Association of sociodemographic and maternal healthcare factors with birth registration in Angola.

OBJECTIVES: Angola has a high burden of unregistered children and efforts to increase birth-registration coverage have not yielded the desired progress. This study aimed to examine sociodemographic and healthcare-related factors associated with birth registration in Angola. STUDY DESIGN: Secondary data analysis of the Maternal and Child Health (MCH) Handbook randomised controlled trial conducted in Benguela province, Angola and involving 11,006 women. METHODS: For this analysis, we excluded women with missing data on birth registration (n = 1424), multiple gestation (n = 243), and those with infant death (n = 6). The final study population included 9333 women with infants under one year of age. We used multilevel mixed-effects logistic regression analysis to determine sociodemographic and healthcare-related factors associated with the registration of a child's birth. RESULTS: Of the 9333 live births, 25% (95% confidence interval [CI] = 13.4-41.8) were registered, while 21% (95%CI = 11.1-35.7) were registered with certificate. There were higher proportions of registered births among mothers who possessed the MCH Handbook across various demographic and healthcare indicators. Birth registration was most significantly associated with facility-based delivery (odds ratio [OR] = 2.97; 95%CI = 2.45-3.61), possession of MCH Handbook (OR = 2.04; 95%CI = 1.70-2.46), and complete scheduled vaccination visits (OR = 1.69; 95%CI = 1.44-1.97). Higher maternal age and education level, belonging to the highest wealth quintile, beginning antenatal care in the first trimester, attending at least four antenatal care visits, and using postnatal care services were positively associated with registration of birth. CONCLUSION: Maternal healthcare factors showed significant associations with birth registration and integrating birth-registration processes with certain maternal and child health services may further raise awareness and boost registration levels in Angola.

Observation and determination of periodontal tissue profile using optical coherence tomography.

BACKGROUND AND OBJECTIVE: Diagnosis is a crucial step in periodontal treatment. The aim of this study was to evaluate the effectiveness of optical coherence tomography (OCT) for observation and determination of periodontal tissue profiles in vivo. MATERIAL AND METHODS: In experiment 1, refractive indices of purified water, porcine gingiva and human gingiva at 1330 nm were determined for the analysis of OCT images of periodontal tissues. In experiment 2, OCT examination was performed in the midlabial apico-coronal plane of mandibular anteriors in 30 Asian volunteers with healthy gingiva. Sulcus depth was measured on intra-oral photographs taken during probing. In the OCT images, the gingival, epithelial and connective tissue thickness, and the position of alveolar bone crest were determined and finally, the biologic width was measured. RESULTS: Refractive indices of purified water, porcine gingiva and human gingiva were 1.335, 1.393 and 1.397, respectively. Cross-sectional images of gingival epithelium, connective tissue and alveolar bone were depicted in real-time. The sulcular and junctional epithelium could be visualized occasionally. Laser penetration and reflection were limited to a certain depth with an approximate maximal imaging depth capability of 1.5 mm and OCT images of the periodontal structure were not clear in some cases. The average maximal thickness of gingiva and epithelium and biologic width at the mandibular anteriors were 1.06 ± 0.21, 0.49 ± 0.15 and 2.09 ± 0.60 mm, respectively. CONCLUSION: OCT has promise for non-invasive observation of the periodontal tissue profile in detail and measurement of internal periodontal structures including biologic width in the anterior region.

Factors Affecting Steady-state Plasma Concentrations of Enantiomeric Mirtazapine and its Desmethylated Metabolites in Japanese Psychiatric Patients.

