at Igh and Odc-rs8 were analyzed for the presence of DNA restriction fragments (RFs) derived from each parental Odc-rs8 allele. These cell lines, each of which has rearranged one or both Igh genes, previously were employed in mapping members of nine VH gene families by deletion analysis (Brodeur et al. 1988). Comparing the deletion profiles of the cell lines for Odc-rs8 with those for the VH gene families has located Odc-rs8b within the VHJ558/VH3609 gene cluster and Odc-rs8c either within or upstream of the 5'-most 9% of VHJ558, identifying Odc-rs8 as a potentially useful marker for the 5' end of the Igh complex.


Syntenic mapping and chromosomal localization of bovine alpha and beta interferon genes.

The previous assignment of bovine alpha-(IFNA) and beta-(IFNB) interferon gene families to syntenic group U18 was confirmed with additional cDNA probes and a bovine-rodent hybrid somatic cell panel representing all 29 bovine autosomal syntenic groups. Fluorescent in situ hybridization (FISH) localized these genes to bovine Chromosome (Chr) 8 band 15 and demonstrates that with biotinylated plasmids, as few as five tandemly arrayed sequences can be detected by conventional fluorescent microscopy. This technique can be applied to physical mapping of other multicopy genes in domestic animals.


Functional analysis of a t complex responder locus transgene in mice.

Transmission ratio distortion (TRD) of mouse t haplotypes occurs through the interaction of multiple distorter loci with the t complex responder (Tcr) locus. Males heterozygous for a t haplotype will transmit the t-bearing chromosome to nearly all of their offspring. This process is mediated by the production of functionally inequivalent gametes: wild-type meiotic partners of t spermatozoa are rendered functionally inactive. The Tcr locus, which is required for TRD to occur, is thought to somehow protect its host spermatid from the sperm-inactivating effects of linked distorter genes (Lyon 1984). In previous work, Tcr was mapped to a small genetic interval in t haplotypes, and a candidate gene from this region was isolated (Tcp-10bt). In this work, we further localize Tcr to a 40-kb region that contains the 21-kb Tcp-10bt gene. A cloned genomic copy of Tcp-10bt was used to generate transgenic mice. The transgene was bred into a variety of genetic backgrounds to test for non-Mendelian segregation. Abberrant segregation was observed in some mice carrying either a complete t haplotype or a combination of certain partial t haplotypes. These observations, coupled with those of Snyder and colleagues (in this issue), provide genetic and functional evidence that the Tcp-10bt gene is Tcr. However, other genotypes that were predicted to produce distortion did not. The unexpected data from a variety of crosses in this work and those of our colleagues suggest that elements to the TRD system and the Tcr locus remain to be identified.


Distortion of transmission ratio by a candidate t complex responder locus transgene.

The mouse t complex responder locus (Tcr) is centrally involved in the phenomenon of male-specific transmission ratio distortion (TRD) through its action in haploid germ cells. Previously, we identified a candidate gene, Tcp-10b, whose t allele generates alternatively spliced transcripts. The full-length Tcp-10bt transcript is present in pre- and postmeiotic germ cells and encodes a product that is virtually identical with that encoded by the wild-type allele. The alternatively spliced t-specific transcript is observed in post-meiotic haploid spermatids and would encode an altered polypeptide that could convey the Tcrt phenotype. To assess their function, we have introduced constructs representing each Tcp-10bt transcript into transgenic mice. Breeding experiments demonstrate that these two constructs alter the transmission ratios of t haplotypes from male mice, but in opposite directions. The results provide support for the hypothesis that Tcp-10bt is a component of the Tcr locus.


Four-state models and regulation of contraction of smooth muscle. I. Physical considerations, stability, and solutions.

Cyclic four-state models are frequently used in biology to represent a variety of molecular behaviors. A common experimental strategy to test such models is to follow the behavior of the real system after some of the rate constants are changed in a stepwise manner. We analyze the mathematical behavior of a simple example of such a model applicable to the regulation of contraction of smooth muscle, but our results apply in general to any linear, cyclic four-state model. We discuss detailed balance and requirements for linearity. We find that the only way to have sustained oscillations is for the rate constants of the model to be themselves oscillatory. We state conditions for decaying oscillations and find that in models that do not follow strictly first-order kinetics and do not satisfy detailed balance, these conditions can hold. We show analytically that the response of any state to step changes in the rate constants is the sum of three weighted exponentials plus a constant term, the steady-state value. We provide explicit expressions for the time dependence of all state variables. We discuss a simple way to use these results to obtain numerical solutions in cases where the rate constants change in an arbitrary way.


