[An autopsy case of Fahr disease (infantile form)].

A 13-year-old girl with Fahr disease (infantile form) was reported. Her parents were consanguineous. Her elder sister had mental retardation and spasticity of the lower limbs, and died at 23 years of age. The patient suffered from infantile spasms at 3 month. She was bed-ridden, nonverbal, microcephalic and blind. Cranial CT revealed massive calcifications in the basal ganglia, periventricular white matter, dentate nucleus and cerebellar white matter. EEG showed a suppression-burst pattern. At 13 years, she died of pneumonia and hyperammonemia. Microscopic examination of brain showed perivascular non-arteriosclerotic ferro-calcinosis. The periventricular granules are 1-4 mu or 12 mu in diameter. This pathological change was observed only in the central nervous system above midbrain. No calcifications were found in the pituitary and the vessels of pia mater. Also a reduced ornithine transcarbamylase activity was found in the liver, which was probably not related with cerebral calcifications. Infantile form of Fahr disease is rare and may be heterogeneous in etiology. However, clinical manifestations and pathological findings were similar to those in previous reports of Fahr disease in childhood. It is one of the disorders causing infantile spasms.


[A case of childhood epilepsy with occipital paroxysms (CEOP) presenting particular EEG findings].

We reported a case of an eight-year-old boy with CEOP. His parents and sibling were in good health. There was no family history of epileptic and neurological disease. He had his first attack of unconsciousness with fixation of eye movement for a few minutes at the age of 7 years. After six months, he experienced attacks of vomiting followed by loss of consciousness and elementary visual hallucinations consisting of red and blue colors. Sometimes he complained of contraction of visual field for 10 to 20 seconds, as if a curtain had fallen following the visual hallucination of a bright light spot. At the age of eight years, he was admitted to our hospital for evaluation and therapy. Investigations included neurological examination, IQ, CT findings were normal. Visual evoked potential revealed more reduced amplitude in the left side than in the right. The EEG findings during the waking state showed continuous bilateral 1-2 c/s spike and wave complex discharges in occipital and posterior temporal areas. These discharges were immediately suppressed by eyes-opening in an illuminated room, but not in a dark room. However, during the light sleep stage, diffuse irregular spike and wave discharges appeared frequently with left side dominance. From the clinico-electrophysiological findings we diagnosed him as CEOP. In order to investigate the changes of the occipital spike and wave discharges by photic stimulation, we administered intermittent photic stimulation (IPS) for 10 seconds at each frequency between 1-30 flashes/sec (f/s) in a dark room.(ABSTRACT TRUNCATED AT 250 WORDS)


[A case of acute encephalopathy with symmetrical low density areas in the thalami on CT; serial CT and MRI findings].

A 5-year-old girl was admitted to our hospital because of fever, vomiting, diarrhea, convulsion and disturbance of consciousness. She was diagnosed as having acute encephalopathy, which is characterized by the symmetrical low density area in the thalami on CT. Serial MRI findings revealed the bleeding with edema in the thalami, and multifocal lesions with prolonged T1 and T2 relaxation time in the acute phase of the illness. Multifocal lesions were no longer found two months later. The sequential changes on the images about the bleeding in the thalami were not confirmed by simultaneous CT scans. The findings of the bleeding in the thalamic lesion are consistent with those of an autopsy report previously described and indicate the vascular involvement in the thalami in acute encephalopathy, which is characterized by the symmetrical thalamic lesions with the characteristic finding of low density on CT scans.


[MRI findings of moyamoya disease in children].

Magnetic resonance imaging (MRI) findings were evaluated in two cases with moyamoya disease. A twelve-year-old girl and a five-year-old boy were admitted to our hospital with the complaints of transient ischemic attack. They were diagnosed as having moyamoya disease by cerebral angiogram. MRI clearly presented cerebral infarction by T2-weighted imaging and multiple small round or tortuous hypo_signal areas around the basal ganglia by proton density weighted imaging. These findings were consistent with moyamoya vessels shown by cerebral angiogram. Moyamoya vessels were visualized more clearly on proton density weighted imaging than on T1-weighted imaging. MRI is less invasive than cerebral angiogram and repeated safely. It might play an important role in a follow-up study of morphological changes on moyamoya vessels.


[Orbicularis oculi electromyogram in a girl with ocular myasthenia gravis].

A 12-month-old girl with ocular myasthenia gravis was reported. Electromyograms (EMG) of the orbicularis oculi muscle were studied during a period of 16 months. The degree of the waning phenomenon on EMG correlated well with that of her clinical symptoms. The study of post-tetanic exhaustion after repetitive electrical stimulation at 20 Hz for 4 seconds, was the most sensitive technique in the evaluation of the function of neuromuscular junction. EMG showed the waning phenomenon in the initial worsening after a high dose methylprednisolone therapy, and at the symptom-free stage without medication after the steroid therapy. In our patient, orbicularis oculi EMG was useful in the estimation of the severity of ocular myasthenia gravis and the effect of its treatment.


Carbohydrates in oral epithelia and secretions: variation with cellular differentiation.

The oral mucosa has been extensively studied in the past, and has thus become a model for demonstration of dynamic changes in cellular glycosylation and many other cell products such as keratins in relation to epithelial differentiation. The extensive knowledge of the glycosylation pattern in normal cells is instrumental in evaluation of the often occurring aberrant glycosylation of carcinoma cells. The emerging developments in antibodies to the glycosyltransferases and probes for their genes will enable us to gain insight into the regulatory mechanisms leading to the apparently well-controlled glycosylation process in epithelia. The first example of such a correlation has been studied in oral epithelia.


Carbohydrate changes in squamous cell carcinomas.

Cell surface carbohydrates serve as differentiation and development markers characteristic of different cell and tissue types. The expression of these carbohydrate antigens is often significantly altered in tumours, particularly in those arising from epithelial tissues. Analyses of cell surface carbohydrates in stratified epithelium have shown a remarkable variation in glycosylation pattern in relation to terminal differentiation. The carbohydrate expression is altered in squamous cell carcinomas and in premalignant lesions. There is evidence that the expression of certain carbohydrate structures in the deep invasive part of the tumours is correlated with tumour prognosis. The change in carbohydrate expression can at present be explained by the lack of synthesis of specific glycosyltransferases. New evidence suggests that the expression of certain carbohydrate structures may be importance for the formation of metastasies.


Changes in the glycosylation pattern of histo-blood group antigens in benign, premalignant and malignant laryngeal epithelial lesions.

The glycosylation of epithelial cell surface antigens is an indicator of cellular differentiation, and changes in the pattern of expression are seen in different premalignant and malignant epithelial lesions. The purpose of this investigation was to study alterations in the sequential build-up of carbohydrate structures during development of malignancy in laryngeal epithelial lesions. Sixteen routinely processed biopsies showing grave dysplasia/carcinoma in situ were examined. All patients were observed without treatment until malignancy could be affirmed. Five of the patients normalized their laryngeal mucosa spontaneously during observation, whereas 11 progressed invasive carcinoma. Biopsies from normal, hyperplastic and carcinomatous laryngeal epithelium were used as controls. Six monoclonal antibodies (MAbs) related to the ABO- and the TTn blood group systems were used for demonstration of carbohydrate antigens by an indirect immunofluorescent staining method. Benign lesions had a normal sequence of glycosylation. Dysplasias showed accumulation of shorter chains in superficial layers, usually with a patchy dispersion. It is possible by carbohydrate distribution to objectively establish the diagnosis of grave dysplasias/carcinoma in situ, and on the basis of carbohydrate distribution we can divide the lesions into subgroups which are of prognostic relevance.


Carbohydrate antigens in human lung carcinomas.

Lung carcinomas represent a heterogeneous group of tumours with large variations in the biochemical, clinical, morphological and pathological manifestations. Despite the neuroendocrine features of small cell lung cancer (SCLC), it is today generally accepted that all types of lung cancer emanate from a common endodermally derived multipotent stem cell of the bronchial epithelium. NCAM (neutral cell adhesion molecule) has been shown to be a sensitive marker of SCLC. The presence of NCAM, with the alpha (2,8) polysialic acid units characteristic of embryonal NCAM, in SCLC and in a portion of NSCLC, seems to correlate with the malignant behaviour and prognosis of the tumours, suggesting that NCAM may have a functional role in the clinicopathological manifestations of lung cancer. Fuc-GM1 with 2-hydroxy fatty acids as a characteristic component of the ceramide has been found to be a unique ganglioside of SCLC, detected in the tissues as well as in serum from SCLC patients using specific monoclonal antibodies. Accumulation of sialylated and fucosylated polylactosamine type 2 carbohydrate glycolipid and glycoprotein antigens was found in serum and tissues of lung tumours in accordance with what is described in gastrointestinal carcinomas, and these antigens may be regarded as general carcinoma antigens irrespective of organ. Sialylated and fucosylated type 1 antigens, and gangliosides with NeuAc alpha (2,6)Gal linkage were also accumulated in lung cancer. The human immune system has been found to recognize lung cancer carbohydrate antigens, which might be human lung cancer autoantigens.


Simple mucin-type carbohydrate antigens (Tn, sialosyl-Tn and T) in gastric mucosa, carcinomas and metastases.

Immunohistochemical study of the expression of simple mucin-type carbohydrate antigens (Tn, sialosyl-Tn, T and sialosyl-T) was performed using specific monoclonal antibodies in the mucosa adjacent to gastric carcinomas histologically appearing unaffected (n = 58), and in primary gastric carcinomas (n = 87) and their metastases (329 lymph nodes and two liver metastases). Normal-looking mucosas: Tn antigen expression was identified in all the cases; sialosyl-Tn in eight cases; T antigen was never expressed and sialosyl-T antigen was observed in four cases; the expression of these antigens was distinctly limited to the cytoplasm, mostly in the supranuclear (Golgi) area. All the mucosas with intestinal metaplasia showed sialosyl-Tn expression in the goblet cells. Gastric carcinomas: 80 cases (91.9%) stained for Tn antigen, 69 cases (19.3%) expressed sialosyl-Tn antigen, 18 cases (20.7%) expressed T antigen and 17 cases (19.5%) stained for sialosyl-T antigen. In contrast to normal mucosa, carcinoma cells expressed simple mucin-type antigens both at the cytoplasm and at the cell membrane. Most primary carcinomas were concurrently stained for Tn and sialosyl-Tn antigens alone (41.1%), or together with T antigen or sialosyl-T antigen (28.7%). We found a close relationship between the expression of simple mucin-type carbohydrate antigens in the primary tumours and their respective metastases. T antigen (and sialosyl-T antigen) expression was correlated with the wall invasiveness of the tumours. The 18 tumours expressing T antigen and 16 out of the 17 tumours expressing sialosyl-T antigen had nodal metastases and/or sialosyl-Tn expression and the aggressiveness of the tumours (wall penetration, lymph node metastasis and venous invasion). No significant differences were observed between positive and negative tumours for Tn, sialosyl-Tn, T and sialosyl-T antigens regarding the morphologic at type, growth pattern, ploidy or lymphoid infiltrate of the primary tumours.


