A new paradigm emerges from the study of de novo mutations in the context of neurodevelopmental disease.

The study of de novo point mutations (new germline mutations arising from the gametes of the parents) remained largely static until the arrival of next-generation sequencing technologies, which made both whole-exome sequencing (WES) and whole-genome sequencing (WGS) feasible in practical terms. Single nucleotide polymorphism genotyping arrays have been used to identify de novo copy-number variants in a number of common neurodevelopmental conditions such as schizophrenia and autism. By contrast, as point mutations and microlesions occurring de novo are refractory to analysis by these microarray-based methods, little was known about either their frequency or impact upon neurodevelopmental disease, until the advent of WES. De novo point mutations have recently been implicated in schizophrenia, autism and mental retardation through the WES of case-parent trios. Taken together, these findings strengthen the hypothesis that the occurrence of de novo mutations could account for the high prevalence of such diseases that are associated with a marked reduction in fecundity. De novo point mutations are also known to be responsible for many sporadic cases of rare dominant mendelian disorders such as Kabuki syndrome, Schinzel-Giedion syndrome and Bohring-Opitz syndrome. These disorders share a common feature in that they are all characterized by intellectual disability. In summary, recent WES studies of neurodevelopmental and neuropsychiatric disease have provided new insights into the role of de novo mutations in these disorders. Our knowledge of de novo mutations is likely to be further accelerated by WGS. However, the collection of case-parent trios will be a prerequisite for such studies. This review aims to discuss recent developments in the study of de novo mutations made possible by technological advances in DNA sequencing.

Integrating next-generation sequencing into the diagnostic testing of inherited cancer predisposition.

The clinical application of next-generation sequencing (NGS) as a diagnostic tool has become increasingly evident. The coupling of NGS technologies with new genomic sequence enrichment methods has made the sequencing of panels of target genes technically feasible, at the same time as making such an approach cost-effective for diagnostic applications. In this article, we discuss recent studies that have applied NGS in the diagnostic setting in relation to hereditary cancer.

Antibacterial activity of hydroxyalkenyl salicylic acids from sarcotesta of Ginkgo biloba against vancomycin-resistant Enterococcus.

A chloroform fraction prepared from the sarcotesta of Ginkgo biloba showed potent inhibitory activity against vancomycin-resistant Enterococcus (VRE). The active compounds were elucidated to be 2-hydroxy-6-(8-pentadecenyl) salicylic acid (1) and 2-hydroxy-6-(10-heptadecenyl) salicylic acid (2) based on their spectral analysis. Compounds 1 and 2 showed significant antibacterial activities against VRE.

Optimization of the extraction of anthocyanin from Bokbunja (Rubus coreanus Miq.) marc produced during traditional wine processing and characterization of the extracts.

The extraction of anthocyanin from Bokbunja (Rubus coreanus Miq.) marc generated during traditional wine processing was optimized using response surface methodology (RSM). A face-centered cube design (FCD) consisting of 17 experimental runs, including five replicates at the center point, was used to investigate the effects of the three variables (solid-liquid ratio, time, and temperature) on anthocyanin extraction, and the results showed that the relationship between the three variables and the total anthocyanin content followed a quadratic model (R2=0.8853). In addition, the RSM analysis predicted that the optimum conditions for extraction consisted of a solid-liquid ratio of 20, a time of 60min, and a temperature of 60 degrees C. Verification tests performed under these optimum conditions gave 34.7+/-1.4mg/100g of anthocyanin, which was close to predicted value of 37.2mg/100g. Additionally, analysis of water extracts prepared using the predicted optimum conditions revealed that the carbohydrates (sugar and pectin) in Bokbunja marc underwent significant variation toward the formation of by-products (glycerol and uronic acids) during yeast fermentation, and that the amount of anthocyanin produced was reduced 10-fold when compared to the original extraction. Further, the results of HPLC-PDA-MS/MS analysis of the anthocyanins extracted from Bokbunja marc revealed the presence of six anthocyanin components, which were tentatively identified as cyanidin 3-O-sambubioside, cyanidin 3-O-xylosylrutinoside, cyanidin 3-O-rutinoside, pelargonidin 3-O-rutinoside, delphinidin 3-O-rutinoside-?, and delphinidin 3-O-glucuronide.