An analysis of life history evolution in terms of the density-dependent Lefkovitch matrix model.

The evolution of demographic characteristics is considered in terms of the density-dependent Lefkovitch matrix model, which describes a species' population dynamics with a stage-specific pattern of reproduction and mortality. We obtain the invadability condition of a mutant-type into the wild-type population at the equilibrium state. The condition depends on the left and right eigenvectors at the equilibrium state. The condition depends on the left and right eigenvectors at the equilibrium state and the difference, between wild-type and mutant-type populations, of the values of elements in the Lefkovitch matrix at the equilibrium state. It is also shown that if elements of the density-dependent Lefkovitch matrix are decreasing functions of population density, then the equilibrium population density increases in the process of natural selection; that is, K-selection acts even on the stage-structured population. The evolution of life history in perennial plants is discussed through two models as an application of the above results. The evolution of perennial plants with no vegetative reproduction is analyzed in the first example. It is shown that whether monocarpic perennials (which reproduce once and die) or polycarpic perennial plants (which reproduce more than once) are favored depends on the cost of a produced seed. The second example concerns perennial plants that reproduce vegetatively. It is shown that whether monocarpic or polycarpic perennial plants are favored depends on the cost of a seed and that where vegetative reproduction is common, polycarpic perennials with no seed reproduction are favored.


Four-state models and regulation of contraction of smooth muscle. II. Properties of the solutions and identification.

We analyze the behavior and the identification problem of cyclic four-state models. We find that for any state, or a weighted combination of two states, there can be at most one maximum, or one minimum, and two inflection points. We obtain necessary conditions for overshoot and undershoot and give examples. We describe procedures to estimate all the rate constants and discuss certain experimental aspects of the identification problem. Finally, we give an example of identification by obtaining the 10 model parameters from experimental data on skinned fibers from smooth muscle. These results, in conjunction with the results of the previous paper, can help in testing four-state models of regulation of contraction of smooth muscle and of a variety of other physiological and biochemical phenomena.


Improving Tikhonov regularization with linearly constrained optimization: application to the inverse epicardial potential solution.

Two methods to improve on the accuracy of the Tikhonov regularization technique commonly used for the stable recovery of solutions to ill-posed problems are presented. These methods do not require a priori knowledge of the properties of the solution or of the error. Rather they exploit the observed properties of overregularized and underregularized Tikhonov solutions so as to impose linear constraints on the sought-after solution. The two methods were applied to the inverse problem of electrocardiography using a spherical heart-torso model and simulated inner-sphere (epicardial) and outer-sphere (body) potential distributions. It is shown that if the overregularized and underregularized Tikhonov solutions are chosen properly, the two methods yield epicardial solutions that are not only more accurate than the optimal Tikhonov solution but also provide other qualitative information, such as correct position of the extrema, not obtainable using ordinary Tikhonov regularization. A heuristic method to select the overregularized and underregularized solutions is discussed.


Structured modeling of fish physiology.

The use of models in simulation and state estimation has proved useful in diverse applications, especially in industrial process control. The project presented here looked into the modeling of fish physiology for applications in fish physiology research and aquaculture. The models deal with gastric evacuation, metabolism, kidneys, gills, the cardiovascular system, and feeding behavior and are based on data from the literature. Model responses are mostly in accord with real responses in principle, but, as with most models of complex biological systems, the numerical accuracy is low in several cases. However, these structured models enable researchers to test hypotheses by altering the submodels and parameters.


Branched-chain amino acids and respiration.