Basic biochemistry of cell surface carbohydrates and aspects of the tissue distribution of histo-blood group ABH and related glycosphingolipids.

Cell surface carbohydrates may be protein- or lipid-linked. The structural polymorphism of the oligosaccharide chains is extensive due to variations in monosaccharide composition, carbohydrate sequence, branching, linkage position and linkage anomericity. Blood group ABH and related glycosphingolipids show a remarkable tissue-specific expression with possible implications in areas such as transfusion medicine, transplantation surgery and oncology. This communication gives a condensed description of the present knowledge of the tissue-specific distribution of histo-blood glycolipids in humans.


Carbohydrate changes in bladder carcinomas.

The cloning of glycosyltransferases is a major breakthrough in the study of glycosylation in normal an malignant tissue. In bladder tumours we need a molecular explanation for the changes observed in especially Gal alpha 1-3 and GalNac alpha 1-3 transferase activity. Studies of the events (transcription/translation) taking place in the different cell layers would also be of importance, especially if the cells could be sorted layer by layer. The urine is a source of malignant cells, and could be a target for diagnosis as well as monitoring. Studies of urinary tumour-associated glycoconjugates or tumour-associated rearrangements in glycosyltransferase genes in exfoliated cells would be of great interest.


Genetic regulation of the expression of ABH and Lewis antigens in tissues.

Sequential appearance of ABH antigens in different animal species shows a progression from tissues of endodermal to ectodermal and finally mesodermal origin, human erythrocytes being the last cells to acquire these antigens. In view of this, ABH antigens should be called tissue or histo-blood group antigens rather than blood group antigens. In addition to the glycosyltransferases encoded by the ABO genes, several alpha-2, alpha-3 and alpha-4-fucosyltransferases are needed to account for the known ABH histo-blood group antigens. The genetic polymorphism of the genes encoding each of these enzymes defines inter-individual differences. In addition, in the same individual various tissues express these antigens in a different way. For each adult epithelial tissue, antigenic expression is related to cell maturation from germinal layer to surface epithelium. Differential expression is also found at various embryonal stages of the same cells. Examples of these phenomena are presented in an effort to gain further insight into the genetic regulation of the expression of these complex oligosaccharide molecules.


Carbohydrates of the cell surface: molecular aspects of glycosyltransferases and their genes.

With the development of monospecific antibodies to transferase proteins as well as cDNA probes to cloned transferase genes, the study of glycosylation and its regulatory mechanisms holds great promise for the future. Here we provide a brief overview of the present knowledge of glycosyltransferases, their primary structure, cellular topography, regulation, and the basis of the genetic polymorphisms as revealed by blood groups.


Carbohydrates of human immunodeficiency virus.

Elucidation of the mechanism by which viral infection induces the appearance of carbohydrate neoantigens is highly important. Results from such studies could be expected to be significant for a general understanding of the regulation of glycosylation, and perhaps especially important for the understanding of glycosylation in cancer. For anti-viral therapy in AIDS, inhibitors of glycosylation enzymes are very promising as their mode of action may preclude evolvement of resistent HIV substrains, which seems to be a common problem with the reverse transcriptase inhibitors presently used. Successful therapy with glycosylation enzyme inhibitors will, however, require the development of more specific and less toxic compounds. If carbohydrate antigens can elicit a neutralizing immune response in vivo, the possibility exists that carbohydrate neoantigens can be utilized in the construction of a vaccine against AIDS.


Nucleotide sequence analysis of an HTLV-I isolate from a Chilean patient with HAM/TSP.

Isolates of HTLV-I have been characterized from a number of different regions of the world; however, there has not been a nucleotide sequence analysis of an HTLV-I isolate from a South American country. Reported here is an individual from Chile identified with the HTLV-I-associated neurological disease HAM/TSP. The sera and the nucleic acid sequence of the HTLV-I present in peripheral blood lymphocytes from this Chilean HAM/TSP patient over a two year period are characterized. During this time, the patient's condition grew progressively worse. While the serological profile of this patient was unremarkable in comparison with other HAM/TSP patients previously described, nucleic acid sequence analysis identified two nucleotide positions which contained nucleotides unique to this Chilean isolate. The nucleotide sequence analysis also indicates that the Chilean HTLV-I isolate is more closely related to Caribbean and Japanese isolates of HTLV-I than to the African and U.S. isolates described so far.


Establishment of T-lymphoid cell lines from Morroccan patients with tropical spastic paraparesis.

Two T-cell lines were established from peripheral blood mononuclear cells of two Moroccan patients with tropical spastic paraparesis and then named PR52 and PR144. The two cell lines showed a T lineage of activated CD4+ with high density of Tac+ (IL2 receptor). No expression of CD8 was observed. The virus particles were detected by reverse transcriptase activity and the viral antigens were also detected by immunofluorescence (IF) and Western blot. After six months of culture greater than 90% of the cells exhibited HTLVI antigen by IF. Lysate virus particles on Western blot analysis revealed p19,p24, and p53 gag protein similar to those detected in C91/PL virus particles from an adult T-cell leukemia (ATL) patient. gp46 and gp61 were also weakly detected. These two T-cell lines established will serve as substrate for further comparative studies on TSP and ATL isolates.


Anti-human immunodeficiency virus effects of zidovudine in combination with double-stranded RNA poly I poly C in T cells and monocytes-macrophages.

A comparison of the activity against human immunodeficiency virus 1 of zidovudine (AZT) and poly I poly C double-stranded RNA both alone and in combination in MT4 cells and primary monocyte/macrophage (M/M) cultures was made. The inhibition of the HIV-induced cytopathic effect or reverse transcriptase production by AZT in MT4 cells was not modified by the combination of the two agents. In contrast, AZT inhibition of reverse transcriptase production in the supernatant of M/M cultures was enhanced by the addition of poly I poly C. The inhibitory effect of the drug combination was more marked in M/M than in MT4 cells, indicating that the evaluation of compounds involving the induction of an antiviral state should be tested not only CD4+ T cells but also in monocyte-macrophages.


Loss of synergistic response to combinations containing AZT in AZT-resistant HIV-1.

The use of combination chemotherapy for the treatment of HIV-1 infection offers promise for overcoming the problems of toxicity and development of resistance. Primary HIV-1 isolates from three patients before and after treatment with azidothymidine (AZT) were examined for sensitivity to AZT and synergistic response to three different combinations of drugs: AZT+fluorothymidine (FLT), AZT+dideoxyinosine (ddI), and FLT+ddI. All three patients initially harbored AZT-sensitive virus, but posttherapy isolates were resistant to AZT. The pretreatment, AZT-sensitive virus from each patient showed synergistic inhibition by the combinations of AZT+FLT, AZT+ddI, and FLT+ddI. In contrast, the posttreatment, AZT-resistant virus showed only addition or antagonism by the combinations containing AZT, whereas the synergistic response to the combination of FLT+ddI was preserved. Our study argues for early intervention with combination chemotherapy, since after development of resistance, AZT no longer showed synergy but addition or antagonism in combination with other drugs. After resistance to AZT has developed, combination chemotherapy not involving AZT may offer advantages over continued mono- or combination therapy involving AZT.


Glutathione and N-acetylcysteine suppression of human immunodeficiency virus replication in human monocyte/macrophages in vitro.

Glutathione (GSH), its derivatives and N-acetylcysteine (NAC) inhibit the induction of HIV-1 expression in a chronically HIV-1-infected promonocytic cell line (U1/HIV) and peripheral blood mononuclear cells (PBMC). We have examined the effects of GSH and NAC on HIV-1 replication in human primary monocyte/macrophages cultured in vitro. Ficoll-gradient purified human monocytes were cultivated in vitro for 7-10 days and then infected with HIV-1 (Bal and Ada-M). Infection was blocked or substantially reduced by GSH or NAC (5-20 mM). Significant reduction (greater than or equal to 90%) in the amount of virus released, as determined by measuring supernatant reverse transcriptase activity and secreted p24 protein, was obtained when the cells were treated for 4 h with greater than or equal to 10 mM of GSH or NAC. The inhibitory effects of GSH and NAC were concentration dependent. This anti-HIV-1 effect persisted in these cultures for at least 35 days without evidence of significant increase in HIV-1 expression. Thus, a single pulse exposure of HIV-1-infected monocyte/macrophages with GSH or NAC led to a sustained, concentration-dependent decrease in HIV-1 p24 antigen levels, as well as, reverse transcriptase activity without producing detectable cellular toxicity in monocyte/macrophages.


Plasma HIV-1 viremia in HIV-1 infected individuals assessed by polymerase chain reaction.

We established a method to estimate the amounts of HIV-1 particles in plasma from patients with HIV-1 infection by using polymerase chain reaction (PCR) following reverse transcription (RT) of viral RNA (RNA-PCR) and assessed the potential usefulness of this approach to monitor the changes of viral load in patients with AIDS or AIDS-related complex (ARC) receiving 2',3'-dideoxyinosine (ddI). Plasma samples were obtained from 77 patients with HIV-1 infection (49 AIDS/ARC and 28 asymptomatic seropositives). Following ultracentrifugation of plasma, RNA was extracted from the pelleted virus and subjected to RT and PCR. The number of HIV-1 virus particles in each sample was determined using known amounts of HIV-1 DNA as reference control for PCR. The current plasma RNA-PCR technique quantitatively detected HIV-1 particles in plasma from 76 of 77 (98.7%) HIV-1-infected individuals examined. The numbers of HIV-1 particles in plasma from patients with AIDS or ARC were markedly higher than those in plasma from asymptomatic seropositive individuals (p less than 0.0001). Higher levels of plasma HIV-1 particle numbers were detected in individuals with lower CD4+ T cell counts. Patients (n = 10) who received oral ddI at doses greater than or equal to 6.4 mg/kg/day for 8 to 14 weeks had a profound decrease in plasma HIV-1 particle numbers (p = 0.0051). Patients (n = 7) receiving ddI for 45 to 71 weeks also had a decrease (p = 0.018). It should be noted, however, that more research is required to evaluate the usefulness of this technique in assessing the disease status and monitoring the activity of antiretroviral therapy.


Quantitation of human immunodeficiency virus (HIV) with respect to disease stage and zidovudine (AZT) therapy.

