Autoimmune hepatitis in pregnancy: Pearls and pitfalls.

Autoimmune hepatitis (AIH) in pregnancy has many unique considerations. Evidence provided from single center studies with patient level data and nationwide population studies provide valuable insight into this complex situation. Because a planned pregnancy is a safer pregnancy, preconception counseling is a crucial opportunity to optimize care and risk stratify women with AIH. Women with chronic liver disease who receive preconception advice and counseling are more likely to achieve stable liver disease at conception and undergo appropriate variceal surveillance. Loss of biochemical response in pregnancy is associated with adverse outcomes in unstable disease. New onset AIH in pregnancy should be managed with classical treatment regimens. The continued use of immunosuppression in pregnancy, with the exception of mycophenolate mofetil, has not shown to adversely affect the rates of stillbirth or congenital malformation. Previously adopted immunosuppression withdrawal paradigms in pregnancy should no longer be considered advantageous, because remission loss postdelivery is likely (12%-86%). Population studies, report improved outcomes with preterm birth rates falling from 20% to 9%-13% in AIH pregnancies over a 20-year period. Newer data have also demonstrated an increased risk of gestational diabetes and hypertensive complications in AIH pregnancy, which has implications for management and preeclampsia prevention with aspirin use. This review aims to provide the framework to guide and manage pregnancy in AIH outlining pearls and pitfalls to ensure optimal outcomes for mother, baby and to reduce variation in practice.

Acute stroke patients with high BMI are less likely to have severe disability at initial presentation.

This is a prospective study to determine the severity of disability and prognosis of acute stroke patients related to their Body Mass index (BMI). A total of 79 consecutive CT-scan-proven acute stroke patients who were admitted to Hospital Tuanku Ja'afar, Seremban between November 2006 and April 2007 were recruited (male:female 49:30; mean age 62.7 years; ischemic stroke 70, intracerebral bleed 9). The patients were divided according to BMI less than 25 (Group A) and equal or greater than 25 (Group B). Severity of disability was measured between 24-48 hours by modified Rankin's score. Patients were followed up after one month. Thirty-seven patients had severe disability (Rankin Score 5). Twenty-nine patients had adverse outcomes including 11 deaths and 18 rehospitalizations or prolonged hospital/nursing home stay. 34.3% of Group B had severe disability compared to 56.8% of Group A (chi2 P = 0.046). Conversely 42.9% of Group B had adverse events at one month compared to 31.8% of Group A (chi2 P = 0.312). There were no statistical differences between high- and low-BMI groups for gender ratio, smoking, hypertension, diabetes, prior cardiovascular disease, mean age, mean lipid profile and blood pressure. When comparing patients with Rankin Score 1-4 versus 5, age and BMI were statistically significant between the two groups. By multivariate analysis only age is independent predictor for severe disability (P < 0.05). The results of this pilot study should be confirmed in larger prospective multicentre trial.

Bone ingrowth on a smooth-surfaced hydroxyapatite-coated acetabular cup.

We studied 29 retrieved smooth-surfaced hydroxyapatite (HA)-coated acetabular cups that had been in situ for an average of 54.9 months. The outer surface of all cups carried two circular grooves dividing the smooth surface. The extent of bony in-growth and hydroxyapatite absorption was calculated using a computer imaging system (analySIS-pro 3.0, Soft Imaging System GmbH, Germany). Absorption of HA was seen in all 29 cups. The extent of absorption was, on average, 60.5%. Twenty-five cups showed bony in-growth covering an average of 13.8% of the cup surface. Thirteen cups showed bony in-growth on the smooth surface as well as the circular grooves. On 11 cups, the bony in-growth was confined to the grooves. We found that bony in-growth on the smooth-surfaced HA-coated acetabular cup was minimal. It occurred along the two circular grooves rather than on the smooth surfaces. We conclude that a macro-structure surface enhancing bony in-growth is necessary for the long-term survival of the studied cup.

Controlling self-incompatibility by CO2 gas treatment in Brassica campestris: structural alteration of papillae cell and differential gene expression by increased CO2 gas.