A series of investigations suggest a specific role for BCAA in the regulation of respiration. In vitro incubation studies have shown that BCAAs improve the recovery of muscle force after fatigue. Further investigations revealed that leucine plays a key role in this action and acts in a manner not dependent on its use as an energy substrate. In humans, solutions enriched with BCAA have decreased PCO2 and stimulated the ventilatory response to hypercapnia, thereby corresponding to an enhanced ventilatory sensitivity with the administration of BCAA. The mechanisms for these actions are unknown. The most viable hypothesis is based on the ability of BCAA to decrease the synthesis of serotonin due to altered transport of AAs, including tryptophan, to the brain. Clinical studies have suggested a potency of BCAA in the treatment of respiratory dysfunction of preterm infants, as well as of patients with sleep apnea related to various disease states. The clinical applications of BCAA-enriched mixtures in respiratory diseases are still experimental, and many controversies exist concerning the validity of BCAA in clinical practice. Most TPN regimens contain BCAA approximating the average intake of BCAA in the Western diet. The question therefore remains whether additional BCAA supplementation is useful to achieve the suggested metabolic and pharmacological effects. Meticulous future studies are needed to establish the therapeutic value of BCAA in the treatment of various respiratory functions.


Management of diabetic patients requiring nutritional support.

A retrospective analysis was performed on 70 patients with diabetes mellitus who required nutritional support over the 10-yr period 1979-1989. Information was available for 65 patients, of whom 55 had non-insulin-dependent diabetes mellitus (NIDDM). Enteral nutrition (EN, 750-2200 kcal/day) was given to 40 NIDDM patients (group A) and 6 insulin-dependent diabetic (IDDM) patients (group B), and parenteral nutrition (PN, 1600-2400 kcal/day) was given to 18 NIDDM patients (group C) and 4 IDDM patients (group D). Three NIDDM patients required both types of feeding. Preadmission diabetes treatment remained the same during feeding for 31% of the total group (38% of group A, 33% of group B, 23% of group C, and 0% of group D). The NIDDM patients in group C who received insulin during PN required a high daily dose of approximately 100 U. The IDDM patients on PN required an increase of 225% from their preadmission daily dose. The likelihood of a patient requiring a major change from preadmission diabetes therapy depended mainly on the severity of the underlying illness and on the type of feeding (greater with PN) but not on preadmission therapy, age of patient, or type of EN (cyclic vs. continuous). Hypoglycemic episodes were uncommon in all groups. There were no significant differences between the prefeeding and feeding blood glucose levels and HbA1c results.


Performance of normal-weight and underweight men with marginal changes in energy intake.

This study compares the continuous response of six underweight (UW) (body mass index [BMI] < 18 kg/m2) and six normal-weight (NW) (20 < BMI < 25) men of similar age to a modest but sustained level of underfeeding and overfeeding. Habitual energy intake over 4 wk, body composition, and basal metabolic rate (BMR) were measured under metabolic-ward conditions. NW subjects were heavier by 9 kg and had 5% more body fat than UW subjects. The average BMR of UW subjects was 7.5% lower than NW subjects in absolute terms and also per kilogram fat-free mass per day but was higher by 8% when expressed per kilogram body weight per day. Three NW and three UW subjects were given a diet with 10% less energy than their habitual intake for 4 wk. They were brought back to the normal level of feeding for another 4 wk. Finally, they were overfed by 10% for 4 wk. This sequence was reversed in the remaining six subjects. Changes in body weight, BMR, and energy balance were assessed. UW subjects showed a quick and vigorous reduction in BMR (13.4%) during the 1st wk of underfeeding compared with NW subjects (8.1%). In the later weeks, the reduction was 8% in UW and 7% in NW subjects. Furthermore, UW subjects showed a tendency to resist a decrease in body weight (mean loss 180 g), unlike NW subjects (mean loss 730 g). With overfeeding, the mean increase in BMR for UW was higher (7.4%) than for NW (5.3%) subjects.(ABSTRACT TRUNCATED AT 250 WORDS)


Dietary fat level as determinant of protein-sparing actions of structured triglycerides.

This study assessed the effects of total parenteral nutrition (TPN) containing long-chain triglycerides (LCTs), an equimolar physical mixture of LCT and medium-chain triglycerides (MCTs), and a structured triglyceride synthesized from equimolar amounts of MCT and LCT on energy and protein metabolism after thermal injury (25% body surface area full-thickness scald burn). Male Sprague-Dawley rats (245-271 g) received isovolemic diets intravenously that supplied 250 kcal.kg-1.day-1, 2 g amino acid nitrogen.kg-1.day-1, and 50% of nonprotein calories as lipid and 50% as dextrose for 3 days. Whole-body and tissue leucine kinetics were estimated by a 4-h continuous infusion of L-[1-14C]leucine on day 3. Nitrogen balance, plasma albumin, plasma glucose, energy expenditure, and whole-body and liver and rectus muscle protein kinetic parameters were determined. No significant differences were noted in any of the parameters measured. This study suggests that the unique protein-sparing actions usually associated with structured triglyceride administration are not seen when they are provided as 50% of nonprotein calories. In addition, the ratio of MCT to LCT in the starting mixture from which the structured triglycerides are synthesized may be an important determinant of the protein-sparing actions attributed to these lipids.