Characterization of seed oils from fresh Bokbunja (Rubus coreanus Miq.) and wine processing waste.

The physicochemical characteristics, fatty acid (FA) profile, and triacylglyceride (TAG) composition of seed oils from fresh Bokbunja (Rubus coreanus Miq.) fruits and traditional Bokbunja wine processing waste were determined in this study. Oil contents of the fresh seeds and the seeds from wine processing waste were similar, accounting for about 18% of dry weight. The free fatty acid (FFA) content between the two seed oils was significantly different (0.50% for fresh seed oil and 73.14% for wine seed oil). Iodine, conjugated diene, saponification values, and unsaponifiable matter were very similar in the oil samples, but the specific extinction coefficients at 232 and 270 nm of wine seed oil were higher than those of fresh seed oil. Linoleic (C18:2, 50.45-53.18%, L) and linolenic (C18:3, 29.36-33.25%, Ln) acids were the dominant FAs in the two seed oils, whereas oleic (C18:1, 7.32-8.04%, O), palmitic (C16:0, 1.55-1.65%, P), and stearic (C18:0, 0.65-0.68%, S) acids were the minor FAs. LLL, OLL, LLLn, OOL, LLnLn, and OOO were the abundant TAGs, representing >90% of the oils.

Binding affinity of proanthocyanidin from waste Pinus radiata bark onto proline-rich bovine achilles tendon collagen type I.

Binding affinity of proanthocyanidin (PA) purified from Pinus radiata bark waste onto bovine Achilles tendon collagen (type I) was studied. Adsorption of PA onto the collagen was optimized by examining pH, contact time and temperature. The adsorption was pH-dependent. The maximum adsorption capacity (Q(0)) of PA on collagen was found to be 211 mg g(-1) using the Langmuir isotherm. Comparison between two adsorbents also showed that collagen had higher adsorptivity of approximately 20% more than PVPP (polyvinyl polypyrrolidone). The high affinity between PA and collagen was further confirmed in solvent solubility experiments. The observed solvent resistance was thought to be mainly due to a hydrophobic stacking mechanism reinforced by hydrogen bonding. FT-IR spectra clearly indicated the presence of PA adsorbed on collagen. The results have interesting implications that PA can be a good protective agent for collagen against collagenase and other enzymes.

Optical properties and carrier dynamics of self-assembled GaN/Al(0.11)Ga(0.89)N quantum dots.

GaN quantum dots were grown on an Al(0.11)Ga(0.89)N buffer layer by using flow rate modulation epitaxy. The Stranski-Krastanov growth mode was identified by an atomic force microscopy study. The thickness of the wetting layer is about 7.2 monolayers. The temperature dependent photoluminescence studies showed that at low temperature the localization energy, which accounts for de-trapping of excitons, decreases with the reducing dot size. The decrease in emission efficiency at high temperature is attributed to the activation of carriers from the GaN dot to the nitrogen vacancy (V(N)) state of the Al(0.11)Ga(0.89)N barrier layer. The activation energy decreases with reducing dot size.

Anti-HBc IgG subclasses in different populations by comparing a variety of ELISA plates.

The number of IgG subclasses for hepatitis B virus (HBV) core antigen (anti-HBc), demonstrated for HBV-infected individuals, was measured by enzyme-linked immunosorbent assay (ELISA). Four commercially available hepatitis B core antigen (HBcAg) plates and one prepared plate were tested for ELISA sensitivity by the detection of 14 serum samples drawn from HBV chronic carriers, cured patients, vaccinees, and non-infected individuals. Differences in optical density (OD) values were obtained by comparing data gathered from the five plate types, suggesting that different plates may have different binding capabilities for each anti-HBc IgG subclass and, thus, contribute to the different ELISA sensitivities. Of these plates, the GB plate showed the most obvious absorbance changes for anti-HBc subclasses in different populations. These data also indicated different patterns for IgG-specific subclasses for various populations. For HBsAg+ carriers, the OD for IgG1 was greater than for IgG3. By contrast, the OD for IgG3 was higher than that for IgG1 in those subjects who were negative for HBsAg.

Circadian and weekly variations in pain onset of acute myocardial infarction.