INTRODUCTION: This study evaluated the effects of the CYP2D6*10 genotype on steady-state plasma concentrations of enantiomeric mirtazapine (MIR) and N-desmethylmirtazapine (DMIR) in Japanese patients. METHODS: Subjects were 77 Japanese patients treated with racemic MIR. Steady-state plasma concentrations of MIR and DMIR enantiomers were measured using stereoselective liquid chromatography. Polymerase chain reaction was used to determine the CYP2D6 genotypes. RESULTS: After correcting for dose and body weight, smokers (n=15) had significantly lower S-(+)-MIR than nonsmokers (n=55) (15.1±17.8 vs. 23.9±17.8 ng/mL/mg/kg, Kruskal-Wallis test, p=0.034). One-way analysis of variance revealed that CYP2D6*10 homozygotes had significantly higher corrected plasma concentrations of S-(+)-MIR than the no-variant allele group (p=0.034). Multiple regression analysis revealed a significant positive correlation between the number of CYP2D6*10 alleles and corrected plasma concentrations of S-(+)-MIR. These results yielded the following final model: corrected plasma concentration of S-(+)-MIR=15.9+7.30×(number of CYP2D6*10 alleles) (R=0.279, p=0.023, coefficient of determination (R(2))=0.078). CONCLUSION: Homozygous CYP2D6*10 alleles and smoking have a significant impact on the metabolism of S-(+)-MIR in Japanese patients.

Antimicrobial effect of photodynamic therapy using high-power blue light-emitting diode and red-dye agent on Porphyromonas gingivalis.

BACKGROUND AND OBJECTIVE: Antimicrobial photodynamic therapy (a-PDT) using a combination of red-colored laser/light-emitting diode (LED) and blue dye has been employed for periodontal therapy and the antimicrobial effect seems promising. Blue light, which has favorable wavelength properties, would be more effective as a light source for a-PDT because blue light itself possesses an antimicrobial effect. This study aimed to investigate the effect of a-PDT using a novel combination of high-power blue LED and red-dye agent on Porphyromonas gingivalis in vitro. MATERIAL AND METHODS: Porphyromonas gingivalis ATCC 33277 suspension was irradiated with blue LED (BL) (425-470 nm) or red LED (RL) (625-635 nm) at 30-90 J/cm(2) , or was mixed with erythrosine (ER), phloxine B (PB) or rose bengal (RB) with or without BL irradiation (30 J/cm(2) ). RL (30 J/cm(2) ) in combination with toluidine blue was employed as positive control. All the suspensions of P. gingivalis were serially diluted, plated and incubated anaerobically, and the numbers of colony-forming units (CFUs) were counted on day 7. RESULTS: BL irradiation at 60 and 90 J/cm(2) demonstrated a significant reduction in the numbers of CFUs. ER, PB and RB solutions at 160 µg/mL showed almost no or only a minimal reduction in the numbers of CFUs. BL at 30 J/cm(2) combined with ER, PB or RB at 160 µg/mL resulted in a log reduction of 0.9, 1.0 and 7.1, respectively, in the numbers of CFUs; 30 J/cm(2) BL with RB at 1.6, 16 and 160 µg/mL demonstrated a log reduction of 6.3, 8.0 and 5.5, respectively; and a log reduction of 5.2 was obtained after 30 J/cm(2) RL with 16 µg/mL TB. CONCLUSION: Within the limits of this study, BL was found to have an antimicrobial/growth-inhibiting effect on P. gingivalis, and a-PDT using a combination of BL and RB shows promise as a new technical modality for bacterial elimination in periodontal therapy.

[Resection of a right ventricular diverticulum of fibrous type by off-pump cardiac surgery; report of a case].

Right ventricular diverticulum is very rare and we experienced a case of isolated right ventricular diverticulum in an adult patient The patient was an 80-year-old man and a 3-cm-diameter round mass at the apex of the heart was pointed out by screening computed tomography (CT). A small and akinetic diverticulum having a narrow communication with the right ventricle was revealed by right ventriculography. Upon surgery, a 3-cm-diameter diverticulum was found at the acute margin of the right ventricle. The diverticulum was exposed using the off-pump coronary artery bypass grafting (CABG) technique. Two mattress sutures of 4-0 Prolene with Teflon felt strips were used to close the communication between the diverticulum and the right ventricle, then, the diverticulum was resected. His postoperative course was uneventful. Pathological examination revealed the endothelial-lined wall of the diverticulum consisting of internal elastic lamina and discontinuous thin smooth muscle layer with no myocardium. This type of right ventricular diverticulum could be resected by the off-pump CABG technique.

Enzymatic properties of de novo-type mouse DNA (cytosine-5) methyltransferases.