Factorial experiment to determine influence of fish protein and fish oil on serum and liver lipids in rabbits.

Rabbits were fed purified diets consisting of casein (CA), fish protein (FP), and soy protein (SP) combined with MaxEpa oil (ME) or corn oil (CN) to determine the effects of dietary protein and lipid sources on serum total, lipoprotein, and hepatic lipid levels. Dietary proteins and lipids exerted significant (p < 0.05) separate effects on serum total cholesterol (TC) (p < 0.005), very-low-density lipoprotein cholesterol (VLDL-C) (p < 0.001), and high-density lipoprotein cholesterol (HDL-C) (p < 0.001), whereas only dietary proteins significantly affected low-density lipoprotein cholesterol (LDL-C) (p < 0.001) and the LDL-C/HDL-C ratio (p < 0.05). Hence, FP induced serum TC (233 mg/dl), VLDL-C (22 mg/dl), and LDL-C (151 mg/dl) intermediary to hypercholesterolemic CA (TC, 319 mg/dl; VLDL-C, 57 mg/dl; LDL-C, 204 mg/dl) and cholesterol-lowering SP (TC, 129 mg/dl; VLDL-C 19 mg/dl; LDL-C, 84 mg/dl). The twofold rise in HDL-C on feeding FP (35 mg/dl), compared with CA (20 mg/dl) and SP (16 mg/dl), resulted in a drop in LDL-C/HDL-C to a level similar to that of SP groups. The cholesterol-lowering action of ME (188 mg/dl), in contrast to CN (266 mg/dl), was reflected mainly in VLDL (ME, 15 mg/dl; CN, 50 mg/dl) but also in HDL (ME, 16 mg/dl; CN, 31 mg/dl) fractions. Compared with CN, the significant (p < 0.05) ME-induced rise in serum and VLDL triglycerides was accompanied by a significant (p < 0.001) drop in lipoprotein lipase activity.(ABSTRACT TRUNCATED AT 250 WORDS)


Total parenteral nutrition: iatrogenic immunosuppression.

It may be necessary for future formulations of TPN to encompass functions other than mere provisions of nutrients. Although current solutions of TPN are well tolerated in most patients for short periods and remain an important adjunct to modern surgical therapy, these solutions have significant room for improvement. Designing the ideal artificial gut for the highly stressed patient may include reproduction of the neurological, endocrine, and immune responses that occur after the oral presentation of foodstuffs.


Differences in fracture pattern between an urban and a rural population: a comparative population-based study in southern Sweden.

Differences in the incidence of hip fractures have been reported between urban and rural areas. In this population-based study the characteristics of fracture patterns between the city of Malmö and the nearby rural district of Sjöbo were compared. A total of 782 individuals in Malmö and 486 in Sjöbo were invited to participate. Fracture history for all invited was registered. The odds ratio for fracture was higher in Malmö, particularly for women over 70. More than half of the urban women aged 70 had a history of a fracture. A fourfold increase in fracture prevalence between the ages of 60 and 70 was observed in women in Malmö, whereas the prevalence doubled in Sjöbo. The differences in fracture patterns between these two urban and rural communities may be explained by different lifestyles.


Comparison and investigation of bone mineral density in opposing femora by dual-energy X-ray absorptiometry.