BACKGROUND: A seasonal variation with more myocardial infarctions in the winter months due to cold weather has been reported. Other reports have described excess numbers of acute myocardial infarction (AMI) in the summer in Southern USA due to hot temperatures. To determine whether circadian and seasonal variations affect the incidence of AMI in the subtropical area of Taiwan, 480 consecutive patients with AMI admitted to our coronary care units were analyzed. METHODS: Six-hourly intervals over 24 hours (4 periods), daily intervals in a week (7 days) and monthly intervals in a year (12 months) were respectively studied. The distributions of the numbers of AMI occurring in the six-hour intervals were tested for differences among the four periods using the chi-squared test for goodness of fit. RESULTS: We found that there was a circadian variation in the onset of AMI with a morning peak (6 am to noon) (35%, chi 2 = 28.52, df = 3, p < 0.01) but no secondary late evening peak. The incidence of AMI was significantly lower on Sundays (9%) than on the other weekdays (chi 2 = 16.37, df = 6, p = 0.012). However, no seasonal variation (no winter or summer peaks) occurred in the incidence of AMI in this study (chi 2 = 0.77, df = 3, p = 0.99). CONCLUSION: Our results showed that there was a predominant morning peak in the onset of AMI. The low incidence of AMI cases on Sunday compared with other weekdays suggested that relief from tension or workload on Sundays might have an important role in this low percentage of AMI. Differing from other reports, there was no seasonal variation in the occurrence of AMI in our study, suggesting that the warm climate of a subtropical region does not provide an environment that is likely to increase the frequency of AMI.

Meningoencephalitis, myocarditis and disseminated intravascular coagulation in a patient with scrub typhus.

A 21-year-old male soldier was admitted due to a sore throat, headache, generalized lymphadenopathy and persistent fever for 12 days. Despite empirical antibiotic treatment for four days at a clinic prior to admission, he continued to have persistent abdominal pain over his right upper quadrant region and progressive jaundice was followed by shock. After admission, he developed an episode of clonic seizures and became delirious and agitated. An electrocardiogram showed first degree atrioventricular (AV) block and non-specific ST-T wave changes. Hematological studies revealed thrombocytopenia, hypofibrinogenemia, abnormal partial thromboplastin time (PTT) and a positive test for D-dimer. The cerebrospinal fluid analysis showed pleocytosis with white cells of 84/mm3 with a lymphocyte predominance, protein of 97 mg/dL and glucose of 79 mg/dL. Indirect immunofluorescence assay showed a fourfold rise in antibodies to Orientia tsutsugamushi in paired serum with IgM antibody titer of 1:640. The patient had a favorable response after parenteral chloramphenicol in addition to oral tetracycline. Early ricognition of scrub typhus and early prescription of anti-rickettsial agents prevent complications of central nervous system involvement and further deterioration of cardiac and hematological function.

Absence of a seasonal variation in myocardial infarction onset in a region without temperature extremes.

To evaluate whether the incidence of acute myocardial infarction (AMI) attack would have circadian, weekly and monthly variations in a subtropical area, 540 consecutive patients with AMI who were admitted to our coronary care units were analyzed. Six-hour intervals over 24 h, daily intervals in a week (7 days) and monthly intervals in a year (12 months) were studied, respectively. Results showed that there was a circadian variation in the onset of AMI with a morning (6 a.m. to noon) peak (34%, p < 0.01) but no secondary late evening (18-24 p.m.) peak. The incidence of AMI was significantly lower on Sundays (9%) than on the other weekdays (Monday through Saturday; p < 0.05). However, no monthly and seasonal variations in the incidence of AMI (no winter or summer peaks) were observed in this series. This study demonstrated a circadian variation in the onset of AMI attack with a predominant morning peak. The fewer AMI cases on Sundays compared to the other weekdays suggested that freedom from 'stress' or 'work-load' on Sundays might have an important impact on this low incidence of AMI. Unlike the large ranges in cold or hot weather found in temperate regions, the warm climate of a subtropical region does not affect the frequency of AMI.

Transesophageal color Doppler flow imaging in the evaluation of prosthetic cardiac valves.