We have purified GST-fused recombinant mouse Dnmt3a and three isoforms of mouse Dnmt3b to near homogeneity. Dnmt3b3, an isoform of Dnmt3b, did not have DNA methylation activity. Dnmt3a, Dnmt3b1 or Dnmt3b2 showed similar activity toward poly(dG-dC)-poly(dG-dC) for measuring de novo methylation activity, and toward poly(dI-dC)-poly(dI-dC) for measuring total activity. This indicates that the enzymes are de novo-type DNA methyltransferases. The enzyme activity was inhibited by NaCl or KCl at concentrations >100 mM. The kinetic parameter, K(m)(AdoMet), for Dnmt3a, Dnmt3b1 and Dnmt3b2 was 0.4, 1.2 and 0.9 microM when poly(dI-dC)-poly(dI-dC) was used, and 0.3, 1.2 and 0.8 microM when poly(dG-dC)-poly(dG-dC) was used, respectively. The K(m)(DNA) values for Dnmt3a, Dnmt3b1 and Dnmt3b2 were 2.7, 1.3 and 1.5 microM when poly(dI-dC)-poly(dI-dC) was used, and 3.5, 1.0 and 0.9 microM when poly(dG-dC)-poly(dG-dC) was used, respectively. For the methylation specificity, Dnmt3a significantly methylated CpG >> CpA. On the other hand, Dnmt3b1 methylated CpG > CpT >/= CpA. Immuno-purified Dnmt3a, Myc-tagged and overexpressed in HEK 293T cells, methylated CpG >> CpA > CpT. Neither Dnmt3a nor Dnmt3b1 methylated the first cytosine of CpC.

Effect of hyperthermia on both primary proliferation and self-renewal of human leukemic progenitor cells in vitro: its application to in vitro purging.

Self-renewal, as defined by the capacity to yield new colonies following replating, is an important function of leukemic progenitors (L-CFU) to originate self-maintaining clones. In this work, we studied the effect of hyperthermia (41-44 degrees C) on the growth of human L-CFU derived primary colonies and of secondary colonies formed by replating to evaluate the purging effect of human L-CFU by heat. The survival curves clearly demonstrated much greater hyperthermic sensitivity of L-CFU compared to normal granulocyte-macrophage progenitors (CFU-GM) at all temperatures (41-44 degrees C) studied. At 42 degrees C or higher, L-CFU decreased (by more than 2 log reduction) dramatically and therefore were unable to form colonies in vitro. At 42 degrees C and 43 degrees C, 65 and 30%, respectively, of CFU-GM obtained from remission and normal marrows were left after 1 h exposure. At 44 degrees C, however, CFU-GM derived colonies disappeared after a 2 h exposure. In the four available patients with acute myelogenous leukemia, secondary colonies formed by replating were decreased in proportion to the decreasing primary colonies during heating (42 and 43 degrees C). However, their self-renewal capacity was retained in vitro until the primary colonies disappeared. These observations suggest that heat exposure at 43 degrees C for 1 h could be the most effective conditions for in vitro purging of human L-CFU because of the wide difference between surviving fractions of CFU-GM and L-CFU.

[Nonpenetrating traumatic injury of the thoracic aorta and left subclavian artery; report of a case].

Nonpenetrating traumatic injury of the thoracic aorta and/or its major branch is usually fatal and the treatment of this condition carries extremely high risk because of associated visceral organ injuries. Accurate diagnosis have been difficult. However, recently developed multi-slice helical computed tomography (CT) is highly sensitive in early detection of precise location of injury and associating injuries of other organs. Here we report our case with combined thoracic aortic and left subclavian artery injuries, diagnosed by 3-dimensional (3-D) CT and treated successfully.

Efficient introduction of a gene into hematopoietic cells in S-phase by electroporation.

Cells of the hematopoietic cell line K562 were synchronized by three different methods: single aphidicolin treatment, thymidine treatment followed by hydroxyurea exposure, and double hydroxyurea treatment. The synchronized cells were transfected via electroporation with plasmid pMoZtk, which contains the beta-galactosidase gene, using a square wave pulse immediately after synchronization or at various time points during culture. Simultaneously, synchronized cells were fluorescence-activated cell sorter (FACS) analyzed to determine their stage in the cell cycle using double staining with bromodeoxyuridine (BrdU) and propidium iodide. Highly efficient introduction of pMoZtk was observed for the cell fraction, which predominantly consisted of the cells in S-phase. These results suggest that by increasing the proportion of cells in S-phase, the efficiency of gene transfer into hematopoietic cells such as hematopoietic stem cells can be improved.