Quantitation of HIV in 115 seropositive individuals was undertaken to evaluate the potential for HIV transmission as a nosocomial infection through the use of medical devices that may come in contact with the peripheral blood of HIV-infected individuals. The virus burden in the peripheral blood was estimated from the level of: plasma HIV p24 antigenemia; plasma viremia; p24 antigen in peripheral blood mononuclear cell (PBMC) lysates as indicators of productive infection; and frequency of latently infected cells. Negligible HIV levels were observed in the plasma and PBMC lysates of the majority of samples except for late-stage patients with certain opportunistic infections and/or lack of zidovudine (AZT) therapy. Some individuals on AZT therapy and at late-stage of disease may show antigenemia without plasma viremia or alternatively, plasma viremia may be observed without plasma antigenemia. PBMC lysate data indicated that the frequency of productively infected cells was less than one in 20,000 PBMCs for the majority of samples irrespective of status on AZT therapy or disease stage. HIV was detected in greater than 95% of the cocultures and within 14 days for most of the samples, again regardless of the stage of disease or status on AZT therapy. The frequency of latently infected cells in this cohort ranged from 125 to 3125 per million PBMCs and was calculated to be as high as 2.5% of the helpter T-cell (CD4+ cell) population in the peripheral blood. The average latently infected cell frequency was 2-3-fold higher in early stage patients not on AZT than in late-stage patients on AZT therapy.


Detection of HIV-1 genome in leukocytes of human colostrum from anti-HIV-1 seropositive mothers.

In order to obtain more information about the presence of HIV-1 in mononuclear cells of colostrum, research was carried out on both the HIV-1 genome in the cellular fraction of colostrum and the viral antibody in cell-free colostrum of eight anti-HIV-1 seropositive asymptomatic mothers. In five cases cell fractions of the colostrum harbored HIV-1 genome by DNA-DNA and DNA-RNA in situ hybridization, whereas viral antibody were detected in all cell-free colostrum specimens. The data confirms the colostrum as a possible route of HIV-1 infection.


Hyperimmunoglobulinemia in HIV-1 infected individuals does not clearly correlate with plasma levels of IL-6.

In this study we evaluated interleukin-6 (IL-6) plasma levels in 80 human immunodeficiency virus type 1 (HIV-1) seropositive (+) individuals and 51 HIV-1 seronegative (-) blood donors. Plasma IL-6, detectable only in a subset of HIV-1(+) individuals (45 of 80) and normal blood donors (28 of 51), was significantly (p less than 0.01) increased in HIV-1(+) subjects 187 +/- 20.5 vs. 86.3 +/- 14 pg/ml). Among HIV-1-infected individuals, ARC/AIDS patients showed the highest IL-6 values (243.3 +/- 43.3 pg/ml). HIV-1(+) subjects showed, at all the different stages of the disease, a significant increase in total gammaglobulins, particularly IgG (2071 +/- 101 vs 1265 +/- 34 of HIV-1 seronegative controls). Although among HIV-1-infected individuals, the group with detectable plasma levels of IL-6 shows the highest levels of IgG (2243 +/- 146 vs. 1790 +/- 105, p less than 0.05), no positive correlations were observed between plasma levels of IL-6 and total gamma globulins (r = 0.2) or IgG (0.17). IL-6 production was also examined in the endotoxin-free supernatants of peripheral blood cultured monocytes and CD4+ T lymphocytes, in the presence or absence of specific stimuli. The amount of IL-6 released in monocyte and CD4+ T-lymphocyte culture supernatants was similar in 40 HIV-1(+) individuals and 35 HIV-1(-) controls. Our data show that plasma levels of IL-6 are significantly increased in HIV-1-infected individuals, in particular in ARC/AIDS patients. However, such an increase does not strictly correlate with the degree of hypergammaglobulinemia in the same HIV-1-infected individuals.


HIV-1 sequence variation between isolates from mother-infant transmission pairs.

To examine the sequence diversity of human immunodeficiency virus type 1 (HIV-1) between known transmission sets, sequences from the V3 and V4-V5 region of the envelope gene from four mother-infant pairs were analyzed. The mean interpatient sequence variation between isolates from linked mother-infant pairs was comparable to the sequence diversity found between isolates from other close contacts. The mean intrapatient variation was significantly less in the infants' isolates then the isolates from both their mothers and other characterized intrapatient sequence sets. In addition, a distinct and characteristic difference in the glycosylation pattern preceding the V3 loop was found between each linked transmission pair. These findings indicate that selection of specific genotypic variants, which may play a role in some direct transmission sets, and the duration of infection are important factors in the degree of diversity seen between the sequence sets.


Mapping of immunodominant epitopes of the HIV-1 and HIV-2 integrase proteins by recombinant proteins and synthetic peptides.

Different parts of the human immunodeficiency virus type 1 and type 2 (HIV-1 and HIV-2) integrase proteins were expressed as TrpE fusion proteins in Escherichia coli and used to screen human sera. In the immunoblot, all HIV/integrase-positive human sera tested reacted with the carboxy-terminal third of the integrase protein. Furthermore, they crossreacted with the same part of the heterologous protein. Half (50%) of the HIV-1/integrase-positive sera additionally detected antigenic epitopes in the amino-terminal third of the HIV-1 protein. Two of the recombinant proteins were used to generate polyclonal rabbit sera, which react with type-common epitopes of both integrase proteins. To map the B-cell epitopes of the HIV integrase proteins in more detail, overlapping decapeptides representing the entire integrase proteins of HIV-1 and HIV-2 were synthesized and used in a pin-based oligopeptide ELISA to scan human sera. This method can define three potential immunogenic epitopes of the HIV-1 integrase and one potential epitope of the HIV-2 integrase. The immunodominant epitopes of the HIV-1 integrase, one localized in the amino-terminal (IDKAQDEHEKYHSNWRAM), one in the central (QMAVFIHNFKRKGGIGGY), and one in the carboxy-terminal (AVVIQDNSDIKVVPRRK) part of the protein were synthesized as oligopeptides and used to test a larger panel of human sera in ELISA (156 HIV-1+ sera and 104 HIV-1- sera). The amino- and the carboxy-terminal epitopes were of equivalent reactivity, while the central part of the HIV-1 integrase seems to be less immunogenic. Nearly 90% of the HIV-1/integrase-positive human sera could be detected by a combination of these three peptides.


[Manometric changes induced by antireflux surgery (Nissen) and its relation to pH measurement and clinical findings. An analysis 6 months after the intervention].

We analyzed the preoperative ambulatory 24 hour ph-metric and manometric characteristics in a group of 20 patients treated surgically for gastroesophageal reflux (PGER) by Nissen fundoplication. At 6 months post-surgery they were reevaluated instrumentally (manometric and 24 hour ph-metry) and clinically. All ph-metric parameters of PGER (total reflux time, clearance, number of episodes, number of episodes greater than 5 minutes duration and duration of longest episode) were significantly improved (p less than 0.01-0.05) post-operatively (as globally as by position supine and upright) and this improvement was associated with resolution of symptoms in 19 (95%) patients. Of the manometric parameters evaluated (lower esophageal sphincter pressure--LESP--and length--LESL, peristaltic, triphasic, biphasic, absent and simultaneous waves and relaxation of LES) surgery only produced improvement in the lower esophageal sphincter pressure (LESP) and length (LESL) (p less than 0.001). We conclude that antireflux surgery (Nissen) by improving the pressure and length of the LES is capable of producing clinical and ph-metric remission in almost all (95%) of patients studied six months after.


[The efficacy of Plantago ovata as a regulator of intestinal transit. A double-blind study compared to placebo].

The effect of Plantago ovata on patients with chronic constipation (CC) with or without irritable bowel syndrome (IBS) has been assessed by a double blind study comprising 20 patients with CC of which 10 had associated IBS. A clinical questionnaire, weight of feces and intestinal transit time measured with radiopaque markers were done. Patients were then randomly distributed, 10 receiving PO and 10 placebo. Similar tests were done after treatment one month later. All patients receiving PO had good results against only one in the placebo group. Frequency of stools increased from 2.5 +/- 1 vs 8 +/- 2.2 stools per week, p less than 0.001 for paired data). A decrease in consistency of stools was also observed in the treated group. Fecal weight and colonic transit time were not significantly modified in placebo patients, while weight increase was observed in the treated ones (124 +/- 71 vs 194 +/- 65, gr/d p less than 0.001 for paired data) as well as a decrease in transit time (48 +/- 15 vs 34 +/- 18 hours p less than 0.05 for paired data). No adverse effects were observed and particularly no flatulence as often seen in patients on bran.


[The action of 5-fluorouracil in DMH-induced carcinogenesis in rats].

An experimental model to observe the action of 5-fluorouracil on rats with DMH-induced cancer of colon was designed. Ten Wistar rats were injected weekly with 25 subcutaneous doses (21 mg/kg) of DMH, and another ten, coinciding with the last 10 doses of the former, received intraperitoneally 10 mg/kg of the cytostatic agent. The animals were sacrificed 7 days after the last injection and samples of the colon processed by the "Swiss roll" technique. Differences in the weight of the animals and in the localization, size and macro and microscopic type of the tumours encountered, as well as the localization and grade of the dysplasias are studied.


[The value of ADA in peritoneal tuberculosis].

The aim of this study was to confirm that ascitic fluid determination of adenosine deaminase activity (ADA) is useful for the diagnosis of tuberculous peritonitis. 109 patients with ascites have been studied; 4 had tuberculous peritonitis and 105 nontuberculous ascites. The mean value of ascitic fluid AQDA was 0.587 +/- 0.2 uKat/l in tuberculous peritonitis and 0.11 +/- 0.1 uKat/l in nontuberculous ascites (p less than 0.001). An ADA value upper than 0.40 uKat/l has a sensitivity of 100% and a specificity of 99% for diagnosing tuberculous peritonitis. Ascitic fluid determination of ADA is simple, cheap and has a good diagnostic accuracy. In countries with high incidence of tuberculosis, measurement of ADA in ascitic fluid should be used as screening test for tuberculosis.


[The effects of verapamil in experimental acute pancreatitis in the rat].

Experimental studies have shown that verapamil inhibits pancreatic exocrine secretion. In order to determine whether verapamil has any effect on acute pancreatitis (AP), we undertook an experimental study in Wistar rats. We used 72 animals divided into two groups. In all animals of both experimental groups, AP was induced by ligation of the biliary duct at its entrance in the duodenum. Animals were given saline (NaCl 0.9%), or 0.30 mgrs/hour verapamil. Subgroups of 9 animals, were treated for 6, 12, 18 and 24 hours; 6 animals group were then sacrificed, for biochemical studies (serum amilase, lipase, and calcium; and trypsin and chemotrypsin in the homogenized pancreas); the other 3 animals were used for morphologic study of the pancreas. Verapamil treatment decreased significantly tissue activity of trypsin (p less than 0.001) and chemotrypsin (p less than 0.0001) and increased serum lipase (p less than 0.05), and calcium. There was no statistical difference in serum amylase. Morphological findings include oedema, acinar necrosis, hemorrhage and vasculitis in non treated animals. Only oedema was observed in animals treated with verapamil. These results suggest a beneficial effect of verapamil on experimental AP induced by ligation of the bile duct in the rat.


[Esophageal carcinoma after the endoscopic sclerotherapy of varices].

We report the case of a 52-years-old smoking male, diagnosed of liver cirrhosis, who developed a squamous cell carcinoma of the esophagus 36 months after undergoing endoscopic injection sclerotherapy for bleeding esophageal varices. Nine courses with 3% polidocanol were performed along 10 months. It was injected intra and paravariceal at a total dose of 117 ml. The relationship between endoscopic injection sclerotherapy and developing squamous cell carcinoma of the esophagus is discussed.