Self-imcompatibility is a controlling genetic mechanism to prevent self-pollination for Chinese cabbage (Brassica campestris), one of the major vegetable crops in Korea. To maintain inbred lines of the crop plant, a method in that high CO2 gas is treated to the pistils to overcome the self-incompatibility and thereby self-pollens can successfully make germination and fertilization has been widely used in seed companies. Despite the common utilization of this method, any molecular and cellular studies on how the self-incompatibility is removed from the Chinese cabbage plant have not been done. In this study, we show that the increased CO2 gas causes a structural alteration of the papillae cell and thereby the self-incompatibility is removed from the Chinese cabbage plant, allowing the self-pollens to germinate and penetrate the papillae cell. Also, gene expression in the pistil treated with CO2 gas was studied by DD/RT-PCR and reverse northern hybridization experiments. The results suggest that the failure in self-incompatible reaction resulted not only from the structural alteration of the papillae cell but also from change in the pistil component production that is either positively or negatively regulated by the environmental stimulation.

Cytological observation of two environmental genic male-sterile lines of rice.

We report here two environmental genic male-sterilities (EGMS) in rice. These two EGMS rice lines, thermo-sensitive genic male-sterility (TGMS) and photoperiod-sensitive genic male-sterility (PGMS), are controlled by temperature and photoperiod, respectively, in determining their male-sterility. Male-sterility of the TGMS and PGMS was found to be induced when they were grown at 32 degrees C/26 degrees C (day/night) with 14 h daylight, while they were fertile at 26 degrees C/20 degrees C (day/night) with 10 h daylight in a growth chamber. We also examined their anther structures under a light microscope. The light microscopic observation revealed that the EGMS lines showed a complete pollen abortion at the sterile growth condition while they produced normal fertile pollens at the fertile growth condition.

Postnatal development of parvalbumin and calbindin D-28k immunoreactivities in the canine hippocampus.

The calcium-binding proteins, parvalbumin and calbindin D-28k, are markers of different classes of GABAergic interneurons and display different functions. The present study was attempted to determine immunoreactivities and colocalization of the parvalbumin and calbindin D-28k in the developing canine hippocampus by immunohistochemistry. The calcium-binding protein-containing neurons showed different developmental patterns. The first appearance of parvalbumin immunoreactive nonpyramidal cells was observed at P7. Parvalbumin immunoreactivity was elicited by the sequence from CA3 to CA1 to reach an adult-like distribution pattern, which was reached at P60, while calbindin D-28k immunoreactivity appeared from P0, including pyramidal and nonpyramidal cells. The characteristic distribution of calbindin D-28k immunoreactive pyramidal cells was clarified by P28, and an adult-like distribution pattern was reached by the end of the second postnatal month. Double-labeled nonpyramidal cells were frequently seen in the subareas, CA3 of P14/CA1-CA2 of P28, where parvalbumin immunoreactive nonpyramidal cells were emerging. These data suggest that the colocalization of the two calcium-binding proteins during development is related closely to the area-specific maturation of parvalbumin expression, although either prenatal expression of calbindin D-28k or parvalbumin was not determined.

Single-stranded endonuclease activity in the excretory--secretory products of Trichinella spiralis and Trichinella pseudospiralis.

A novel acidic extracellular single-stranded endonuclease was demonstrated for the first time in the excretory-secretory (E-S) products of 2 species of Trichinella. Unlike the double-stranded endonuclease reported earlier, the single-stranded molecule is divalent cation independent and is detected in both T. spiralis and T. pseudospiralis E-S products. It hydrolysed single-stranded DNA and RNA at comparable rates. The single-stranded endonuclease was sensitive to inhibition by Zn2+ and to high concentrations of NaCl. Zymographic analysis indicated that it was encoded by at least 3 peptides of Mr approximately 50-60 kDa. The rate of hydrolysis of single-stranded targets by the E-S products was substantially higher than that of the double-stranded molecule. Due to the differences in peptide profile, divalent cation dependence, and species-specific expression, the single and double-stranded endonucleases are likely to be encoded by different proteins and may have different functions.

leafy hull sterile1 is a homeotic mutation in a rice MADS box gene affecting rice flower development.