Differentiation of endocrine myocardiocytes in the developing heart of the toad (Bufo arenarum Hensel).

The differentiation of endocrine myocardiocytes was investigated in the heart of developing toad Bufo arenarum Hensel, combining ultrastructural and immunocytochemical procedures. The distribution of immuno-reactive atrial natriuretic peptide (ANP) in the whole heart was appraised by light microscopy, applying biotin-streptavidin and immunofluorescence techniques. With the latter procedures ANP was first recognized at embryonic stage 22, in both atrium and ventricle. In the ensuing stages the ANP-reactivity became stronger in the atrium, while it became dimmer in the ventricle. At the end of the larval prometamorphic stage, atrial myocardiocytes acquired almost all the features of adult myoendocrine cells. At electron microscope level, small inclusions, about 110-120 nm in diameter, resembling secretory granules were found in myoendocrine cells beginning at embryonic stage 22. However, no immunogold labeling of ANP occurred until stage 25. The number of secretory granules diminished in the ventricles and increased in the atrium of the larval heart and at the end of the prometamorphic stage the atrial myoendocrine cells presented the ultrastructural characteristics of active secretory cells. The synthesis of ANP in larvae is enhanced at a critical period of development when the developing toad switches from an aquatic environment to terrestrial life. The cardiac hormones seem to play a key role in the regulation of the osmolarity of body fluids at this developmental stage.

Lack of association between the Trp64 Arg mutation in the beta 3-adrenergic receptor gene and obesity in Japanese men: a longitudinal analysis.

The beta 3-adrenergic receptor (beta 3AR) is implicated in the regulation of thermogenesis and lipolysis, and it is suggested that the Trp64 Arg mutation in this receptor may contribute to the development of obesity. To examine whether the Trp64 Arg mutation had any effect on body weight during adult life, the beta 3AR genotype was determined in 186 unselected Japanese men, most of whom had records of body weight measured yearly from 25-53 yr of age. Of them, 26 subjects were diagnosed as having noninsulin-dependent diabetes mellitus (NIDDM) and 41 as having impaired glucose tolerance. There were 6 subjects (3%) with homozygous mutation, 67 (36%) with heterozygous mutation, and 113 (61%) with normal allele. Among the 3 genotypes, there were no significant differences in body mass index (BMI) at any age between 25-53 yr and the prevalence of NIDDM at the age of 53 yr. When longitudinal changes in body weight were compared between subjects with and without mutation, the former were less prone to gain weight than the latter. The frequency of the mutant allele was 1) not different among obese (BMI, > 26.4), intermediate (BMI, 22-26.4), and nonobese (BMI, < 22.0) subjects (0.21, 0.22, and 0.26, respectively; P = 0.77); 2) lower in subjects with NIDDM than in those without it, but the difference was insignificant (0.12 vs. 0.23; P = 0.07); and 3) similar between 186 unselected men and another group of 100 patients with NIDDM that were randomly selected for comparison (0.21 vs. 0.23). These results suggest that the beta 3AR is not a major contributing factor to obesity or NIDDM in Japanese men.

Effects of cholesterol and apolipoprotein E on retinal abnormalities in ApoE-deficient mice.

