Orotic acid, a promoter of liver carcinogenesis induces DNA damage in rat liver

Orotic acid, a precursor of pyrimidine nucleotide biosynthesisand a promoter for liver carcinogenesis, when fed at 1% levelin a diet for 5 weeks resulted in liver DNA damage. The damagecan be monitored as alkali-labile lesions using alkaline sucrosegradients as well as alkaline elution technique. Furthermore,the induced DNA damage persists for up to three weeks afterwithdrawal of the orotic acid diet. The fad that several skin-tumourpromoters also induce DNA damage raises the question whetherDNA damage is a component in tumour promotion.     

Colonic carcinomas masquerading as Crohn's colitis.

When colonic carcinomas present with acute abdomen, the operating surgeon and the pathologist face a plethora of diagnostic and therapeutic problems. In this retrospective study of 92 cases of carcinoma colon, 4 presented with acute intestinal obstruction of which three had a turbulent post operative period and died. The resected colonic segment showed on gross examination cobblestone appearance characteristic of Crohn's disease but microscopically was ischemic with the stricture site showing features of an infiltrating poorly differentiated adenocarcinoma. We have made an attempt to study the various pathologic features and analyse their significance with reference to prognosis.     

An additional radial wrist extensor and its clinical significance

The knowledge of anatomical variations in the antebrachial and dorsal regions of the arm and hand are useful in hand surgery. The extensor carpi radialis intermedius and extensor carpi radialis accessorius are two classic variants described for the radial wrist extensors, in the antebrachial region. We report an additional extensor carpi radialis muscle taking origin from the common extensor origin, between the extensor carpi radialis longus and extensor digitorum communis. The tendon of the variant muscle divides below the abductor pollicis longus and becomes attached to the base of the second and third metacarpal bone. Due to its considerable size and independent origin from the lateral epicondyle, we suggest the present variation should be named extensor carpi radialis tertius. The clinical significance of the present variation is discussed.     

Comparing retinal sensitivities on blue-on-yellow and green-on-yellow perimetry in glaucoma suspects

Purpose:To compare the retinal sensitivities between the blue-on-yellow perimetry (BYP)/short-wavelength automated perimetry (SWAP) and green-on-yellow perimetry (GYP) among patients with and without nuclear sclerosis among glaucoma suspects.Methods:After ophthalmic examination, patients were subjected to two perimetric tests: BYP and GYP. The visual field (VF) parameters were compared between the two perimeters (p < 0.05 was considered significant).Results:Fifty-five eyes of 39 patients with a mean age of 60.53 ± 9.70 years were included in the study. Twenty-one eyes had clear lens or pseudophakia. Twenty-six eyes had lower grades of nuclear sclerosis (NO2NC2, NO3NC3) and eight eyes had higher grades of cataract (NO4NC4, NO5NC5). The mean retinal sensitivity (RS) in BYP was 22.08 ± 5.02 (dB) and in GYP was 23.84 ± 5.50 (dB) (p = 0.08). The mean defect in BYP was -2.56 ± 4.40 (dB) and in GYP was -3.24 ± 5.05 (dB), pattern standard deviation (PSD) in BYP was 3.65 ± 1.91 (dB) and in GYP was 3.83 ± 1.99 (dB), and foveal threshold (FT) was 24.20 ± 4.32 (dB) in BYP and 28.10 ± 4.50 (dB) in GYP. The two perimeters showed good agreement by the Bland–Altman plot for all parameters. Fourteen eyes showed perimetric changes suggestive of glaucoma by BYP. In these, GYP had a sensitivity of 92.86% (95% CI of 66.13% to 99.82%) and specificity of 95.12% (95% CI of 83.47% to 99.40%).Conclusion:BYP and GYP show good agreement. They are comparable in clear media as well as in different grades of nuclear sclerosis. GYP showed good sensitivity and specificity compared to BYP.     

