Selective synthesis of A-ring Е-arylidene derivatives from ß-sitosterol and their activity.

A series of 24-ethylcholest-4-ene-3,6-dione 2E-arylidene-derivatives has been synthesized by a Claisen-Schmidt reaction from a natural phytosterol ß-sitosterol with yields of 80-85%. The structure of the obtained compounds was confirmed by NMR spectroscopy, including two-dimensional correlation experiments. The synthesized compounds were evaluated for their in vitro cytotoxicity and α-glucosidase inhibitory activity. It was established that compound 3 with pyridin-3-ylmethylene moiety exhibited a selective cytotoxic effect against the U251 cancer cell line with 99.31% inhibition of cancer cell growth. Compounds with pyridin-4-ylmethylene 4 and furan-2-ylmethylene-5 fragments were the most active inhibitors of α-glucosidase with IC50 64.00 and 38.95 µM, being 3- and 5-times more active than acarbose. Binding mode to α-glucosidase and ADMET characteristics for the lead molecule 5 were proposed computationally. To sum up, an efficient approach to the derivatives with promising antidiabetic activity based on available natural product ß-sitosterol is suggested.

The Efficacy of Fetoscopic Laser Surgery in Twin-Twin Transfusion Syndrome: A Preliminary Vietnamese Study.

Background: Twin-twin transfusion syndrome (TTTS) is a severe prenatal complication of monochorionic diamniotic twins (MCDA). If left untreated, TTTS is associated with a high risk of neonatal death and neurological complications. Various treatment methods for TTTS have been proposed, but fetoscopic laser surgery (FLS) is currently the primary treatment for TTTS in global fetal medicine centers. The objective of this study was to evaluate the outcome of TTTS following FLS treatment at Hanoi Obstetrics and Gynecology Hospital (HOGH), a new fetal medicine center in Vietnam. Methods: A prospective study of a series of 33 consecutive TTTS cases prior to 26 weeks of gestation subjected to FLS at HOGH in Vie-tnam between September 2019 to July 2021. Neonates were monitored for at least six months after birth. Results: The survival rate of at least one fetus reached 84.85%. No short-term neurological complications have been reported upon follow-up of the newborn up to six months after birth. There were three stillbirth cases (9.09%), one case of preterm ruptured membra-nes (PROM) (3.03%) after seven days of surgery and three cases of recurrent TTTS after surgery using the Solomon technique (25%). No maternal complications were observed. The mean gestational age at birth was 33.76 ± 4.52 weeks, with a mean interval of 12.24 ± 6.67 weeks between intervention and delivery. Nine cases (30%) were born prematurely before 32 weeks. Additionally, 60% of recipients and nearly 90% of donors weighed less than 2500 grams. Conclusions: FLS leads to high survival rates for fetuses with TTTS. FLS seems to be an effective therapeutic option for TTTS before 26 weeks of gestation.

Obstetric and Perinatal Outcomes of Dichorionic-Diamniotic Twin Pregnancies Conceived by IVF/ICSI Compared with Those Conceived Spontaneously.

Introduction: This study aimed to investigate differences in mater-nal and perinatal outcomes between dichorionic-diamniotic (DCDA) twin pregnancies between those conceived spontaneously and those conceived through in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI). Methods: This study was a single-center, retrospective cohort study. All women with DCDA twin pregnancies were considered for inclusion. Monochorionic twins and higher-order multiple pre-gnancies were excluded. All data related to maternal and perinatal outcomes were extracted from the hospital database and compared between spontaneously conceived DCDA twin pregnancies and those conceived by IVF/ICSI. Multivariable logistic regression was used to adjust for confounders to determine factors associated with maternal and perinatal outcomes. Results: Of 739 identified DCDA twin pregnancies, 483 (65.4%) were conceived through IVF/ICSI treatment (IVF/ICSI group), and 256 (34.6%) were spontaneously conceived (SC group). Women in the IVF/ICSI group were older and had fewer previous live births than women in the SC group. The women in the IVF/ICSI group had significantly higher risks of preeclampsia (adjusted odds ratio [aOR]: 2.50; 95% confidence interval [CI]: 1.12-5.55), cesarean delivery (aOR: 2.0; 95% CI: 1.27-3.17), an postpartum hemorrhage following cesarean section (aOR: 3.15; 95% CI: 1.53-6.45). The DCDA twins in the IVF/ICSI group were delivered at an earlier gestational age (36.2 vs. 36.7 weeks, p < 0.001), had slightly lower mean birth weights (2298 vs. 2367 g, p = 0.005), and required more respiratory support (aOR: 0.69; 95% Cl: 0.48-0.98) than those in the SC group. Conclusions: Our study demonstrated that women with DCDA twin pregnancies conceived through IVF/ICSI experienced more complications than those with SC DCDA twin pregnancies. Newborns in the IVF/ICSI group had a slightly lower mean birth weight and required respiratory support more frequently, but no other significant differences in perinatal outcomes or perinatal mortality were observed between the two groups.