[Solitary neurofibroma of the esophagus].

A case of solitary neurofibroma located in the mid-portion of the esophagus in a 61-year-old woman is reported. She was operated with the diagnosis of benign tumor of the esophagus (leiomyoma). The microscopic examination of the tumor tissue found nervous cells. The immunohistochemistry analysis confirmed neurofibroma.


[Eosinophilic gastritis: its spontaneous resolution].

A 34-year-old man presented with acute epigastric pain. Laboratory studies disclosed a total eosinophil count 7.7 x 10(9)/l. The pathological diagnosis was eosinophilic gastritis. There was a spontaneous resolution, and no treatment was required. Investigations for allergic disorders gave negative results.


[Anal polypoid tumor formation as the form of presentation of rectal gastric heterotopia].

Heterotopic gastric epithelium occurs in all portions of the alimentary tract, but it is extremely rare in the rectum. The authors report the finding of a pedunculated anal polyp in a 9-year-old girl with a six-month history of rectal bleeding. Microscopically, the polyp consisted of epithelium of the fundic type. Both the age of the patient and the finding of heterotopic epithelium of the fundic type alone supports a congenital malformation and not an inflammatory condition.


[Isolated mesenteric tuberculosis as the first manifestation of AIDS].

We report a case of mesenteric tuberculosis. Two painful abdominal masses and fever were the first manifestation of disease. Diagnosis was obtained by aspirative punction of a mesenteric adenopathy guided by ultrasonography. AIDS risk factors were not recognized by anamnesis but serum HIV antibodies were detected. Thus, extrapulmonary tuberculosis and the finding of HIV-Ab confirmed the diagnosis of AIDS in our patient. It is necessary to assay for HIV-Ab in extrapulmonary tuberculosis, especially when mesenteric tuberculosis is present.


[An analysis of a general prospective series of 3270 upper digestive hemorrhages].

The results of a global (general series of 3,270 episodes of upper gastrointestinal haemorrhage (UGH) admitted to our unit between the 15th of April 1983 and the 15th of April 1988 have been analyzed. All the patients entered a prospective protocol with 29 variables. Diagnostic and therapeutic approaches had previously been defined. The incidence of UGH in this area was 160 bleeding episodes/100,000 inhabitants/year. Mean age was 57 +/- 16.8 years and male/female ratio was 2.66/1. The percentage of patients older than 65 years was 33.85%. A history of non-steroidal anti-inflammatory drugs (NSAID) intake within 48 hours before the bleeding episode was obtained in 27.63%. Continued alcohol ingestion was observed in 25.96% and 34.37% of patients gave a history of a previous episode of bleeding. UGH presented with haematemesis and melena in 56% of cases, and 44% only with melena. On admission the bleeding was haemodynamically severe in 12.96% and a 19.69% of the patients had severe associated diseases. Early endoscopy in cases with UGH due to peptic ulcer revealed active bleeding in 16.35% (2.87% in jet and 13.48% oozing) and recent clot/visible vessel in 31.7%. The major causes of bleeding were peptic ulcer (54.31%), esophageal and gastric varices (10.73%) and acute lesions of the gastric mucosa (ALGM) (6.72%). Etiology of the haemorrhage could not be established in 8% of cases. Bleeding was persistent in 20.75% and limited in 79.25% of patients. Emergency surgery was needed in 14.43% of cases. The global mortality of the series was 7.65%.(ABSTRACT TRUNCATED AT 250 WORDS)


[The thalamic changes in progressive supranuclear palsy].

This report is to investigate neuropathological changes of the thalamus observed in progressive supranuclear palsy. Six autopsy cases were examined. They were 5 males and a female, whose age was between 57 and 72 years old. They developed dementia in various degrees and three of the cases showed periodic stuporous state and akinetic mutism in the last stage. The paraffined thin-sections were stained with hematoxylin-eosin, luxol-fast-blue, Holzer, modified Bielschowsky, Gallyas and anti-tau stainings. The histological changes were evaluated in the aspects of neuronal loss, fibrillary gliosis and neurofibrillary tangles. In conclusion, there were three types in six cases. The first was seen in two cases and that there appeared many Alzheimer's neurofibrillary tangles (NFTs) in medial nuclei, lateral and ventral nuclei as well as zona incerta and reticular nucleus. The second was observed in two cases and that the loss of nerve cells was severe, especially in medial nuclei and the dorsal part of lateral and ventral nuclei and fibrillary gliosis was observed beyond the lesions of neuronal loss. The third was that the appearance of NFTs was moderate in number and neuronal loss was minimal. Moreover, neuronal loss and NFTs of zona incerta were common in these cases. Argentophilic fibrillary inclusions were observed in the neuropil and astrocytes in all cases. It is important to investigate the correlations between neuropathological changes and clinical manifestations, especially in the peculiar type of dementia observed in PSP. The thalamic changes of PSP may play an important role in causing stuporous state and akinetic mutism as well as subcortical dementia.


[Changes in dementia after shunt surgery for normal pressure hydrocephalus].

In order to investigate the pathogenesis of normal pressure hydrocephalus (NPH), dementia was analyzed with regard to the characteristic preoperative picture, the time when dementia improved, and what aspect of dementia was improved, retrospectively. Twenty seven patients of NPH were divided into two groups; young adult group: patients under 64 years old and geriatric group: patients above 65 years old. We found that the only difference found between these two groups was severity in dementia grading, higher in geriatric group. Postoperative course or complications did not differ. Dementia in NPH was not an essential impairment of intellectual ability, but was characterized by the subset symptoms of dementia, such as decrease in spontaneity and interest, slowness and paucity of thought and action. Improvement of memory followed postoperative recovery of these subset symptoms which was observed at earliest three days after shunt surgery. We postulate that recovery of dementia in NPH is associated with not only a decrease in intracranial pressure but also an improvement in the microcirculation of the brain or an improved metabolism.


[Correlation between the analgesic effect by thalamic relay nucleus stimulation and somatosensory evoked potentials recorded from thalamus].

Electric stimulation of the thalamic sensory relay nucleus (Vc) has an analgesic effect on deafferentation pain, however, the analgesic effect differs from patient to patient. Electrode position and state of the substrate stimulated are considered important factors influencing the analgesic effect. In order to determine the best position for the stimulating electrodes, we recorded somatosensory evoked potentials (SEPs) from stimulating electrodes implanted in the Vc and compared thalamic SEPs with the analgesic effect of Vc stimulation. The subjects were thirteen patients with deafferentation pain, four patients with thalamic lesions, seven patients with suprathalamic lesions and two patients with infrathalamic lesions. We inserted the electrode array into the Vc stereotactically, and fixed it so that stimulation-induced paresthesia would cover the painful frea. The electrode array consisted of the four contact points of four electrodes spaced at 2 mm intervals within 10 mm from the tip. Using bipolar combinations of the four electrodes (twelve combinations in all), we stimulated the Vc for about half an hour with each combination. We then rated the degree (%) of analgesia as 100% when pain disappeared and 0% when there was no change. Thalamic SEPs elicited after stimulation of the contralateral median nerve were recorded from all four contact points simultaneously. The latencies, amplitudes and recorded positions of large early positive components (P1) followed by large negative components (N1) with latencies between 10 and 20 msec were then analyzed and compared with the best electrode combination for optimal pain relief and with the degree of analgesia.(ABSTRACT TRUNCATED AT 250 WORDS)


[Neuritic degeneration in Alzheimer-type dementia, with special reference to the presence of dark neurites].

Four cases of neuritic degeneration in the cerebral cortex of Alzheimer-type dementia patients have been examined by routine and Gallyas-silver electron microscopy. This examination has revealed neuritic changes described below. (1) Dendrites and axons with an increased electron density (dark neurites) were found scattered throughout the neuropils, these dark neurites thought to be either degraded neurites resulting from simple neuronal atrophy or retrograde and/or transsynaptic degeneration due to an axonal injury within the senile plaques. (2) Both dystrophic neurites and neuropil threads were found scattered throughout the neuropils and showed some similarities in producing abnormal substances in their processes and in their manner of aggregating, especially within the senile plaque region. These degenerated neurites may have been caused by neurotoxicity of the amyloid. On Gallyas electron microscopic inspection, neurites often are found to have small fibrous and sometimes tubular structures on which silver particles are deposited. This finding may represent the initial stage of a neurofibrillary change. (3) The regeneration of neurites though such ultrastructures is not known, but such concept is believed by many authors and is discussed.


[A patient with gigantic heterotopic gray matter with epileptic seizures].

We report a patient with partial seizure and gigantic heterotopic gray matter. A 23-year-old young man was admitted to our hospital with complaints of frequent epileptic seizures and psychiatric symptoms. There was psychomotor delay in infancy. At the age of 4 years, afebrile convulsions appeared on several occasions. Seizures characterized by a lapse of consciousness started at the age of 13 years. He often stayed in a fantasy world and became very emotional at such time. Cranial CT demonstrated an irregularity in the wall of right lateral ventricle and disappearance of the posterior horn on the same side. This lesion, adjacent to that wall, had a signal intensity that was similar to that of the gray matter on each sequence in MRI. Histopathology of this lesion showed a number of large and small neurons. Therefore, heterotopic gray matter was diagnosed. MRI demonstrated wide cortices suggesting polymicrogyria in the right parietal lobe. Complex partial seizures with eye deviation to the left were recognized. Interictal EEG showed frequent high voltage spikes in the right temporal, fronto-temporal and parieto-occipital areas independently. Therefore, epileptic foci were thought to exist in or around those lesions.


[Unilateral choreic movements in idiopathic hypoparathyroidism].

We report a patient with idiopathic hypoparathyroidism associated with unilateral calcification of the basal ganglia and contralateral choreic movements. A 66-year-old woman was admitted to our hospital because of involuntary movements of the left limbs. Two years before the present admission, she had operations for bilateral cataracts. Eight months before admission, she had an onset of severe carpal spasms and pain in both arms and legs. These symptoms improved after one month's rest in a hospital. About a month before admission, she noted progressive difficulty in using her left hand, followed by difficulty in walking due to the abnormal involuntary movement of the left lower limb. Two weeks before admission her carpal spasms appeared again, bilaterally. On admission, she was alert. The general physical examination was normal. Neurological examination revealed choreic movements in the left hand, arm, leg and neck, and grimacing of the left face. She also complained of the spasms and pain in her hands, dysarthria and shortness of the breath. Chvostek and Trousseau signs were positive. Laboratory examination revealed marked decrease in serum calcium level (2.57 mEq/ml) and increase in inorganic phosphorus level (6.40 mEq/ml). Serum level of parathyroid hormone was less than 10 pg/ml. Ellthworth-Howard test was positive, in that the infusion of parathyroid hormone (100 u) elicited a marked increment of the urinary excretions of phosphorus and cAMP. X-ray examination of the spine revealed ossification of the posterior longitudinal ligament in the cervical region. CT-scans of the brain revealed calcification in the region of right globus pallidus and putamen.(ABSTRACT TRUNCATED AT 250 WORDS)


Physiologic capacity of well-developed collaterals in patients with isolated left anterior descending artery disease.