Rice contains several MADS box genes. It has been demonstrated previously that one of these genes, OsMADS1 (for Oryza sativa MADS box gene1), is expressed preferentially in flowers and causes early flowering when ectopically expressed in tobacco plants. In this study, we demonstrated that ectopic expression of OsMADS1 in rice also results in early flowering. To further investigate the role of OsMADS1 during rice flower development, we generated transgenic rice plants expressing altered OsMADS1 genes that contain missense mutations in the MADS domain. There was no visible alteration in the transgenic plants during the vegetative stage. However, transgenic panicles typically exhibited phenotypic alterations, including spikelets consisting of elongated leafy paleae and lemmas that exhibit a feature of open hull, two pairs of leafy palea-like and lemma-like lodicules, a decrease in stamen number, and an increase in the number of carpels. In addition, some spikelets generated an additional floret from the same rachilla. These characteristics are very similar to those of leafy hull sterile1 (lhs1). The map position of OsMADS1 is closely linked to that of lhs1 on chromosome 3. Examination of lhs1 revealed that it contains two missense mutations in the OsMADS1 MADS domain. A genetic complementation experiment showed that the 11.9-kb genomic DNA fragment containing the wild-type OsMADS1 gene rescued the mutant phenotypes. In addition, ectopic expression of the OsMADS1 gene isolated from the lhs1 line resulted in lhs1-conferred phenotypes. These lines of evidence demonstrate that OsMADS1 is the lhs1 gene.

Isolation and characterization of a class I SLG gene from Chinese cabbage (Brassica campestris).

In Brassica species, self-incompatibility in the recognition reaction between self and non-self pollens is determined by two genes, SLG and SRK, at the S locus. We have cloned and characterized a genomic DNA fragment containing a complete open reading frame of the SLG gene from Chinese cabbage. The genomic clone, named BcSLG2, was found to possess the region that shares a homology of 77% in amino acid identity with the SLG46 gene of Brassica campestris. Northern blot analysis revealed that the BcSLG2 gene expression is restricted to the pistil of Chinese cabbage flower. In situ hybridization showed that in the pistil, the gene is expressed predominantly in the stigmatic tissue. Much lower expression in the tapetum was also detectable at an immature stage of the flower development. Southern blot hybridization with the BcSLG2 DNA probe showed polymorphism in the SLG gene organization of the Chinese cabbage plants. These results will provide valuable information in understanding the S gene complex of the Chinese cabbage plants.

The distribution of excretory/secretory antigens during the muscle phase of Trichinella spiralis and T. pseudospiralis infections.

The in situ distribution of excretory/secretory (ES) antigens of the infective-stage larvae of Trichinella spiralis and T. pseudospiralis was compared at various periods of development by immunofluorescent laser confocal microscopy and the immunoperoxidase method. In the former infection, epitopes of the ES antigens were always confined exclusively within the nurse cell, i.e., in the cytoplasmic region, hypertrophic nuclei, stichocytes, and cuticular surface of worms. In the latter infection, as early as at day 15 postinfection, ES epitopes were located along the infected myofibers, in the adjacent muscles, hypertrophic nuclei, stichocytes, and cuticular surface of worms. By day 30 postinfection there was a marked increase in both the distribution and the intensity of ES antigens in infected as opposed to uninfected myofibers. A new method was also developed to reveal the number of hypertrophic nuclei, small cells, and larvae in intact nurse cells. As many as four worms could be accommodated within a single complex. The number of hypertrophic nuclei within each complex varied from 15 to 81.

Cloning and characterization of the chloroplast elongation factor EF-Tu cDNA of Oryza sativa L.