Efficacy of Rose Myrtle Rhodomyrtus tomentosa Seed Extract against Acute Hepatopancreatic Necrosis Disease in Pacific Whiteleg Shrimp Penaeus vannamei.

Acute hepatopancreatic necrosis disease (AHPND) is a new emerging bacterial disease that has been recently reported to cause mass mortalities in Pacific whiteleg shrimp Penaeus vannamei. Antibiotics have been used to treat bacterial diseases in shrimp, but most of them have been ineffective and have resulted in drug residues in the harvested shrimp products. In this study, an alternative approach was tested for its efficacy in controlling AHPND. The extract of rose myrtle Rhodomyrtus tomentosa seed, a traditional Vietnamese medicine, was tested for antibacterial effect against three AHPND bacterial strains in vitro (Vibrio parahaemolyticus [VPAHPND ] KC12.020, VPAHPND KC13.14.2, and V. harveyi KC13.17.5) and was further evaluated for its potential efficacy in prevention of AHPND in shrimp in vivo. The in vitro studies showed that the antibacterial activity of the R. tomentosa extract was dose dependent, with the strongest bacterial susceptibility (≥18.0 mm) at a concentration of around 3,500 µg/disc. The in vivo studies showed that after challenge with VPAHPND KC12.020, the survival rates for shrimp in the groups that received feed pellets supplemented with extract at 3.5% or 7.0% (survival ~48.9% and 52.2%, respectively) were significantly higher than the zero survival rate in the positive control group, which received feed without the extract. These results indicate that the use of the R. tomentosa extract as an alternative therapy for control of AHPND in shrimp could help to minimize disease outbreaks. As a result, the extract is further expected to reduce drug/chemical residues in shrimp products.

Analysis of porcine body size variation using re-sequencing data of miniature and large pigs.

BACKGROUND: Domestication has led to substantial phenotypic and genetic variation in domestic animals. In pigs, the size of so called minipigs differs by one order of magnitude compared to breeds of large body size. We used biallelic SNPs identified from re-sequencing data to compare various publicly available wild and domestic populations against two minipig breeds to gain better understanding of the genetic background of the extensive body size variation. We combined two complementary measures, expected heterozygosity and the composite likelihood ratio test implemented in "SweepFinder", to identify signatures of selection in Minipigs. We intersected these sweep regions with a measure of differentiation, namely FST, to remove regions of low variation across pigs. An extraordinary large sweep between 52 and 61 Mb on chromosome X was separately analyzed based on SNP-array data of F2 individuals from a cross of Goettingen Minipigs and large pigs. RESULTS: Selective sweep analysis identified putative sweep regions for growth and subsequent gene annotation provided a comprehensive set of putative candidate genes. A long swept haplotype on chromosome X, descending from the Goettingen Minipig founders was associated with a reduction of adult body length by 3% in F2 cross-breds. CONCLUSION: The resulting set of genes in putative sweep regions implies that the genetic background of body size variation in pigs is polygenic rather than mono- or oligogenic. Identified genes suggest alterations in metabolic functions and a possible insulin resistance to contribute to miniaturization. A size QTL located within the sweep on chromosome X, with an estimated effect of 3% on body length, is comparable to the largest known in pigs or other species. The androgen receptor AR, previously known to influence pig performance and carcass traits, is the most obvious potential candidate gene within this region.

Liver transcriptome analysis reveals important factors involved in the metabolic adaptation of the transition cow.