PURPOSE: To examine the pathologic changes in the retina of apolipoprotein E (apoE)-deficient mice fed a high-cholesterol diet. METHODS: ApoE-deficient mice (ApoE) were maintained on either regular mouse chow (ApoE-R) or a high-cholesterol diet (ApoE-C) for 25 weeks. Age-matched control C57BL/6J mice (C57) were also maintained on either regular mouse chow (C57-R) or a cholesterol-containing diet (C57-C). Retinal function was assessed by dark-adapted electroretinography (ERG). The eyes were embedded, sectioned, and analyzed by histologic and immunohistochemical methods, as well as by light and transmission electron microscopy. RESULTS: After the 25-week feeding period, ERG tracings of ApoE-C mice revealed significant increases of a- and b-wave implicit times when compared with the C57-R group of mice. In addition, there were reductions in oscillatory potential (OP) amplitudes in the ApoE-C group. However, a- and b-wave amplitudes appeared to be unchanged among the four groups of mice. Light microscopic examination of the retinas showed that compared with control C57-R mice, ApoE-C mice had significantly lower cell numbers in the inner and outer nuclear layers (85.1% +/- 4.6%, P < 0.05 and 81.4% +/- 3.7%, P < 0.01 of C57-R controls, respectively). Transmission electron microscopy of apoE-deficient mice revealed cells of the inner nuclear layer with condensation of nuclear chromatin and perinuclear vacuolization in focal areas. Bruch's membrane was also found to be thicker, and its elastic lamina appeared disorganized and discontinuous. Immunohistochemistry demonstrated diminished or no immunoreactivity for carbonic anhydrase II and calretinin in the retinal layers of apoE-deficient mice. CONCLUSIONS: Overall, there were increasing abnormalities of retinal function and cellular morphology among the four groups of mice in the order of C57-R < C57-C < ApoE-R < ApoE-C. These findings suggest that apoE and/or cholesterol play an important role in retinal function.

Release of big and small molecular forms of prolactin: dependence upon dynamic state of the lactotroph.

Secretion of different molecular forms of prolactin was studied from a multidisciplinary standpoint in three different experimental rat models which covered the broadest range of lactotrophic activity. The results obtained allowed the recognition of common secretory patterns in stimulated, inhibited and hyperstimulated lactotrophic activity. Polymeric (big) prolactin is stored exclusively in membrane-bound secretory granules and appears to be a convenient biochemical marker with which to quantify the pool of hormone stored in the cell cytoplasm, while monomeric (small) prolactin represents a pool of newly synthesized hormone which is loosely coupled to organelles involved in its intracellular processing. Most of the prolactin in stimulated lactotrophs is processed through a regulated pathway and the hormone released by exocytosis of secretory granules. Interruption of stimulation resulted in accumulation of secretory granules and polymeric prolactin. In hyperstimulated lactotrophs the polymerization and aggregation of prolactin into secretory granules was bypassed and most of the hormone released directly after its synthesis. The pattern of prolactin secretion could be closely correlated with the dynamic state of the lactotroph.

Subcellular compartmentation of prolactin in rat lactotrophs.

The intracellular localization of different molecular forms of prolactin was studied in various experimental models covering a wide range of secretory states. By correlating electron microscopy, morphometry and quantification of monomeric (small) and polymeric (big) prolactin after differential extraction procedures, big prolactin was found stored in secretory granules while small prolactin was loosely associated with all organelles involved in hormone synthesis and processing. No correlation with levels of lactotrophic secretory activity was detected by either the number of secretory granules or prolactin content in lactotrophs.

Influence of lactotroph cell density on prolactin secretion in rats.

The relationships between the stimulation of prolactin secretion and proliferation of lactotrophs was studied from a multidisciplinary standpoint in three experimental models. Administration of both oestrogen and sulpiride resulted in a significant increase in prolactin secretion and in the lactotroph population. A single injection of 10 micrograms oestradiol benzoate (OB) induced a twofold increase in the proliferation of lactotrophs (morphometrically as volume density), which increased further (2.5-fold) after three OB injections. Parallel changes were observed in the net counts made on lactotrophs sectioned through the nucleus to avoid possible distortions in volume density caused by hypertrophic cytoplasms. Comparable results were obtained with the mitotic index in the same groups of rats exposed to treatment with colchicine. The effect of sulpiride on proliferation of lactotrophs was also significant (1.7-fold) but less pronounced than in rats treated with oestrogens. The treatments with oestrogen and sulpiride did not stimulate lactotrophic activity in a similar way, as judged by the levels of serum prolactin and the storage patterns of small and big prolactin in pituitary glands. Serum prolactin (mean +/- S.E.M.) in control ovariectomized rats was 4.0 +/- 0.9 micrograms/l and one and three injections of OB raised these levels to 14.4 +/- 5.0 and 28.8 +/- 4.6 micrograms/l respectively. The highest levels of serum prolactin were seen in sulpiride-treated rats (467.2 +/- 28.7 micrograms/l). Striking differences occurred in the pituitary contents of big prolactin, the control values increasing from 5.3 +/- 0.5 to 10.2 +/- 1.3 micrograms/mg after one OB injection and to 14.7 +/- 0.7 micrograms/mg after three OB injections.(ABSTRACT TRUNCATED AT 250 WORDS)

Regression of redundant lactotrophs in rat pituitary gland after cessation of lactation.