Some factor VIII inhibitor antibodies recognize a common epitope corresponding to C2 domain amino acids 2248 through 2312, which overlap a phospholipid-binding site

The finding that human factor VIII (fVIII) inhibitor antibodies with C2 domain epitopes interfere with the binding of fVIII to phosphatidylserine (PS) suggested that this is the mechanism by which they inactivate fVIII. We constructed a recombinant C2 domain polypeptide and demonstrated that it bound to all six human inhibitors with fVIII light chain specificity. Thus, some antibodies within the polyclonal anti-light chain population require only amino acids within C2 for binding. Recombinant C2 also partially or completely neutralized the inhibitor titer of these plasmas, demonstrating that anti-C2 antibodies inhibit fVIII activity. Immunoblotting of a series of C2 deletion polypeptides, expressed in Escherichia coli, with inhibitor plasmas showed that the epitopes for human inhibitors consist of a common core of amino acid residues 2248 through 2312 with differing extensions for individual inhibitors. The epitope of inhibitory monoclonal antibody (MoAb) ESH8 was localized to residues 2248 through 2285. Three human antibodies and anti-C2 MoAb NMC-VIII/5 bound to a synthetic peptide consisting of amino acids 2303 through 2332, a PS- binding site, but MoAb ESH8 did not. These antibodies also inhibited the binding of fVIII to synthetic phospholipid membranes of PS and phosphatidylcholine, confirming that the blocked epitopes contribute to membrane binding as well as binding to PS. In contrast, MoAb ESH8 did not inhibit binding. As the maximal function of activated fVIII in the intrinsic factor Xase complex requires its binding to a phospholipid membrane, we propose that fVIII inhibition by anti-C2 antibodies is related to the overlap of their epitopes with the PS-binding site. MoAb ESH8 did not inhibit fVIII binding to PS-containing membranes, suggesting the existence of a second mechanism of fVIII inhibition by anti-C2 antibodies.     

Chronic mitoinhibition during promotion of hepatocarcinogenesis

We have reported previously that orotic acid (OA), a precursor for pyrimidine nucleotide biosynthesis, is able to promote carcinogenic process in both liver and duodenum of rats. The present study investigates the possible role of mitoinhibitory effects of OA as being responsible for its promotional effects. Male Fischer 344 rats were given a semisynthetic basal diet (BD) or a diet containing 1% OA for four weeks coupled with 2/3 partial hepatectomy (PH), and all animals were then continued on BD for an additional four weeks. This protocol is known to exert a promoting effect on the initiated rat liver. Livers were perfused, and the labeling index (LI) of isolated cultured hepatocytes was monitored. Hepatocytes isolated from livers of rats fed a BD or 1% OA exhibitedin vitro an LI of 39±2 and 24±1%, respectively. The loweredin vitro LI was seen even upon exposure to epidermal growth factor (EGF) (67±2% in OA-treated livers compared to 91±2% in hepatocytes from control rat liver). A similar four-week exposure to OA coupled with PH decreased hepatic DNA synthesis induced by a choline-deficient dietin vivo by about 50%. These results indicate that OA is able to decrease the response of normal hepatocytes to growth factors and suggest a possible mechanism of chronic differential mitoinhibition as a basis for promotion induced by OA.This work was supported in part by USPHS Grant CA 37077 from the National Cancer Institute, USA, by Grants from the National Cancer Institute, Canada and from the Ministry of Labour and Occupational Health, Ontario, Canada. E.L. was supported by a fellowship from Ministry of Education, Italy.Presented at the Proceedings of the International Meeting on Normal and Neoplastic Growth in Hepatology, Bari, Italy, June 1989.     

The Singapore Heart Failure Outcomes and Phenotypes (SHOP) Study and Prospective Evaluation of Outcome in Patients With Heart Failure With Preserved Left Ventricular Ejection Fraction (PEOPLE) Study: Rationale and Design

BackgroundHeart failure (HF) with preserved ejection fraction (EF) accounts for a substantial proportion of cases of HF, and to date no treatments have clearly improved outcome. There are also little data comparing HF cohorts of differing ethnicity within the Asia-Pacific region.MethodsThe Singapore Heart Failure Outcomes and Phenotypes (SHOP) study and Prospective Evaluation of Outcome in Patients with Heart Failure with Preserved Left Ventricular Ejection Fraction (PEOPLE) study are parallel prospective studies using identical protocols to enroll patients with HF across 6 centers in Singapore and 4 in New Zealand. The objectives are to determine the relative prevalence, characteristics, and outcomes of patients with HF and preserved EF (EF ≥50%) compared with those with HF and reduced EF, and to determine initial data on ethnic differences within and between New Zealand and Singapore. Case subjects (n = 2,500) are patients hospitalized with a primary diagnosis of HF or attending outpatient clinics for management of HF within 6 months of HF decompensation. Control subjects are age- and gender-matched community-based adults without HF from Singapore (n = 1,250) and New Zealand (n = 1,073). All participants undergo detailed clinical assessment, echocardiography, and blood biomarker measurements at baseline, 6 weeks, and 6 months, and are followed over 2 years for death or hospitalization. Substudies include vascular assessment, cardiopulmonary exercise testing, retinal imaging, and cardiac magnetic resonance imaging.ConclusionsThe SHOP and PEOPLE studies are the first prospective multicenter studies defining the epidemiology and interethnic differences among patients with HF in the Asia-Oceanic region, and will provide unique insights into the pathophysiology and outcomes for these patients.     