A dual-energy X-ray absorptiometry (DXA) machine was used to measure the bone mineral density (BMD) of both femora in 760 female volunteers. Each volunteer completed a questionnaire and exclusion criteria were applied such that only 480 of these were considered normal subjects. The remaining 280 women failed to comply with the criteria and were considered 'abnormal'; their BMD results were analysed separately. Two abnormal subgroups, one with previous long bone fractures and one with radiologically diagnosed osteopenia, were studied. BMD values for femoral neck, Ward's triangle and trochanter were compared between the two femora in all the above groups. No dominance relationship was found when comparing left to right femur, averaged over any population studied, but large differences were found between the femora in individual volunteers. There was a high correlation between BMD in opposing femora of 0.91, 0.91 and 0.84 for the femoral neck, Ward's triangle and trochanter respectively. However, in normal subjects the percentage variation in these regions ranged up to 34%, 64% and 80% respectively at the different femoral sites. In addition, the normal population was divided into two subgroups, one in which the density difference between the femora was large, and the other in which the difference was statistically insignificant. The analytical and anatomical variations between these two groups were investigated. Only part of the difference appeared to be due to analytical problems and it seems that there is a genuine difference in femoral density. Poor correlation for femoral neck percentage density difference was found with average BMD, age, height and weight in the normal population.(ABSTRACT TRUNCATED AT 250 WORDS)


Epidemiology of hip fractures in The Netherlands: women compared with men.

In this retrospective study the epidemiology of hip fractures in The Netherlands was evaluated. The number of hip fractures over the period 1972-1987 was collected from the Dutch Centre for Health Care Information (Stichting Informatiecentrum voor de Gezondheidszorg, SIG). The demographic data were collected from the Dutch Centre for Statistics (Centraal bureau voor de Statistiek, CBS). The age-adjusted incidence of hip fractures for both women and men 50 years of age and over, increased linearly over the period 1972-1987. There was also a linear rise in the age-adjusted incidence of hip fractures in women and men 65 years of age and over, but the rise in incidence was equal in both sexes. After the age of 50 years women and men showed an equal proportional increase in the age-specific incidence of hip fractures by 5-year age groups. This proportional increase was independent of the calendar year studied during the period 1972-1987. The increase over time in the age-adjusted incidence of hip fractures in inhabitants 50 years of age and over is the result of changes that act well before the age of 50 years, because the proportional increase in the age-specific incidence of these fractures after the age of 50 years did not change over the period studied. The cohort analysis showed that the age-specific incidence of hip fractures in more recently born birth cohorts is higher than that in birth cohorts born long ago. This observation agrees with the other observations.(ABSTRACT TRUNCATED AT 250 WORDS)


Hip fractures in the elderly: a world-wide projection.

Hip fractures are recognized to be a major public health problem in many Western nations, most notably those in North America, Europe and Oceania. Incidence rates for hip fracture in other parts of the world are generally lower than those reported for these predominantly Caucasian populations, and this has led to the belief that osteoporosis represents less of a problem to the nations of Asia, South American and Africa. Demographic changes in the next 60 years, however, will lead to huge increases in the elderly populations of those countries. We have applied available incidence rates for hip fracture from various parts of the world to projected populations in 1990, 2025 and 2050 in order to estimate the numbers of hip fractures which might occur in each of the major continental regions. The projections indicate that the number of hip fractures occurring in the world each year will rise from 1.66 million in 1990 to 6.26 million by 2050. While Europe and North America account for about half of all hip fractures among elderly people today, this proportion will fall to around one quarter in 2050, by which time steep increases will be observed throughout Asia and Latin America. The results suggest that osteoporosis will truly become a global problem over the next half century, and that preventive strategies will be required in parts of the world where they are not currently felt to be necessary.


Fracture risk as determined by prospective and retrospective study designs.

Both retrospectively and prospectively designed studies consistently show low bone mass and/or bone mineral content (BMC) to be risk factor for low-trauma fractures in postmenopausal women. Along with the reports of such studies there has been concern expressed that BMC measurements overlap between fracture groups, i.e., some women with high BMC develop fractures and some women with low BMC do not. In these commonly used epidemiologic study designs, BMC does not discriminate between those who have and have not experienced the untoward event at some level of the exposure factor. The ability to discriminate is more properly determined by the sensitivity and specificity of the measured value. To contrast the concepts of risk and sensitivity, a nested case-control study was conducted within a 24-year cohort study of women at risk for osteoporosis. We found that for each 1.0 decrement of BMC z-scores, the adjusted relative risk for the prospective study design was 1.67, while the odds ratio obtained from the most recent BMC z-score measurements was 1.87. A receiver operating characteristic (ROC) curve, calculated from the nested case-control study data, showed that BMC z-scores, measured after low-trauma fracture, have both low sensitivity and low specificity to detect existing fracture status.


The apparent incidence of hip fracture in Europe: a study of national register sources.