To assess the physiologic significance of well-developed collaterals, 34 patients, with isolated left anterior descending artery disease (LAD) and without overt prior myocardial infarction, underwent cardiac catheterization and exercise thallium-201 emission computed tomography. The patients were divided into 3 groups; 11 patients with 90% stenosis of the proximal LAD and without collaterals (group 1), 11 with 99% stenosis of the proximal LAD, and without collaterals (group 2) and 12 with a total occlusion of the proximal LAD which was completely filled by well-developed collaterals (group 3). On left ventriculography, shortening fractions of the anterior wall were significantly reduced in group 2 as compared to group 1 and 3 (group 1 vs group 2: p less than 0.01, group 2 vs group 3: p less than 0.05), which reflected the lower ejection fraction of group 2 (p less than 0.01 and p less than 0.05, respectively). The perfusion defects of the anterior wall on both the initial and the delayed images were severer in groups 2 and 3 than in group 1 (group 1 vs group 2 and group 1 vs group 3 on the initial image: p less than 0.01, for both, group 1 vs group 2 and group 1 vs group 3 on the delayed image: p less than 0.05, for both). However, recovery of the perfusion defects from the initial image to the delayed image was better in group 3 than in groups 1 and 2 (group 1 vs group 2 and group 1 vs group 3: p less than 0.05, for both). Therefore, coronary blood flow through well-developed collaterals was considered to be comparable to the flow through a diseased vessel with 90% stenosis at rest. During maximal exercise, blood flow through well-developed collaterals was considered to be comparable to the flow through a diseased vessel with 99% stenosis, although the blood flow through well-developed collaterals was considered to be better than that through 99% stenosis during the recovery period. These findings suggest that patients with well-developed collaterals must be treated like those with severe stenosis.


Pharmacokinetic analysis of antibody localization in human colon cancer: comparison with immunoscintigraphy.

The biodistribution and imaging characteristics of the 111In-labeled anti CEA monoclonal antibody ZCE-025 were studied in five patients with suspicion of colorectal carcinoma. Evaluation included antibody pharmacokinetics and assessment of antibody distribution in surgical specimen, making a comparison with whole-body imaging with a gamma camera. ZCE-025 localization in tumors was demonstrated by gamma-camera imaging in 4 of the 5 patients, corresponding to surgical findings. Persistent accumulation of 111In in the lymph nodes was observed in one patient, whereas surgical exploration of these lymph nodes showed no gross or microscopic evidence of metastases of colon carcinoma. Analysis of individual plasma by size exclusion HPLC showed two radioactivity peaks, labeled antibody and free DTPA. No transchelation of 111In to circulating transferrin was observed. The blood clearance was fitted to a two-compartment equation and its half-lives were found to be 10.8 +/- 8.7 h and 69.5 +/- 21.8 h for t1/2 alpha and t1/2 beta, respectively. Total urinary excretion averaged 0.3% of the injected dose/h with a small patient to patient variation. At 24 hrs postadministration the predominant radiolabeled species in urine was free DTPA. Thereafter, radioactivity in urine was partly present as a low molecular weight catabolic product. No apparent correlation between CEA content and uptake of 111In-ZCE-025 in tumors resected by surgery could be found. How 111In-labeled antibody is accumulated into tumors as well as into some nontumor tissues needs further study.


The causes and clinical significance of exercise-induced silent myocardial ischemia evaluated by ischemic range and intensity with exercise Tl-201 myocardial SPECT.

We investigated the causes and long-term prognosis of exercise-induced silent myocardial ischemia (SMI) by means of exercise Tl-201 myocardial SPECT (Ex-SPECT) in 97 patients with effort angina or old myocardial infarction (OMI). These patients were proven to have significant stenosis by coronary angiography. The subjects were divided into three groups based on the presence or absence of Tl-201 redistribution or angina during exercise testing. Group one consisted of 34 patients who had redistribution on Ex-SPECT and angina during exercise testing: the painful myocardial ischemia (PMI) group. The second group consisted of 38 patients who had redistribution on Ex-SPECT, but no angina during exercise testing: the SMI group. The third group consisted of 25 patients who had no redistribution: the RD (-) group. The ischemic range and intensity were quantified by the defect volume ratio (DVR) and defect severity index (DSI), respectively. Comparison of the DVR and DSI values for the PMI and SMI groups revealed that the DVR and DSI values for the SMI group were lower than those of the PMI group. Also the prognosis of the SMI group tended to be worse than that of the RD (-) group. Thus, we concluded that the SMI and PMI group should receive identical treatment.


Myocardial uptake of antimyosin antibody compared with serum myosin light chain I levels in patients with myocardial infarction.

Myocardial accumulation of In-111-antimyosin (InAM) was evaluated in comparison with circulating serum myosin light chain I (LCI) level at the time of InAM injection. Seventeen consecutive patients were studied at various stages ranging from 6 days to 34 days after myocardial infarction (MI). The infarct area was positive for InAM uptake in all patients (100%), and significant myocardial uptake was observed in 14 patients (82.4%). The intensity of InAM uptake correlated with the infarct location shown by ECG and CAG. In contrast, 12 patients (70.6%) had normal or undetectable serum myosin LCI levels, with 5 being normal (0.42-2.5 ng/ml) and 7 undetectable (0.42 ng/ml or less). Only 5 patients (29.4%) had elevated serum myosin LCI levels at the time of InAM injection, and this elevation was slight, ranging from 3.4 to 4.5 ng/ml (mean: 3.75 ng/ml). Among patients with undetectable, normal, and elevated serum myosin LCI levels, there was no significant correlation between InAM uptake and the serum myosin LCI level. Thus, even after the serum myosin LCI level has decreased to normal, InAM can still bind to cardiac myosin in patients with MI, presumably until there is complete recovery from the hibernating myocardium due to ischemic damage.


Preoperative evaluation of myocardial viability by thallium-201 imaging in patients with old myocardial infarction who underwent coronary revascularization.

The myocardial uptake and redistribution in thallium scintigraphy and the regional wall motion by echocardiography were evaluated by a semi-quantitative method in 42 patients who previously had myocardial infarction (50 target vessels) and underwent coronary revascularization. The aim of this study was to elucidate the significance of the initial image, delayed image and redistribution on thallium-201 scintigraphy for clinical diagnosis of the myocardial viability. As a semi-quantitative analysis, we used a bull's-eye display for thallium image and centerline method for echocardiographic wall motion, and compared the results before and after revascularization. As a result, the thallium grade improved postoperatively in all 17 areas which preoperative had showed redistribution, and also in 11 of the 32 areas without preoperative redistribution. The sensitivity, specificity and accuracy of preoperative thallium redistribution for predicting myocardial viability were 61%, 100% and 78%, respectively, when the postoperative improvement in the thallium grade was used as the standard. The postoperative probability of improvement in the thallium grade increased in proportion to the preoperative thallium grade (delayed image) (p less than 0.01). There was no correlation between the preoperative thallium delayed image and postoperative improvement in wall motion. Postoperative improvement in thallium image and wall motion could not be predicted from the preoperative wall motion. Thus, postoperative improvement in thallium images can be anticipated if redistribution is present on the preoperative thallium image, and the preoperative thallium delayed image is useful for predicting myocardial viability. Improvement in wall motion could not be predicted preoperatively by these methods.


Application of 67Ga for the estimation of reticulocyte production.

In order to estimate the production of reticulocytes, which have a larger number of transferrin receptors than erythrocytes, we used 67Ga which is exclusively bound to transferrin in the blood. The pattern of uptake of 67Ga by reticulocytes was quite similar to the time course of transglutaminase activity which might be involved in receptor-mediated endocytosis. The preinjection of Fe3+ decreased the uptake of 67Ga by reticulocytes. These results suggested that 67Ga in a transferrin-bound form was taken up by reticulocytes via receptor-mediated endocytosis. It was showed that the application of 67Ga is very easy and useful for the estimation of reticulocyte production.


Tc-99m-HMDP bone uptake quantification and plasma osteocalcin levels in hemodialysis patients--a preliminary study.

In this preliminary study plasma osteocalcin levels and Tc-99m-HMDP (Technetium 99m hydroxymetylene diphosphonate) bone uptake (BU) were measured in 10 chronic end-stage renal failure patients who were on maintenance hemodialysis. The aim of this study was to determine the correlation between bone uptake and osteocalcin-a sensitive and specific marker of osteoblastic activity. There was a statistically significant increase in both 20 and 180 minute uptake in the patient group (36 +/- 2.7 and 39 +/- 3.6) when compared to the normal volunteers (32 +/- 3.1 and 19 +/- 2.7). Plasma osteocalcin levels were also significantly high (24.5 +/- 5.6 ng/ml) when compared with normal values (6.5 +/- 2.3 ng/ml). The correlations between osteocalcin and 20 and 180 min BU were high (r = 0.62 and 0.72 respectively). In conclusion, our preliminary study suggests that, in hemodialysis patients, Tc-99m-HMDP bone uptake quantification is a sensitive and non-invasive method for showing increased osteoblastic activity.


Recognition and killing of tumour cells expressing heat shock protein 65 kD with immunotoxins containing saporin.

The expression of heat shock proteins (HSP) of the 65 kD family (groEL) has been observed by flow cytometry using murine monoclonal antibody (MoAb) anti-HSP 65 kD (ML30) on the surface of B (Daudi) or T (H9) lymphoma cells, on a monocyte cell line (U937) and also on a primary culture of a human pancreatic carcinoma (HPC). Moreover, the MoAb ML30 was coupled to Saporin 6, a ribosome-inactivating protein recovered from the seeds of Saponaria officinalis, to kill HSP-expressing cells with a specific immunotoxin. An indirect method using first MoAb ML30 and then anti-mouse IgG1 immunotoxin was also performed. With this method a human serum positive for HSP65-antibodies was tested using anti-human IgG1 or IgM immunotoxins. All cell lines were inhibited when preincubated with the specific immunotoxin directed to HSP65 (ML30 SO6), although H9 cells were susceptible to immunotoxin only after thermal stress. Daudi and HPC cells were inhibited both after long-term culture and when freshly explanted from SCID mice. Proliferation of the U937 monocytic cell line, that constitutively expresses high levels of HSP65 on the surface (as determined by flow cytometry), was completely inhibited (100% inhibition) by the ML30 SO6. However, not all tumour cells constitutively express high levels of surface HSP65, as determined by cytometric analysis. For this reason it was not always possible to obtain complete inhibition of cellular proliferation.


Chlorambucil-monoglutathionyl conjugate is sequestered by human alpha class glutathione S-transferases.