Regressive changes occurring in the pituitary gland of the rat after removal of litters were studied. Pituitary glands of lactating rats were characterized by the presence of numerous hypertrophied lactotrophs. Interruption of lactation caused a blockade of prolactin synthesis and secretion, followed by degeneration of lactotrophs. Morphometric analysis of pituitary glands revealed that lactotrophs accounted for about 50% of the total hypophysial cell count in lactating rats. This percentage decreased progressively and reached pre-pregnant levels 7 days after removal of litters; the decrease was inversely correlated with an increase in the number of degenerating lactotrophs which comprised 30% of all lactotrophs 72 h after removal of litters. The morphological changes found in lactotrophs were closely related to changes in the prolactin content of serum and the pituitary gland. Regression of lactotrophs appeared to be the most important cause inducing the reversal of hypophysial lactotrophic activity to pre-pregnant conditions.

Gene transfer into human leukemia cell lines by electroporation: experience with exponentially decaying and square wave pulse.

The efficiency of gene transfer into human leukemia cell lines by electroporation was investigated. For both transient expression (beta-galactosidase gene) and stable transformation (neomycin resistance gene), the transfer efficiency into leukemia cell lines using a square wave pulse was superior to that using an exponentially decaying wave. The transfer rate of pMoZtk (containing beta-galactosidase gene) into K562 by electroporation using a square wave was approximately 5%, compared with 1% by an exponentially decaying pulse. Whereas the transfer rate of pMAM-neo into K562 by electroporation using an exponentially decaying pulse was less than 10(-5), a square wave generated much more efficient introduction rate of nearly 10(-3). In the other leukemia cell lines also, some square wave yields were better than exponential yields and all square wave yields were at least as good as the exponential yields.

Gene introduction into granulocyte-macrophage progenitor cells by electroporation: the relationship between introduction efficiency and the proportion of cells in S-phase.

Our previous study demonstrated the positive relationship between the gene introduction rate into hematopoietic cell lines by electroporation and the percentage of cells in S-phase. In the present study, granulocyte-macrophage progenitor cells (CFU-C) rich marrow cell fraction were cultured in suspension with IL-3, GM-CSF and G-CSF for 4 days. The number of CFU-C were increased three times after the culture, and 3H-thymidine suicide tests of cultured cells demonstrated that the proportion of CFU-C in S-phase was increased by two to four times. The efficiency of gene transfer into CFU-C with the plasmid pMoZtk (containing the beta-galactosidase gene) by electroporation was nearly doubled by culturing marrow cells with these growth factors. These findings confirm that the introduction rate of the gene into CFU-C by electroporation is more efficient in cell populations with a higher percentage of CFU-C in S-phase.

Preferential proliferation of anti-DNA producing cells of NZB mice in NZB.xid recipients.

B cells from autoimmune NZB mice were transferred into unmanipulated non-autoimmune NZB.xid mice. The number of antibody-producing cells against various antigens in recipient mice was monitored at varying time after cell transfer using ELISPOT assay. NZB B cells producing antibody against all antigens we examined were able to proliferate in NZB.xid mice, which supports the idea of polyclonal B cell activation. However, anti-DNA producing cells proliferated most rapidly, and anti-BrMRBC producing cells proliferated more rapidly than B cells of other antigenic specificities. The percentage of anti-DNA producing cells in total immunoglobulin-producing cells increased over time whereas the percentage of anti-ovalbumin producing cells kept the same level. This indicates directly the preferential proliferation of NZB anti-DNA producing cells in NZB.xid mice. The result shows the responsibility of antigen-specific stimulation or activation on autoimmunity in the context of polyclonal B cell activation.