During early lactation, dairy cows experience a severe metabolic load often resulting in the development of various diseases. The inevitable deficiency in nutrients and energy at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. We conducted a whole-liver transcriptome analysis for the transition cow to identify novel factors crucial for metabolic adaptation. Liver samples were obtained from 6 Red Holstein dairy cows (parity 2 to 7, mean ± standard deviation: 3.7 ± 2.3) at 3 time points: T1 = 22 ± 4 d antepartum, T2 = 10 ± 2 d postpartum, and T3 = 17 ± 2 d postpartum. Using RNA sequencing (RNA-seq), we studied the transcriptomic profile of the transition cow before and after parturition. We performed a differential gene expression analysis (DGEA) and gene-set enrichment analysis (GSEA) for biological processes (gene ontology, GO) and pathways (Kyoto Encyclopedia of Genes and Genomes, KEGG). Among the 10,186 expressed genes, we discovered 1,063 differentially expressed genes (false discovery rate = 5%). The GSEA revealed 16 biological processes and 7 pathways significantly (false discovery rate = 5%) associated with the hepatic changes of the transition cow. Our results confirm that major hepatic changes are related to energy mobilization after parturition; in particular, they are related to fatty acid oxidation/metabolism, cholesterol metabolism, and gluconeogenesis. Using the STRING database (https://string-db.org/), we investigated interactions between significant genes and identified 9 key genes (CYP7A1, APOA1, CREM, LOC522146, CYP2C87, HMGCR, FDFT1, SGLE, and CYP26A1) through which the different processes involved in the metabolic adaptation interact. Comparing our main results with the literature, we could identify further genes that have not yet been associated with the transition period (e.g., CPT1B, ADIPOR2, LEPR, CREB3L3, and CCND1) and that are mainly involved in processes controlled by AMP-activated protein kinase, an important regulator of energy homeostasis.

A reaction norm sire model to study the effect of metabolic challenge in early lactation on the functional longevity of dairy cows.

Due to the discrepancy of the high energy demand for rapidly increasing milk production and limited feed intake in the transition period around parturition, dairy cows require considerable metabolic adaptations. We hypothesize that some cows are genetically less suited to cope with these metabolic needs than others, leading to adverse follow-up effects on longevity. To test this, we designed a reaction norm model in which functional lifetime was linked to the metabolic challenge in the beginning of the first lactation. As challenge variables, we used either the sum of milk yield or the accumulated fat-to-protein ratio of the first 3 test-days (<120 d in milk), pre-adjusted for herd-test-day variance. We defined a random regression sire model, in which a random slope was estimated for each sire to assess whether a bull had robust (neutral or positive slopes) or non-robust (negative slopes) daughters. We fitted the model to data of ∼580,000 daughters of ∼5,000 Brown Swiss bulls with suitable observations available (≥10 daughters per bull). To validate our proposed model and assess the reliability of the estimated (co)variance components, we conducted an extensive bootstrap approach. For both challenge variables, we found the sire variance for the slope of the random regression to be significantly different from zero, suggesting a genetic component for metabolic adaptability. The results of the study show that the ability to cope with metabolic stress in the transition period has a genetic component, which can be used to breed metabolically robust dairy cows.

Genetic structure of the threatened Dipterocarpus costatus populations in lowland tropical rainforests of southern Vietnam.

Dipterocarpus costatus is an endangered species restricted to the lowland forests of southern Vietnam. Habitat loss and over-exploitation of D. costatus wood are the major threats to this species. We investigated the level of genetic variability within and among populations of D. costatus in order to provide guidelines for the conservation, management, and restoration of this species to the Forest Protection Department, Vietnam. Nine microsatellite markers were used to analyze 114 samples from four populations representing the natural range of D. costatus in southeast Vietnam. We indicated the low allelic diversity (NA = 2.3) and low genetic diversities with an average observed and expected heterozygosity of 0.130 and 0.151, respectively, in the lowland forests of southeast Vietnam. The low genetic diversity might be a consequence of inbreeding within the small and isolated populations of D. costatus owing to its habitat loss and over-exploitation. All populations deviated from Hardy-Weinberg equilibrium showing reduced heterozygosity. Alleles were lost from the populations by genetic drift. Genetic differentiation among populations was high (average pairwise FST = 0.405), indicating low gene flow (<1) and isolated populations due to its destructed habitat and large geographical distances (P < 0.05) among populations. Heterozygosity excess tests (except of Bu Gia Map only under infinite allele model) were negative. The high genetic variation (62.7%) was found within populations. The STRUCTURE and neighbor joining tree results suggest strong differentiation among D. costatus populations, with the three genetic clusters, Phu Quoc, Tan Phu and Bu Gia Map, and Lo Go-Xa Mat due to habitat fragmentation and isolation. The threatened status of D. costatus was related to a lack of genetic diversity, with all its populations isolated in small forest patches. We recommend the establishment of an ex situ conservation site for D. costatus with a new big population comprising all genetic groups in order to enhance its survival under different environmental stresses.