From the rice leaf cDNA library, we have cloned a cDNA encoding rice chloroplast translational elongation factor EF-Tu (tufA). The rice tufA cDNA clone contains 1678 nucleotides and codes for a 467 amino acid protein including a putative chloroplast transit peptide of 59 amino acid residues. The predicted molecular mass of the mature protein is approximately 45 kDa. This cDNA clone contains the 61 nucleotides of the 5' untranslated region (UTR) and the 213 nucleotides of 3' UTR. Amino acid sequence identity of the rice tufA with the mature chloroplast EF-Tu proteins of tobacco, pea, arabidopsis, and soybean ranges from 83% to 86%. The deduced polypeptide of the rice tufA cDNA contains GTP binding domains in its N-terminal region and chloroplast EF-Tu signature regions in the C-terminal region. The rice tufA appears to exist as a single copy gene, although its homologues of maize and oat exist as multiple copy genes. The rice tufA gene is located in chromosome 1 and is more highly expressed in the leaf than in root tissue.

A novel cDNA clone encoding a specific excretory/secretory antigen of larval Trichinella pseudospiralis.

A cDNA library for Trichinella pseudospiralis was constructed to study the expression of specific antigens. Four positive clones were identified using antibodies against the excretory/secretory (ES) products of the nematode as probe. Sequence analysis showed that they contained identical cDNA inserts of 606 bp, including a 5' non-translated region of 96 bp, a core translated segment of 408 bp and a poly(A)+ 3' terminus. It encoded a novel 136-amino-acid polypeptide. Southern blot analysis indicated that the cDNA did not cross-hybridize to the genomic digests of T. spiralis, mouse, or rat. A single copy only of its complementary sequence was found in the genome of T. pseudospiralis. Using the lambda ZAP expression system, the cDNA was induced to express a 23-kDa beta-galactosidase-fusion protein which did not cross-react with polyclonal and monoclonal antibodies against T. spiralis, heat shock proteins, or four heterologous species of nematodes. The antiserum against the fusion protein recognized a 15-kDa band from the ES products of T. pseudospiralis in immunoblotting. Immunocytolocalization demonstrated that the anti-fusion protein serum only recognized an epitope in the stichosome of T. pseudospiralis and not in T. spiralis. The protein can therefore serve as a specific antigen for the differential diagnosis of trichinellosis.

Isolation and characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.).

An anther-specific cDNA clone of rice, RA8, was isolated from an anther cDNA library by differential screening. RNA blot analysis indicated that the RA8 transcript is present specifically in anthers and the transcript level increased as flowers matured, reaching the highest level in mature flowers. The RA8 clone contains an open reading frame of 264 amino acid residues with a hydrophobic N-terminal region. The deduced amino acid sequences did not show significant homology to any known sequences. Genomic DNA blot analysis showed that RA8 is a single-copy gene. A genomic clone corresponding to the RA8 cDNA was isolated and its promoter region was fused to the beta-glucuronidase (GUS) gene. Transgenic rice plants exhibited anther-specific expression of the GUS reporter gene. Histochemical GUS analysis showed that the RA8 promoter was active in the tapetum, endothecium, and connective tissues of anthers. Experiments showed that expression of the gene starts when microspores are released from tetrads, and it reaches to the maximum level at the late vacuolated-pollen stage. The RA8 promoter may be useful for controlling gene expression in anthers of cereal plants and for generating male-sterile plants.

Ectopic expression of tobacco MADS genes modulates flowering time and plant architecture.

MADS genes encode regulatory factors that are involved in various developmental steps of the plant life cycle. During flower development, they regulate the early step of specifying floral meristem identity as well as the later step of determining the fate of floral organ primordia. Here, we report the isolation of two cDNA clones, NsMADS2 and NsMADS3, from a long-day tobacco species, Nicotiana sylvestris, which encode MADS domain-containing proteins. The NsMADS2 amino acid sequence showed 66% identity to SQUA and 64% to AP1. NsMADS3 showed a high degree of amino acid identity with FBP2 (98%), DEFH 72 (89%), DEFH 200 (88%), and AGL9 (76%). RNA blot analyses of NsMADS2 and NsMADS3 revealed that both transcripts were present in floral organs, but not in vegetative organs such as the leaf, root, stem, and 10 d old seedlings. The NsMADS2 transcript was localized in all four whorls and the NsMADS3 transcript was restricted in the three inner whorls of floral organs. The ectopic expression of NsMADS2 using the CaMV 35S promoter caused early flowering and lengthened internode length in transgenic tobacco plants. The ectopic expression of NsMADS3 also caused early flowering phenotype in transgenic tobacco plants, but the plants exhibited reduced apical dominance. Possible implications of these results in relation to the functions of NsMADS2 and NsMADS3 are discussed.