The spontaneous reaction of 110 microM chlorambucil (4-[p-[bis(2-chloroethyl)amino]phenyl]-butanoic acid; CHB) with 5 mM GSH at 37 degrees C in physiological phosphate-buffered saline for 35 min gave primarily the monoglutathionyl derivative, 4-[p-[N-2-chloroethyl,N-2-S-glutathionylethyl]amino]phenyl]-butano ic acid; CHBSG) and the diglutathionyl derivative, 4-[p-[bis(2-S-glutathionylethyl]amino]phenyl]-butanoic acid (CHBSG2) with small amounts of the hydroxy-derivatives: 4-[p-[N-2-chloroethyl,N-2-hydroxy-ethyl]amino] phenyl-butanoic acid (CHBOH) and 4-[p-[N-2-S-glutathionylethyl-2-hydroxyethyl]amino]phenyl]-butanoi c acid (CHBSGOH). The inclusion of approximately physiological amounts of human glutathione S-transferases (GSTs) A1-1, A2-2, P1-1, M1a-1a M3-3 or P1-1 (for nomenclature see Mannervik et al., 1992, Biochem. J., 282, 305) had little or no catalytic effect on these reactions as determined by loss of CHB. However, GTSs A1-1 and A2-2 were associated with a significant increase of CHBSG at the expense of CHBSG2 + CHBSGOH suggesting that these GTs sequestered CHBSG at the active site. This interpretation was supported by inhibition studies which showed that CHBSG was a pure competitive inhibitor of the activity of GSTs A1-1 and A2-2 towards 1-chloro-2,4-dinitrobenzene with Ki's of 1.3 and 1.2 microM respectively. GSH transferases P1-1 and M1a-1a were inhibited by CHBSG above 10 microM. Incubation of 2 microM CHB, a concentration which may be of more significance for chemotherapy, in the presence or absence of GST A1-2 (20-50 microM) showed catalysis of GSH monoconjugation equivalent to 18% of the spontaneous rate. However, the dominant effect again was the sequestration of CHBSG which reached 74.3 +/- 1.5 (SEM)% of the total reactants at 60 min compared to 28.9 +/- 0.3(SEM)% in controls. CHBSG, although possessing a potential electrophilic centre, showed no detectable alkylation of plasmid DNA but indirect evidence was obtained that it alkylated other cellular macromolecules. It is concluded that the contribution of GSTs to catalysis of CHB detoxication will depend on factors not previously considered, namely the relative molarities of CHB, CHBSG and GSTs, and the cellular capacity to excrete CHBSG to relieve product inhibition.


Photodynamic therapy of the normal rat stomach: a comparative study between di-sulphonated aluminium phthalocyanine and 5-aminolaevulinic acid.

Dysplasia in the upper gastrointestinal tract carries a risk of invasive malignant change. Surgical excision of the affected organ is the only treatment available. Photodynamic therapy has been shown to be promising in the treatment of early and superficial tumours and may be useful for the ablation of dysplastic mucosa. Because of the diffuse nature of the disease, such treatment would necessarily involve destruction of large areas of mucosa and it is desirable to confine its effect to the mucosa in order that safe healing can take place. By means of photometric fluorescence microscopy, we have studied the pattern of photosensitisation in the normal rat stomach using di-sulphonated aluminium phthalocyanine (AlS2Pc) and 5-aminolaevulinic acid (ALA) as photosensitisizers. AlS2Pc resulted in a panmural photosensitisation of the gastric wall with the highest level encountered in the submucosa. The mucosa and muscularis propria were sensitised to equal extent. Following light exposure, a full thickness damage resulted. ALA is a natural porphyrin precursor and exogenous administration gave rise to accumulation of protoporphyrin IX (PPIX) in the cells. The resultant pattern of photosensitisation was predominantly mucosal and its photodynamic effect was essentially confined to the mucosa. ALA produced a selective photosensitisation of the gastric mucosa for its photodynamic ablation with sparing the underlying tissue layers.


Interactions between human monocytes and tumour cells. Monocytes can either enhance or inhibit the growth and survival of K562 cells.

Human bloodstream monocytes can kill cultured tumour cells (K562), as assessed by specific release of 51Cr from the targets and by inhibition of 3H-thymidine incorporation. Confluent monolayers of monocytes were required for maximal cytotoxicity, and the density of the K562 cells was also an important factor. For example, when K562 cells were seeded at high cell densities, they were killed during incubation with monocytes, but when seeded at low cell densities their growth and survival was enhanced during culture with monocytes. The factor(s) which promoted the survival and division of low density K562 cultures was endogenously secreted from monocytes as it was present in monocyte-conditioned medium, whereas the cytotoxic factor(s) were only expressed during co-culture of monocytes with K562 cells. Conditioned medium from HL 60, U-937, HeLa and K562 could also enhance the growth and survival of low density K562 cultures, and a similar effect was also observed upon the addition of catalase and superoxide dismutase to such cultures. Thus, the monocyte:target ratio is important in determining whether monocytes exhibit cytotoxic or growth-promoting effects and hence tumour-derived or monocyte-derived reactive oxidant species may play a role in tumour cell cycle regulation.


A monoclonal antibody-beta-glucuronidase conjugate as activator of the prodrug epirubicin-glucuronide for specific treatment of cancer.

The anti-pan carcinoma monoclonal antibody (MAb) 323/A3, linked to E. coli-derived beta-glucuronidase (GUS) was used to study the tumour-site-selective activation of the prodrug Epirubicin-glucuronide (Epi-glu). Epi-glu was isolated from the urine of patients treated with Epirubicin (Epi) by reversed phase chromatography on a silica-C18 column. Epi-glu was stable in human blood and was not converted into Epi by A2780, MCF-7, or OVCAR-3 cancer cells, despite the presence of intracellular GUS. The stability of the prodrug was confirmed in BALB/c mice. MAb 323/A3 and GUS were linked through a stable thioether bond. The conjugate (1:1) was purified by ion exchange and gel filtration chromatography. Binding to target cells revealed an immunoreactivity of at least 60% and good retention of enzyme activity. A protein dye (sulforhodamine B) assay was used to analyse cytotoxicity. Epi (IC50 of 0.003-0.2 microM) was 100-1,000 times more toxic than Epi-glu (IC50 of greater than 20 microM), when cancer cells were exposed for 4 or 24 h to the drugs. The low cytotoxicity of Epi-glu was most likely due to the reduced cellular uptake rate of the prodrug (2.7 pmol 10(-6) cells min-1) as compared to that of the parent compound (25 pmol 10(-6) cells min-1). Pretreatment of antigen-positive cells with the 323/A3-GUS conjugate prior to prodrug exposure completely restored cytotoxicity as a result from hydrolysis of Epi-glu into Epi. Our results demonstrate that the 323/A3-GUS conjugate can specifically activate the stable non-toxic prodrug Epi-glu at the tumour cell level.


Radioimmunotherapy of human head and neck squamous cell carcinoma xenografts with 131I-labelled monoclonal antibody E48 IgG.

Monoclonal antibody (MAb) E48 reacts with a 22 kD antigen exclusively expressed in squamous and transitional epithelia and their neoplastic counterparts. Radiolabelled with 99mTc, MAb E48 is capable of targeting metastatic and recurrent disease in patients with head and neck cancer. In this study, the capacity of 131I-labelled MAb E48 to eradicate xenografts of human squamous cell carcinoma of the head and neck (HNSCC) in nude mice was examined. Experimental groups received a single i.v. bolus injection of 400 microCi MAb E48 IgG (number of mice (n = 6, number of tumours (t) = 9) or 800 microCi MAb E48 IgG (n) = 5,t = 7), whereas control groups received either diluent (n = 3,t = 5), unlabelled MAb E48 IgG (n = 4,t = 5) or 800 microCi 131I-labelled isotype-matched control MAb (n = 6,t = 9). A 4.1-fold increase in the median tumour volume doubling time and regression of two out of ten tumours (20%) was observed in mice treated with 400 microCi. In mice treated with 800 microCi. In mice treated with 800 microCi, two out of seven tumours (29%) showed complete remission without regrowth during follow-up (greater than 3 months). Median tumour volume doubling time in the remaining five tumours was increased 7.8-fold. No antitumour effects were observed in mice injected with diluent, unlabelled MAb E48 or 131I-labelled control MAb. In the same xenograft model, chemotherapy with doxorubicin, 5-fluorouracil, cisplatin, bleomycin, methotrexate or 2',2'-difluorodeoxycytidine yielded a less profound effect on tumour volume doubling time. Increases in tumour volume doubling time with these chemotherapeutic agents were 4, 2.2, 2.1, 1.7, 0, and 2.6 respectively. Moreover, no cures were observed with any of these chemotherapeutic agents. From the tissue distribution of 800 microCi MAb E48, the absorbed cumulative radiation doses of tumour and various organs were calculated using the trapezoid integration method for the area under the curve. To tumour xenografts, 12,170 cGy was delivered, blood received 2,984 cGy, whereas in every other tissue the accumulated dose was less than 6% of the dose delivered to tumour. These data, describing the first radiolabelled MAb with therapeutic efficacy against HNSCC, suggest radioimmunotherapy with MAb E48 to be a potential therapeutic modality for the treatment of head and neck cancer.


Classical oestrogen receptor is not detectable in pancreatic adenocarcinoma.

Recent reports have suggested the presence of oestrogen receptors (ER) in pancreatic carcinoma. Therefore the tumour may be sensitive to hormone manipulation therapy. We examined 23 biopsies of human pancreatic carcinoma tissue for the presence of ER. The tissue was assayed by two methods: Iso-electric focusing (IEF) and ER-ICA an immunocytochemical assay. All biopsies were tested with ER-ICA and ten by IEF. Each biopsy was assessed histologically for tumour content and 20 contained adenocarcinoma. None of the samples of pancreatic carcinoma were positive for ER using the ER-ICA, and none demonstrated the binding peak typical of ER in the IEF assay. These results suggest that in pancreatic carcinoma oestrogen receptor is either absent (or at very low levels), or of a different type to that found in breast and uterine tissue. On theoretical grounds at least, this must raise questions as to the oestrogen sensitivity of pancreatic cancer.


Interstitial laser hyperthermia: a new approach for treating liver metastases.

The palliative management of hepatic metastases remains unsatisfactory. There is a need for a simple non invasive technique which can stop or retard the rate of tumour growth. In principle, Interstitial Laser hyperthermia may fulfil such a role. In experimental studies, this technique produced precise in situ necrosis within solid organs which healed safely. In a pilot feasibility study, we treated ten patients with a total of 18 hepatic metastases on 31 occasions using a percutaneous approach to achieve an overall objective response rate of 44%. The treatment proved simple to perform, was well tolerated and produced radiological evidence of necrosis in small metastases (diameter less than or equal to 3 cm). However, further research is required before the technique can be regarded as established. Its future role in most cases will be to control the growth of discrete hepatic metastases unsuitable for resection. In instances where the extent of necrosis can be matched accurately to tumour volume, the potential for cure exists.


Prognostic significance of breast cancer axillary lymph node micrometastases assessed by two special techniques: reevaluation with longer follow-up.