Spatial heterogeneous distribution of SiO(x) → SiO(x±1) reactions in silica liquid.

We have numerically studied the diffusion mechanism in silica liquid following an approach where the diffusion rate is evaluated via the SiO(x) → SiO(x±1) reaction rate υ(react) and the mean square displacement of particles d(react) as a reaction happens. Five models at pressure up to 25 GPa and at a temperature of 3000 K have been constructed by molecular dynamic simulation. When applying pressure to the liquid, υ(react) increases monotonously because the Si-O bond becomes weaker with pressure. Meanwhile d(react) attains a maximum near the point of 10 GPa despite the particles move in a significantly smaller volume. Furthermore, the SiO(x) → SiO(x±1) reactions are spatially heterogeneously distributed in the liquid. Upon low pressure, most reactions happen with a small number of Si particles. This reaction localization causes the diffusion anomaly and dynamics heterogeneity in the liquid. With increasing pressure the diffusion mechanism changes from the heterogeneous spatial distribution of reactions to homogeneous one. The simulation also reveals two distinguished regions with quite different coordination environments where the reaction rate significantly differs from each other. These sets of Si particles migrate in space over time and form regions with so-called "fast" and "slow" Si particles. The result obtained here indicates the coexistence of low- and high-density regions, and supports the concept of polymorphism in silica liquid.

Computer simulation of diffusion in silica liquid under temperature and pressure.

We have studied the diffusion mechanism in silica liquid following a new approach where the diffusion rate is estimated via the rate of SiO(x) → SiO(x±1) and the mean square displacement of Si particles per SiO(x) → SiO(x±1). Molecular dynamics simulation has been conducted for a model consisting of 1998 particles over a wide range of temperatures (3000-4500 K) and pressure (from 0 to 25.75 GPa). Our results show that the rate of SiO(x) → SiO(x±1) increases either with increasing the temperature or pressure. Further, we find that SiO(x) → SiO(x±1) is heterogeneously distributed through the network structure of the liquid. In particular, it is concentrated on a small section of Si particles in a low-temperature regime and at ambient pressure. The spatial localisation of SiO(x) → SiO(x±1) originates from the fact that the stable unit in low- and high-pressure regime is SiO4 and SiO6, respectively. The major change in the diffusion mechanism under pressure or temperature concerns the change in the distribution of SiO(x) → SiO(x±1) through the network structure. It is finally shown that the spatial localisation of SiO(x) → SiO(x±1) is responsible for the dynamics heterogeneity and the diffusion anomaly for silica liquid. This finding supports the concept that as the temperature approaches the glass transition point, SiO(x) → SiO(x±1) spatially localises such that the diffusivity drops and the dynamics are anomalously slow.

Correlation effect for dynamics in silica liquid.

We study numerically the diffusion mechanism in silica liquid via molecular dynamics simulation. For this purpose we examine the evolution of structural units SiO(x) (x=4-6) for different times and at temperatures from 3000 to 4500 K. Simulation shows that the diffusivity of the silicon particle is performed through the transition SiO(x)→SiO(x±1), i.e., the bond-breaking and bond-reformation events. As a SiO(x)→SiO(x±1) transition occurs, one oxygen particle moves out of or into the coordination shell, leading to a collective movement of Si particles. Other types of transitions, for instance, SiO(4)→SiO(6) or SiO(6)→SiO(4), are negligible. We establish an expression for the diffusion coefficient that shows that the diffusivity is not proportional to the rate of SiO(x)→SiO(x±1) because it is strongly localized in the network structure. A high degree of localization of SiO(x)→SiO(x±1) leads to a heterogeneous dynamics. We find that the dynamics slowdown is determined by two terms: The first one concerns the change in the statistic property related to the fraction of non-four-coordinated units (SiO(3), SiO(5), SiO(6), and SiO(7)) and the second term concerns the correlation effect. Furthermore, we show that the correlation coefficient depends on both the fraction of the back-forth SiO(x)→SiO(x±1) transition and the degree of localization of SiO(x)→SiO(x±1). Our finding qualitatively supports the ideal that anomalously slow dynamics near the glass-transition point is caused by a strong localization of SiO(x)→SiO(x±1).