The objective of this study was to examine the apparent incidence of hip fracture from discharge rates in European countries. A request was sent to the Ministries of Health in all European countries, asking for the number of hip fracture patients by age and sex, between the years 1983 and 1985. Seventeen countries responded. As expected, hip fracture was most frequently found amongst the elderly, particularly women. The incidence of hip fracture rose exponentially with age in both sexes. It was higher in women than men and there was a three-fold range between countries in the female to male sex ratio. There was an eleven-fold range in apparent incidence amongst women and a seven-fold range amongst men between the various countries. The highest incidence was found in the northern part of Europe and the lowest in the Mediterranean area. There was a significant positive correlation between the age-standardized incidence rates reported in men from each country and that in women. There was a larger difference in incidence between countries than between sexes, which suggests important genetic or environmental factors in the causation of hip fracture. The extent to which this reflects imperfect capture of data is uncertain but will be important to determine in order to identify reasons for differences and to enable confident projections of the future magnitude of this disorder.


The myth of coupling reagents.

Coupling reagents have been proposed for peptide synthesis ever since the introduction of the method because of the convenience of the procedure, which consists of peptide bond formation by addition of a specific condensing agent to the mixture of carboxyl and amine components. However, truly efficient and yet innocuous coupling reagents are hard to find. Most coupling reagents give rise to unwanted reactions, which can be traced to certain characteristic structural features in their molecules. Such features are illustrated with the intermediates generated by dicyclohexylcarbodiimide and N-ethyl-5-phenyl-isoxazolium-3'-sulfonate (Woodward's reagent K). Some side reactions can be avoided if peptide bond formation is carried out in two separate steps: activation of the carboxyl group of the carboxyl component in the form of an active ester, followed by the catalyzed aminolysis of the active ester.


Bioactive conformation of Arg-Gly-Asp by x-ray data analyses and molecular mechanics.

Analysis of well resolved x-ray crystal structure data of proteins (Brookhaven protein data bank) has been combined with molecular mechanics methods using MM2, to determine possible bioactive conformations for the sequence Arg-Gly-Asp, which is believed to be involved in interactions of adhesive proteins with the glycoprotein complexes on activated platelets. In spite of the very flexible nature of this tripeptide fragment, only three general classes of structures were obtained. In one of them, the guanidinium and carboxylate moieties are in hydrogen-bonding proximity, while in the other they are separated by distances of greater than 8 A. Recent literature on the binding affinity of some rigid platelet-aggregation inhibitors seems to support the latter class of structures to correspond to the bioactive conformation.


Synthesis and disulfide structure determination of agelenin: identification of the carboxy-terminus as an amide form.

Agelenin, a blocker of the presynaptic calcium channel, is a 35-amino acid peptide having six cysteinyl residues recently isolated from the venom of the spider Agelena opulenta. However, it has not been confirmed whether the carboxy-terminus of this peptide is a free acid or amide. To elucidate this, we synthesized both peptides by a solid phase procedure, and compared their elution profiles with that of the natural product on RP-HPLC. The retention time on HPLC as well as the biological activity of the synthetic peptide-amide was found to be identical with those of natural agelenin, confirming that the carboxy-terminus of agelenin is amidated. The disulfide structure of agelenin was also determined to be linked between 3-19, 10-24 and 18-34, by comparing the tryptic peptide linked by two disulfide bonds with those synthesized by the selective formation of disulfide bonds.


Automated peptide-resin deprotection/cleavage by a robotic workstation.

A robotic workstation has been constructed that automates the deprotection of peptides with trifluoroacetic acid-labile side-chain protecting groups and cleavage from acid-labile resins. The workstation was constructed around a Zymark robot arm and is integrated with a peptide synthesis workstation. Peptide resin samples are deprotected and cleaved with a trifluoroacetic acid/scavenger cocktail that accommodates all common protecting groups used with Fmoc chemistry. Aqueous-ether continuous extraction is used to remove the scavengers and reaction by-products. The apparatus cleaves a 50-500-mg sample of peptide resin every 2 hours and provides the peptides as an aqueous solution in 10% HOAc. Crude peptides obtained with this apparatus are free from residual scavengers and range in yield from 50%-80%. This method is applicable to all but the most hydrophobic peptides and minimizes human contact with the noxious cleavage reagents.