Surgical closure of atrial septal defect. Minimally invasive cardiac surgery or median sternotomy?

BACKGROUND: Closure of ostium secundum atrial septal defect (ASD) vis median sternotomy (MS) is a simple procedure for most cardiac surgeons. Minimally invasive cardiac surgery (MICS) has recently been applied in the management of intracardiac lesions. METHODS: We report our experience in surgical closure of isolated ASD via MICS in 60 patients and via MS in 58 patients. There was no difference between these two groups in gender, age, body weight, ratio of systemic to pulmonary blood flow, and pulmonary arterial pressure. RESULTS: The duration of cardiopulmonary bypass was significantly longer in the MICS group than in the MS group [27 to 126 min (42 +/- 12) and 14 to 158 min (27 +/- 11), respectively; (p < 0.001]. However, the length of incision, incidence of temporary pacemaker wire insertion rate, duration of endotracheal intubation, timing of oral intake, postoperative day drainage amount, incidence of parenteral analgesic injection, postoperative length of stay, and return to normal activity interval were significant shorter and lower in patients of the MICS group than in those of the MS group. All the patients recovered rapidly from the surgery. Follow-up was complete in all patients, with no late complications and no residual shunt. CONCLUSION: Our results suggest that MICS is a good option for surgical closure of ASD.

Characterization of a tobacco MADS-box gene homologous to AGL2.

Flower development is regulated by a group of regulatory factors containing the MADS box domain. Recently, we isolated and characterized a tobacco MADS-box gene, NtMADS1, which is homologous to AGL2 in Arabidopsis. Sequence analysis revealed that NtMADS1 encodes a putative MADS-box protein homologous to the AGL2 with 68.4% identity and 82% similarity in amino acids. The NtMADS1 gene started to express at a very early stage of flower development, such as floral primodia, and the expression was maintained during the entire floral development. In the tobacco flower, the gene was expressed in all four floral organs, namely the sepal, petal, anther, and carpel, and was not detected in the vegetative organs such as the leaf and root. In situ hybridization was also performed to monitor the detailed expression pattern of NtMADS1 at the tissue level. These results indicate that NtMADS1 is the AGL2 homolog of tobacco and plays important roles in the floral development.

A case of membranous glomerulonephritis associated with gastric cancer.

We describe a patient with gastric cancer and membranous glomerulonephritis (MGN). The patient, a 61-year-old male, was admitted to our Hospital in May, 1996, because of proteinuria and hyperlipidemia persisting for a year. Laboratory examination filled the criteria of nephrotic syndrome and renal biopsy revealed MGN of stage II. Prednisolone therapy (40 mg/day p.o.) was started, followed by a gradual decrease in proteinuria from 4.5 g/day to 0.1 g/day. Endoscopic examination was performed because of stomach-ache revealed advanced gastric cancer of Borrmann 4. Desiring for a conservative therapy, he was discharged and moved to a hospice. In literature review, MGN is the most frequent lesion among various glomerular diseases associated with malignancy, such as the lung, stomach, and colon, particularly at an elderly ages, and sometimes antedates the detection of malignancy, as in the present case. In several cases with MGN, immune-complexes composed of tumor antigens, such as carcino-embryonic antigen, and antibodies have been reported to deposit in basement membrane of glomeruli, causing MGN. In the renal and gastric cancer tissues of the present case, the presence of three novel tumor-associated antigens, Span-1, Thomsen-Friedenreich antigen (T antigen) and F1 alpha antigen, was examined, using a immuno-peroxidase method. Although none of these three antigens were immuno-stained in the renal tissue, clinical course and literature review suggest that MGN in this patient seems to be associated with gastric cancer, which may have produced MGN-causing tumor antigens other than the three antigens. It should be emphasized that malignancy should be carefully and routinely examined in patients with MGN, particularly at elderly ages.