Inhibition by Artocarpus tonkinensis of the development of collagen-induced arthritis in rats.

Extracts of the leaves and roots from the tree Artocarpus tonkinensis A Cheval (family Moraceae) are used in traditional Vietnamese medicine in order to treat backache as well as rheumatic joint diseases. We prepared an ethyl acetate (EtOAc) extract from this plant and tested its anti-inflammatory properties in an experimental arthritis model, collagen-induced arthritis (CIA). CIA was induced in Dark Agouti rats by means of immunization with collagen type II (CII) emulsified in incomplete Freund's adjuvant. Starting at the day of immunization, the rats were treated daily with intraperitoneal injections of Artocarpus extract. Arthritis progression was measured by means of clinical scoring of paws and anti-CII antibody titres were measured by means of ELISA. In vitro, lymph node (LN) cell cultures were treated with Artocarpus extract and the apoptosis-inducing effect was determined with FACS staining by using annexin V and propidium iodide as well as the TUNEL method. Treatment of the rats with Artocarpus extract decreased arthritis incidence and severity and delayed disease onset. When treatment was started after the onset of arthritis, a tendency towards arthritis amelioration was observed. In vitro, Artocarpus extract acted as a T-cell modulator, inhibiting mitogen-induced T-cell proliferation and inducing apoptosis of activated LN-derived lymphocytes. Thus, we have demonstrated that an EtOAc extract of Artocarpus, a plant traditionally used in Vietnamese folk medicine for treating arthritic conditions, has beneficial effects in an experimental arthritis model. This effect is likely to be T cell-dependent and mediated through apoptosis induction in activated cells.

239+240Pu, 90Sr and 137Cs inventories in surface soils of Vietnam.

Fallout 239+240Pu, 238Pu, 90Sr and 137Cs inventories in surface soils were measured for 20 locations in northern Vietnam yielding the mean values (+/- standard error) of 26.5+/-3.8 Bq m(-2) for 239+240Pu, 1048+/-143 Bq m(-2) for 137Cs and 212+/-28 Bq m(-2) for 90Sr. The concentrations of 137Cs and plutonium isotopes strongly correlate with each other resulting in a stable 239+240Pu/137Cs inventory ratio of 0.025+/-0.002. Among soil parameters, organic matter and fulvic acids strongly correlate with caesium and plutonium isotopes, especially in the 0-10 cm layer. 137Cs and 239+240Pu are distributed rather similarly over the 0-10 cm and 10-20 cm layers. At locations with high contents of sand (82-93%) along the South China Sea coast, the downward percolation by rainwater results in a higher accumulation of 239+240Pu and 137Cs in the 10-20 cm layer. The mean 137Cs/ 90Sr inventory ratio is 9.3+/-2.2, and the correlation is weak between these isotopes.

H626R and R124C mutations of the TGFBI (BIGH3) gene caused lattice corneal dystrophy in Vietnamese people.