Apoprotein B100 has a prolonged interaction with the translocon during which its lipidation and translocation change from dependence on the microsomal triglyceride transfer protein to independence

When lipid synthesis is limited in HepG2 cells, apoprotein B100 (apoB100) is not secreted but rapidly degraded by the ubiquitin-proteasome pathway. To investigate apoB100 biosynthesis and secretion further, the physical and functional states of apoB100 destined for either degradation or lipoprotein assembly were studied under conditions in which lipid synthesis, proteasomal activity, and microsomal triglyceride transfer protein (MTP) lipid-transfer activity were varied. Cells were pretreated with a proteasomal inhibitor (which remained with the cells throughout the experiment) and radiolabeled for 15 min. During the chase period, labeled apoB100 remained associated with the microsomes. Furthermore, by crosslinking sec61β to apoB100, we showed that apoB100 remained close to the translocon at the same time apoB100–ubiquitin conjugates could be detected. When lipid synthesis and lipoprotein assembly/secretion were stimulated by adding oleic acid (OA) to the chase medium, apoB100 was deubiquitinated, and its interaction with sec61β was disrupted, signifying completion of translocation concomitant with the formation of lipoprotein particles. MTP participates in apoB100 translocation and lipoprotein assembly. In the presence of OA, when MTP lipid-transfer activity was inhibited at the end of pulse labeling, apoB100 secretion was abolished. In contrast, when the labeled apoB100 was allowed to accumulate in the cell for 60 min before adding OA and the inhibitor, apoB100 lipidation and secretion were no longer impaired. Overall, the data imply that during most of its association with the endoplasmic reticulum, apoB100 is close to or within the translocon and is accessible to both the ubiquitin-proteasome and lipoprotein-assembly pathways. Furthermore, MTP lipid-transfer activity seems to be necessary only for early translocation and lipidation events.     

Inhibition of DNA synthesis by phenobarbital in primary cultures of hepatocytes from normal rat liver and from hepatic nodules.

One of the many hypotheses put forward to explain the mechanism by which phenobarbital (PB) promotes hepatocarcinogenesis is by differential mitoinhibition of surrounding hepatocytes while allowing the initiated hepatocytes to respond to growth stimuli and form foci and nodules. Given the similarity in structures between PB and orotic acid (OA), another rat liver tumor promoter, the present investigation was designed to determine (i) whether PB, like OA, exerts its mitoinhibitory effect at a site beyond the growth factor receptor and receptor mediated early events; and (ii) whether PB exerts a differential mitoinhibitory effect by selectively inhibiting the non-initiated hepatocytes but not the initiated hepatocytes in vitro. Our studies demonstrate that, like OA, PB also inhibits DNA synthesis in hepatocytes from normal rat liver in a dose dependent manner with 80-90% at a dose of 6 mM. One target site may lie beyond the growth factor receptor mediated early events because PB inhibited DNA synthesis in hepatocytes primed with the growth factor 24 h earlier. Interestingly, PB inhibited DNA synthesis not only in hepatocytes from non-nodular surrounding liver but also in hepatocytes from persistent hepatic nodules initiated with 1,2-dimethylhydrazine and promoted with OA. Therefore, our results suggest that although PB is a mitoinhibitor of DNA synthesis in hepatocytes, it does not appear to create as strong a differential mitoinhibition between non-nodular surrounding and initiated hepatocytes as is evident in the resistant hepatocyte and OA models. These results raise the question whether differential mitoinhibition is the major contributing factor in the PB mediated rat liver tumor promotion.