Special techniques such as serial macroscopic sectioning (SMS) or immunohistochemical staining (IH) improve the detection rate of micrometastases but this detection is of value only if it improves the prediction of recurrence and survival. We first studied the prognosis of 120 patients with a single micrometastasis detected by SMS in a series of 1,680 primary operable breast carcinoma with a median follow-up of 7 years. A significant difference in recurrence (P = 0.005) and in survival (P = 0.0369) was found between node-negative patients and those with one single SMS micrometastasis, but SMS micrometastases were not a predicting factor by multivariate analyses according to the Cox model. We then studied the prognostic significance of patients with a micrometastasis detected by IH in node-negative carcinoma: 37 micrometastases from a series of 89 invasive lobular carcinoma (ILC) and 13 single micrometastases from a series of 129 invasive ductal carcinoma (IDC). In the ILC group, IH micrometastases had no prognostic value (median follow-up: 9.3 years). In the IDC group, IH micrometastases were correlated with recurrences (P = 0.01) and were the most significant predicting factor, but were less correlated with survival (median follow-up: 15.6 years). Three main points emerge from this study: (1) SMS micrometastases have a prognostic significance and macroscopic sectioning is recommended as a routine technique not requiring excessive work. (2) IH micrometastases in infiltrating lobular carcinoma have no prognostic significance. (3) The value of IH is debatable in infiltrating ductal carcinoma, since the technique is of principal use in predicting recurrences. It should therefore be carefully assessed vs other prognostic factors currently under study.


Tumour growth rate and DNA flow cytometry parameters as prognostic factors in metastatic melanoma.

The prognostic value of flow cytometric parameters and tumour growth rate of melanoma metastases under the mouse renal capsule was investigated for tumours from 117 consecutive patients referred to the Helsinki University Central Hospital Melanoma Team. DNA flow cytometry (FCM) was interpretable for the tumours of 114 patients, and growth rate analysis for 82 patients, both results being available from 79 patients. Thirty-six percent of the tumours were DNA diploid and 64% DNA aneuploid. Tumour ploidy and S-phase fraction were shown by multivariate Cox model analysis to be independent prognostic variables and major determinants of survival after first recurrence. Patients with DNA diploid or aneuploid tumours survived a median 16 and 27 months, respectively. A high growth rate of tumour sample in vivo under the mouse renal capsule tended to be a sign of poor prognosis, although not reaching statistical significance. Combining the results of FCM, tumour growth rate and TNM stage, we propose a highly efficient prognostic scoring method. Patients with a score above 0.75 had a median survival of 11 months compared to 30 months among patients scoring under 0.75 (P less than 0.0001). This score was the most significant (P less than 0.0001) prognostic factor in the Cox model when TNM stage, age, ploidy, SPF, and tumour growth rate were analysed as covariates.


CMV chemotherapy for advanced transitional cell carcinoma.

Between May 1986 and September 1990 a total of 43 patients with metastatic transitional cell carcinoma (TCC) of the urinary tract have been treated at our institution with combination chemotherapy (CMV) consisting of cisplatin 100 mg m-2 i.v. day 2; methotrexate 30 mg m-2 i.v. days 1.8; and vinblastine 4 mg m-2 i.v. days 1.8. Chemotherapy was recycled on day 22 and continued for a maximum of six cycles in responding patients. Of 33 patients with measurable disease 8 (24%) achieved a complete remission (CR). The median survival for patients achieving a CR was 13 months (range 5-29+) whilst the median survival for all 43 patients was 7 months (range 1-29+). Only three patients are still alive--two are disease free. More effective and/or less toxic chemotherapy regimens are needed for the treatment of patients with metastatic TCC.


An audit of the quality of cancer registration data.

The accuracy of cancer registration data in the East of Scotland (Tayside) Cancer Registry was audited by comparing 200 consecutive registrations (about 10% of the annual total) with the 'gold standard' of the Histopathology records. ICD codes were independently generated by a pathologist by examining final pathology reports and then compared to those codes given by the local cancer registrar. Discrepancies were graded by the pathologist and the epidemiologist according to severity. Major errors of coding were few. Minor and moderate differences in coding occurred because of the nature and structure of the coding system and the manner in which data are retrieved. The level of detail required by the Cancer Registry needs to be evaluated.


Expression of beta-2-microglobulin by nasopharyngeal carcinoma.

Serum beta-2-microglobulin (beta 2M) levels of 274 Chinese patients with different stages of nasopharyngeal carcinoma at presentation and that of 35 patients who developed distant metastases post-treatment were assayed. beta 2M level was found to increase with advancing stage of disease, with statistically significant differences among early-stage, advanced-stage, and metastatic disease. Elevated pre-treatment beta 2M levels were expressed more frequently by tumours with lower degree of histological differentiation. The sensitivity of serum beta 2M for diagnosis of nasopharyngeal carcinoma, however, is low.


Prognostic significance of p53 overexpression in gastric and colorectal carcinoma.

p53 expression was examined in 55 gastric and 107 colorectal carcinomas with an immunoperoxidase technique, using the polyclonal antibody CM1 on routinely fixed, paraffin embedded tissue. p53 protein was detected in 47% gastric and in 46% colorectal carcinomas and found to correlate with stage of disease and unfavourable clinical outcome (P less than 0.001). Thus, the proportion of positively reacting neoplasms increased as the stage progressed, tumours which had invaded regional lymph-nodes overexpressed p53 more frequently than localised carcinomas and an elevated level of p53 was associated with early relapse and death. In colorectal carcinoma p53 positivity was also linked with site and macroscopic configuration of the primary tumour and was most frequently expressed in carcinomas from the rectum and in ulcerative tumours. p53 overexpression was irrespective of tumour grade. Uniform negative reactivity with anti-p53 antibody was seen in normal epithelium adjacent to carcinoma, intestinal metaplasia, atrophic gastritis and in colonic adenomas. There was a good correlation between immunohistochemical staining on paraffin and frozen sections. These studies suggest that in gastric and colorectal carcinoma, immunohistochemical detection of p53 protein in routinely fixed tissue can be used along with other established parameters to assess prognostic outcome, especially to identify patients with poor short-term prognosis.


Trends in mortality rates from malignant melanoma in Sweden 1953-1987 and forecasts up to 2007.

To monitor mortality rates from malignant melanoma we analysed all patients in Sweden (6,324) who died of malignant melanoma in 1953 through 1987. Age-standardised rates per 10(5) increased from 1.1 to 4.0 in men and from 1.0 to 2.6 in women. The average annual increase levelled off in men from 4.6% during 1953-1967 to 2.0% in 1978-1987; and in women from 3.7% to 0%. Multivariate analyses showed that the change in rates for men was mainly due to a birth-cohort effect, whereas in women the rates changed similarly in all age-groups in accordance with a time-period effect. The risk of dying of malignant melanoma increased in men for birth cohorts up to 1932, whereas in women the rise continued for cohorts born as late as 1947. The best-fitted multivariate models were extrapolated to the year 2007, among men a slight increase in mortality rates seemed likely, whereas among women the rates will probably remain unchanged.


Occupation and bladder cancer: a death-certificate study.

Occupational statements on death certificates of 2,457 males aged 25-64 who died from bladder cancer in selected coastal and estaurine regions of England and Wales during 1965-1980 were studied. Excess mortality was found for deck and engine room crew of ships, railway workers, electrical and electronic workers, shoemakers and repairers, and tobacco workers. An excess of cases also occurred among food workers, particularly those employed in the bread and flour confectionary industry or involved in the extraction of animal and vegetable oils and fats. Use of a job-exposure matrix revealed elevated risk for occupations in which most workers were exposed to paints and pigments, benzene and cutting oils.


Tumour concentrations of flavone acetic acid (FAA) in human melanoma: comparison with mouse data.

Flavone acetic acid (FAA) showed impressive effects against murine solid tumours but no activity in clinical studies. The mechanism of action in mice may involve damage to tumour vasculature or immunomodulation, and these effects may be species-specific. Alternatively, concentrations of FAA achieved in mouse tumours may be higher than in human tumours. It is important to resolve this issue since it raises important questions about the relevance of in vitro versus in vivo tumour screens and the development of FAA analogues. As part of a Cancer Research Campaign Phase II study of metastatic melanoma in which 8.4 g m-2 FAA was given as a 6 h infusion, six tumour biopsies were obtained from four patients. FAA tumour concentrations were determined by HPLC and compared with subcutaneous murine solid tumours within the same analytical laboratory. Tumour/plasma percentages (range 26-61%; mean +/- SD, 43.9 +/- 11.4%) were similar to those in mice, as was the area under the curve (AUC) extrapolated to infinity and the AUC above the putative activity threshold of 100 micrograms ml-1. We conclude that the exposure of drug-refractory human melanoma tissue to FAA was comparable to that of sensitive mouse tumours. This suggests that reduced penetration of FAA into human tumours is unlikely to explain the lack of antitumour activity observed in clinical studies and that differences in mechanism of action are predominant.


Cancer mortality in the first degree relatives of young breast cancer patients.

In a retrospective cohort study, the mothers and sisters of 740 breast cancer patients aged under 36 at diagnosis have been studied for mortality and cancer incidence. Significantly increased breast cancer mortality was observed below age 60 (30 deaths; SMR = 3.4), but not at older ages (four deaths; SMR = 0.9). The cumulative breast cancer incidence in the relatives was 3.6% by age 50, 7.6% by age 60 and 11.6% by age 70. They also suffered excess mortality below age 60 for cancers of reproductive sites (cervix, ovary and endometrium; 15 deaths; SMR = 2.6) and lung (11 deaths; SMR = 3.2), but not for other sites (12 deaths; SMR = 0.9). This large population-based cohort study provides further confirmation of genetic susceptibility to breast cancer at young ages.


Localised lymphoma of bone: prognostic factors and treatment recommendations. The Princess Margaret Hospital Lymphoma Group.

Twenty seven adult patients with newly diagnosed non-Hodgkin's lymphoma localised to either bone (Stage IE) or bone and regional lymph nodes (Stage IIE) were treated between 1967 and 1988. Median age was 53 years and the commonest histology (21 patients) was diffuse histiocytic lymphoma. Twenty-four patients were treated radically: 15 with radiation therapy (XRT) alone and nine with chemotherapy plus radiation therapy (CMT). The cause specific survival for these patients was 56% at 5 years and 40% at 10 years. Survival was significantly better for patients treated by CMT (88% at 5 years) as compared to XRT alone (40% at 5 years, P = 0.03) and for age less than 60 (72% at 5 years) compared to greater than or equal to 60 (30% at 5 years, P = 0.018). Relapse-free rate was 27% at 5 years with XRT alone and 89% with CMT (P = 0.01). Risk factors for loco-regional relapse (seven cases) included: large tumour bulk, treatment by XRT alone and use of 'limited' radiation fields. No radiation dose-response relationship could be identified in this study. Long term local control and survival for localised lymphoma of bone were excellent after treatment by CMT but XRT alone was associated with unacceptably high local and distant failure rates.