BACKGROUND/AIMS: Mutations of the human transforming growth factor beta induced gene (TGFBI) were reported to cause lattice corneal dystrophy (LCD) in various nationalities. This study analysed the TGFBI gene in Vietnamese people with LCD. METHODS: 13 unrelated families, including 34 patients and 21 unaffected members were examined. 50 normal Vietnamese people served as controls. Blood samples were collected. Genomic DNA was extracted from leucocytes. Analysis of TGFBI gene was performed using the polymerase chain reaction and direct sequencing. Corneal buttons were studied histopathologically. RESULTS: Two clinically distinguishable forms of LCD were revealed: one was typical of LCDI; the other was characterised by the late onset, thick lattice lines, and asymmetry between two eyes. Sequencing of the TGFBI gene revealed R124C mutation in three families and H626R mutation in 10 families. Congo red staining of the H626R-LCD cornea showed amyloid deposits in the subepithelial and stromal layers. CONCLUSIONS: R124C and H626R mutations of TGFBI gene caused LCD in Vietnamese people. R124C, a common cause of LCDI in many nationalities, was relatively rare, whereas H626R reported in several white people but not yet in Asians was most common (>75%) in Vietnamese people. Since the phenotype caused by H626R represents a new variant intermediate between LCDI and LCDIIIA, we proposed to consider it as LCD type IIIB.

Molecular epidemiology of pneumococcal carriage among children with upper respiratory tract infections in Hanoi, Vietnam.

To investigate the molecular epidemiology of pneumococcal nasopharyngeal carriage in Hanoi, Vietnam, we studied 84 pneumococcal strains retrieved from children with upper respiratory tract infections. Serotypes 23F (32%), 19F (21%), 6B (13%), and 14 (10%) were found most often. A significant number of strains were antibiotic resistant. Fifty-two percent of the strains were (intermediate) resistant to penicillin, 87% were (intermediate) resistant to co-trimoxazole, 76% were resistant to tetracycline, 73% were resistant to erythromycin, and 39% were (intermediate) resistant to cefotaxime. Seventy-five percent were resistant to three or more classes of antibiotics. A high degree of genetic heterogeneity among the penicillin resistance genes was observed. In addition, the tetracycline resistance gene tet(M) and the erythromycin resistance gene erm(B) were predominantly observed among the isolates. Molecular analysis of the 84 isolates by restriction fragment end labeling (RFEL) revealed 35 distinct genotypes. Twelve of these genotypes represented a total of eight genetic clusters with 61 isolates (73%). The two largest clusters contained 24 and 12 isolates, and the isolates in those clusters were identical to the two internationally spreading multidrug-resistant clones Spain 23F-1 and Taiwan 19F-14, respectively. The remaining RFEL types were Vietnam specific, as they did not match the types in our reference collection of 193 distinct RFEL types from 16 countries. Furthermore, 57 of the 61 horizontally spreading isolates (93%) in the eight genetic clusters were covered by the seven-valent conjugate vaccine, whereas this vaccine covered only 43% of the isolates with unique genotypes. According to the serotype distribution of the nasopharyngeal pneumococcal isolates, this study suggests a high potential benefit of the seven-valent pneumococcal conjugate vaccine for children in Hanoi.

[Expression of estrogen receptors alpha and beta, androgen receptors and progesterone receptors in human cornea].

PURPOSE: Using immunohistochemical techniques and reverse transcription polymerase chain reaction (RT-PCR), we examined the localization of estrogen receptors alpha and beta (ER alpha, beta), androgen receptor (AR), and progesterone receptor (PR) in human corneas. MATERIALS AND METHODS: Using formalin-fixed donor human cornea, we did immunohistochemical staining after making transverse sections, and examined the localization of receptors. Also, we extracted mRNA from primary culture cells of the corneal epithelium and stroma as well as the endothelial cell layer and epithelial layer of the cornea, and we performed RT-PCR and examined the expression of each receptor. RESULTS: Immunohistochemical staining revealed that ER alpha was localized in corneal epithelial cells as well as in corneal stromal cells, and ER beta, AR and PR were localized in corneal epithelial, stromal, and endothelial cells. ER alpha, ER beta, and AR mRNA expression was observed in cultured and in vivo epithelium and cultured stroma cells. PR mRNA was expressed not only in cultured and in vivo epithelium and in cultured stroma cells but also in endothelium. CONCLUSIONS: We detected the localization of estrogen receptors alpha and beta, androgen receptors, and progesterone receptors in the human cornea.

Characterization of trichobakin, a type I ribosome-inactivating protein from Trichosanthes sp. Bac Kan 8-98.

We have isolated a genomic clone encoding trichobakin (TBK), a type I ribosome-inactivating protein from the plant Trichosanthes sp. Bac Kan 8-98 (family Cucurbitaceae), by PCR using specific primers designed from the cDNA sequences of alpha-trichosanthin. The sequence encoding mature TBK was constructed in the pET-21d(+) vector for overexpression in Escherichia coli strain BL21(DE3). The recombinant protein was purified to homogeneity by CM-Sepharose chromatography on FPLC with a final yield of about 55 mg/l of culture. The protein has a molecular mass of about 27 kDa, as shown by SDS/PAGE and matrix-assisted laser-desorption ionization MS. It was found that the protein inhibited luciferase mRNA translation in the rabbit reticulocyte cell-free system with an IC(50) value (that which causes a 50% reduction of residual translational activity) of about 3.5 pM. The rRNA N-glycosidase activity of the protein was also proved at the above-mentioned concentration after rRNAs were treated with acid aniline.

Six different mutations of TGFBI (betaig-h3, keratoepithelin) gene found in Japanese corneal dystrophies.

PURPOSE: To investigate mutations of the human transforming growth factor beta-induced gene (TGFBI), transforming growth factor-beta-induced gene product (betaig-h3, keratoepithelin), in Japanese patients with Avellino corneal dystrophy (ACD), lattice corneal dystrophy (LCD), granular corneal dystrophy (GCD), and Reis-Bücklers corneal dystrophy (RBCD). METHODS: Genomic DNA was extracted from the peripheral blood of 75 patients and 7 unaffected relatives from 60 families with ACD, 34 patients and 8 unaffected relatives from 21 families with LCD, 4 patients and 4 unaffected relatives from 4 families with GCD, and 4 patients and an unaffected relative from 3 families with RBCD. Fifty normal volunteers served as controls. Exons 4, 11, and 12 of the TGFBI gene were amplified by polymerase chain reaction and were directly sequenced. RESULTS: Six different heterozygous missense mutations were detected in codons R124, L518, L527, and R555 of the TGFBI gene in the 117 patients from 88 families. A R124H mutation was detected in the patients with ACD. A R124C mutation was detected in the patients with LCD type 1 (LCD1), L518P was in atypical LCDI, and L527R in LCD with opacities deep in stroma. A R555W mutation was detected in the patients with GCD. A R555Q mutation was detected in the patients with RBCD. CONCLUSIONS: We conclude that codons R124 and R555 of the TGFBI gene are also hot spots in Japanese patients with ACD, LCD, GCD, and RBCD. Many Japanese patients with CD had ACD with R124H mutation. GCD with R555W mutation was rare.

Q118X mutation of M1S1 gene caused gelatinous drop-like corneal dystrophy: the P501T of BIGH3 gene found in a family with gelatinous drop-like corneal dystrophy.

PURPOSE: To analyze BIGH3 and M1S1 genes in two Japanese brothers with gelatinous drop-like corneal dystrophy and five unaffected family members. METHODS: DNA was extracted, and each part of the two genes was amplified and directly sequenced. RESULTS: On the BIGH3 gene, a heterozygous P501T mutation was found in the elder brother and three unaffected family members. On the M1S1 gene, both brothers with gelatinous drop-like corneal dystrophy showed a homozygous Q118X mutation, whereas all unaffected members were heterozygous. CONCLUSIONS: The Q118X mutation of M1S1 gene caused gelatinous drop-like corneal dystrophy. Although the P501T of the BIGH3 gene found in this pedigree was precisely the one reported for lattice corneal dystrophy IIIA, no clinical feature was shown, even in the 85-year-old father. This fact shows that the P501T mutation for LCDIIIA has low penetrance.

Fluoride profiles in dental plaque in vivo formed on fluoride pre-treated human enamel.

Using a novel device capable of generating plaque in vivo on a natural enamel substrate, it has been possible to determine fluoride profiles from the saliva-plaque interface towards the enamel surface. Fluoride profiles in dental plaques tended to fall from the saliva-plaque interface towards the enamel. The device also offered the possibility of examining fluoride distributions after pre-treatment of the enamel with fluoride in vitro. Fluoride profiles were determined in plaque generated in vivo on enamel surfaces, which had been previously treated with a 900-ppm fluoride solution. The results showed the previously reported fall from the plaque surface, but in addition, a further rise towards the enamel surface was seen. The data imply that enamel loaded with fluoride can release some of this fluoride back into the plaque and may act as a fluoride reservoir.