Pseudotumor cerebri: risk factors, clinical course, prognostic criteria.

Eighty-five patients with the diagnosis of pseudotumor cerebri were retro- and prospectively studied. For clinical evaluation, the following criteria were used: neurological--headache, ophthalmological--visual acuity, visual field, ophthalmoscopy and psychic--memory. Each patient was assigned 2 categories of results expressed by: neuropsychic and ophthalmological scores which were statistically processed. The following conclusions related to the aim of our paper can be drawn: I. pregnancy (8% of cases), obesity (15%), generalized infections or located in the pelvic region+operated on (6%), hormonal treatment (cortisol (12%), estrogens (6%)); II. The clinical course was significantly favourable for: headache (51% of cases), defects of visual acuity (54%) and papilledema (63%). The long-lasting effects of the treatment applied showed: recovery (for the 2 parameters) in 27% of patients and improvement with sequelae for one and/or the other parameter in 73% of the cases; III. Among the more significant prognostic criteria were the ophthalmological symptoms and signs at onset of the disease. The ophthalmological sequelae (visual acuity, visual field defects, chronic papilledema and optic atrophy) are important factors in the prognosis of the disease and represent predictors for the surgical management.


Head injuries with temporal lobe compression.

The present study is based on the evolution of 219 cases of temporal compressive craniocerebral trauma admitted to the neurotraumatology department between 1985 and 1990. Correlations are made between the accident-operation time interval and the results obtained. Special emphasis is laid on the paraclinical methods used to make an accurate and complete diagnosis, of which computerized tomography is the most reliable for head injuries. The most frequent compressive lesions found in this series were the indirect cerebral lacerations. Their approach was done by a trephine opening aimed at the anterior 1/3 of the temporal lobe, in its basal part. The prognosis depends on the state of consciousness (patients in deep coma had small chances for survival) and on the time interval between accident and surgery, a rapid and adequately equipped transport of the patient being also of utmost importance.


Psychopathologic characteristics of the presuicidal syndrome.

The presuicidal syndrome (PS) is presented in this study with its prodromal symptoms which usually foresee suicide. One hundred patients with a PS were investigated taking into consideration all specific risk factors. Three stages of PS were outlined: psychic charging, marginal situation (Grenze-Situation) and psychic discharge. Interpretation of PS places it at the border between the clinical psychiatric fields and the ontological problems of humanity and leads to the understanding not only of the morbid psychic phenomenon in general and of the suicide in particular, but also to the major reasons of the human being who is trapped critical circumstances.


New insights into the regulation of megakaryocytopoiesis by haematopoietic and fibroblastic growth factors and transforming growth factor beta 1.

Effects of cytokines on murine megakaryocyte (MK) colony formation from either unfractionated marrow cells or purified early haematopoietic cells were studied. Recombinant interleukin-3 (IL3), interleukin-6 (IL6), granulocyte-macrophage colony-stimulating factor (GM-CSF), erythropoietin (Epo) and acidic and basic fibroblast growth factor (aFGF and bFGF) each was able to stimulate MK colony growth although they varied somewhat in their potential. IL6 and FGFs, in addition to their effect on MK colony growth, increased the size of individual MK. The combination of IL3 with IL6 or FGF resulted in an additive action. Monoclonal anti-IL6 antibody completely neutralized the activity of mouse IL6 and FGFs but had no effect on human IL6, mouse IL3 and GM-CSF. When using purified lineage negative marrow cells, only IL3 and IL6 promoted MK colony formation. Transforming growth factor beta 1 (TGF-beta 1) at 10-200 pg/ml selectively inhibited IL3-induced MK colony formation, and at 0.2-0.5 ng/ml it still had no obvious effect on the activity of IL6 or GM-CSF but caused an inhibition of FGF-induced MK colony formation. These data suggest that differential mechanisms are involved in the regulation of megakaryocytopoiesis by IL3, IL6, FGFs and GM-CSF, and that TGF-beta 1 negatively regulates MK development mainly by interfering with the action of IL3.


A PCR-based strategy to detect the common severe determinants of alpha thalassaemia.

A rapid and inexpensive polymerase chain reaction (PCR) based strategy is described which detects the three common, severe alpha thalassaemia determinants observed in southeast Asia (--SEA) and the Mediterranean (--MED and -(alpha)20.5). Oligonucleotide primers have been chosen which allow specific identification of both normal (alpha alpha) and abnormal (--) chromosomes using identical conditions in either the same or parallel PCR reactions. This strategy should be useful in the development of screening programmes to identify carriers of alpha thalassaemia (--/alpha alpha) and prenatal diagnosis of the Hb Bart's hydrops fetalis syndrome (--/--) for those populations in which this represents a major cause of perinatal death.


Lidocaine kindling does not generate epilepsy.

Forty Wistar rats were injected with a solution of lidocaine (90 mg/kg s.c.) 5 days per week for 30-40 days. In 36 of the animals, attacks of stiffness were obtained. After a period of disordered movements, the animals, remained completely immobile with the hindlimbs rigidly extended. The attacks of stiffness lasted from 10 to 60 minutes. The hemisection of the spinal cord at the thoracic level suppressed the rigidity of the ipsilateral leg. Electrical recording with electrodes applied to the cortex or implanted in the depth of the temporal lobe failed to reveal paroxysmal activity. These data could not confirm the results of other authors reporting typical epileptic seizures after lidocaine kindling. Despite essential differences, epileptic kindling and lidocaine-kindled stiffness attacks are both manifestations of the central nervous system plasticity. Therefore it appears that pharmacologically induced plasticity is a more general process than epileptic kindling.


Incidence of AIDS in HIV-1 infected thalassaemia patients. European and Mediterranean W.H.O. Working Group on Haemoglobinopathies and Cooleycare.

To estimate the cumulative incidence of acquired immunodeficiency syndrome (AIDS) in thalassaemia major patients (TMP) human immunodeficiency virus (HIV-1) infected through transfusion, 79 seropositive TMP were studied. At inclusion, mean age was 12.4 +/- 6.6 years; 40 were men; 21 were splenectomized. Centers for Disease Control, 1986 (CDC) stages and prescription of zidovudine were noted at least once a year. Cumulative incidence of AIDS and standard error were calculated using non parametric life table method. Age, sex, acute infection and splenectomy associations with progression to AIDS were tested using Breslow statistic. The median follow-up period was 4 years 11 months. At the end of the study period, 43 TMP were in CDC stage II, 23 in CDC stage III and 13 in CDC stage IV, including seven AIDS cases, of whom three had died. Four subjects died of other causes. Only two patients were treated with AZT prior to the occurrence of AIDS. Rate of progression to AIDS was not associated with acute infection, splenectomy, age, or sex. A cumulative AIDS incidence rate of 1.4% (SE 1.3%) was observed at 3 years and of 9% (SE 4%) at 5 years.


Development of a monoclonal antibody capable of differentiating platelet PLA1/PLA1, PLA1/PLA2 and PLA2/PLA2 genotypes.

A monoclonal antibody, LK-4, has been developed which distinguishes platelet PLA1/PLA1, PLA1/PLA2 and PLA2/PLA2 genotypes on platelet glycoprotein GPIIIa of Triton-solubilized platelet extracts. An ELISA assay has been developed which traps GPIIIa with Concanavalin A, enriching the platelet extract for the PLA antigens. A second monoclonal antibody, DEK-10, which reacts equally with GPIIIa of PLA1/PLA1 and PLA2/PLA2 platelet extracts is employed as an internal standard to correct for individual differences in GPIIIa content, GPIIIa extracted by Triton X-100 and GPIIIa trapped with Concanavalin A. This ELISA assay clearly differentiated 11 different PLA1/PLA1 subjects from eight PLA2/PLA2 women with a history of neonatal alloimmune thrombocytopenia as well as six unrelated obligate heterozygotes and should be useful in evaluating the PLA genotype of pregnant women and their families.


Melodic intonation in the rehabilitation of Romanian aphasics with bucco-lingual apraxia.

The main objective of the present study was to assess the efficiency of the melodic intonation therapy (MIT) in the rehabilitation of Romanian aphasics. Eighty predominantly Broca aphasics used the melodic intonation therapy when no other therapy methods were very efficient. The speech therapist intonated the respective word, then together with the patient and finally let him continue alone. The control group counted 80 aphasics and it applied other therapy methods. Each patient, regardless of the group, was tested twice, before and after therapy. Since most of the patients displayed severe language disorders, and other therapy methods failed in rehabilitating them, MIT was considered an efficient method in the early stages of Broca aphasia with bucco-lingual apraxia.


Evaluation of erythropoiesis after bone marrow transplantation: quantitative reticulocyte counting.

Erythroid regeneration is an important and separate element in the engraftment process in allogeneic and autologous bone marrow transplantation (alloBMT, autoBMT). Qualitative visual reticulocyte counting has proved inadequate in the evaluation of erythropoiesis after BMT but automated flow cytometry now allows the reliable quantitation of reticulocytes even to very low levels. Reticulocyte counts and highly fluorescent reticulocyte (HFR) counts (very early reticulocytes) were estimated daily in recipients of 22 autoBMT and 14 alloBMT using a Sysmex R-1000 automated reticulocyte counter. Marrow ablation caused an immediate and rapid fall in both the reticulocyte count and the HFR. Measurable numbers of reticulocytes persisted throughout the hypoplastic period, but HFR fell to zero in the majority of both the autoBMT and alloBMT. HFR rose significantly after a median time of 14 d post-autoBMT, and 12 d post-alloBMT. Attainment of 15 x 10(9)/l reticulocytes and 0.5 x 10(9)/l HFR at day 21 post-transplant was associated with ultimate engraftment in 100% cases. Inadequate engraftment was seen in the majority of patients whose responses fell below these levels. Graft-versus-host disease was associated with a transient slight reduction in reticulocyte count. Neither episodes of infection nor blood transfusions had any significant impact on trends of reticulocytes or HFR. Automated flow cytometric reticulocyte counting has been shown to provide an accessible measure of erythroid activity which may be of predictive value in the management of patients following bone marrow transplantation.


Molecular analysis of clonality in plasma cell dyscrasias.

It has been suggested that multiple myeloma, generally considered a neoplastic disorder of mature plasma cells, may arise from a pluripotent haemopoietic stem cell. The possibility that circulating lymphocytes derive from the same neoplastic progenitor has been tested in a large number of studies in the past few years, as proof of the interest that this subject is raising among scientists, and also of its elusiveness. We studied a group of 29 patients with plasma cell dyscrasias in order to evaluate clonality of haemopoietic cell populations. The X-linked markers hypoxantine phosphoribosyltransferase (HPRT) and phosphoglycerate kinase (PGK) disclosed no monoclonal component in seven heterozygous women. Analysis of immunoglobulin gene rearrangement with four probes showed a germline configuration in samples from 25/29 patients. Only four bone marrow samples from subjects with aggressive disease had rearranged C mu sequence; one had rearrangement of JH and C mu.