To determine the value of transesophageal echocardiography in the assessment of prosthetic cardiac valves, 11 patients with clinically suspected cardiac prosthetic valve dysfunction were studied by transesophageal two-dimensional imaging, as well as by color Doppler flow mapping. Among these 11 patients, there were 10 with biological tissue valves and 3 with metallic valves (1 Bjork-Shiley, 2 St. Jude). Nine patients had replacement of mitral valves alone. The remaining two had received both mitral and aortic prostheses. The degree of mitral regurgitation was graded by transesophageal color Doppler according to the area of the regurgitant jet visualized. The degree of aortic regurgitation was graded by the jet height/left ventricular outflow height ratio method. All transesophageal studies were performed without complication and all were well tolerated. The pathological morphology of the cardiac prosthesis was clearly visualized by transesophageal two-dimensional imaging and subsequently proven at surgery. Of those tested, one patient was found to have a torn leaflet, one had a dislodged leaflet, one patient had paravalvular leakage, four had cusp vegetations, and five patients had prosthetic degeneration for other reasons. Mitral regurgitation was graded as absent in one patient, mild in two patients, moderate in two patients, and severe in six patients. Aortic regurgitation was graded as severe in both patients with aortic prostheses. We conclude that in patients with clinically suspected cardiac prosthetic dysfunction, transesophageal two-dimensional imaging combined with color Doppler can provide reliable information that corresponds to surgical findings.

Spontaneous contrast in the pericardial sac caused by gas-forming organisms.

Purulent pericarditis is a life-threatening disease associated with a variety of microorganisms. The case presented herein has unusual manifestations of contrast echoes on two-dimensional echocardiography. To our knowledge, this is the first reported case of pericarditis caused by gas-forming group D Streptococcus and Klebsiella pneumoniae with pyopericardium, resulting in contrast echoes on the two-dimensional echocardiographic image.

Detection of dysfunctioning mitral prosthetic regurgitation: transesophageal echocardiography in comparison with transthoracic echocardiography and angiography.

Transesophageal echocardiography has been increasingly used in addition to the transthoracic approach. It not only allows an unobstructed window for viewing the left atrium, but also provides high-quality imaging to improve diagnosis of prosthetic valve dysfunction. To determine the value of transesophageal echocardiography in detection of dysfunctioning mitral prosthetic regurgitation, 11 patients clinically suspected of dysfunctioning mitral prosthesis were studies by transthoracic, transesophageal echocardiography and was documented by a left ventricular angiography. There were 7 biological tissue valves and 4 metallic Bjork-Shiley valves. Compared with a left ventricular angiography, the transthoracic approach to 8 patients made an underestimation of the grade of regurgitation. The angiographic grading confirmed the transesophageal approach in all patients except one who was graded as mild regurgitation by left ventricular angiography, moderate regurgitation by transesophageal echocardiography, but no regurgitation by transthoracic echocardiography. Our results indicated that in patients with suspected mitral prosthetic dysfunction, a transesophageal echocardiography is more sensitive than a transthoracic echocardiography in detecting mitral regurgitation.

Definition of fibronectin-mediated uptake of gelatinized latex by liver slices and macrophages.

These studies show that both liver slices and macrophages carried out fibronectin concentration-dependent uptake of 125I-labeled gelatin-coated latex (test latex). Lack of phagocytosis of test latex by liver slices was shown directly by electron microscopy and indirectly by trypsin treatment, which caused the release of all test latex taken up in response to fibronectin. Inhibitors of phagocytosis did not alter this uptake. On the other hand, trypsin released only a portion of test latex from macrophages. Inhibitors of phagocytosis did not effect the released radioactive particles from macrophages but greatly reduced the trypsin-resistant radioactivity, taken as representing phagocytized particles. Opsonization of test latex with fibronectin did not require heparin but its association with liver slices occurred only in the presence of heparin. Macrophages, however, readily bound and internalized the opsonized test latex and heparin only potentiated these reactions. Gelatin competed with test latex for fibronectin for opsonization, but did not inhibit binding and phagocytosis of fibronectin-test latex complexes. Finally, soluble fibronectin-gelatin complexes did not compete for binding and phagocytosis of fibronectin-test latex complexes. Thus, fibronectin concentrated on the surface of latex is preferred for interaction with the fibronectin receptor of macrophages. Gelatin, however, was not essential for this reaction, because fibronectin directly coupled to latex was also readily taken up.

Evidence for the recycling nature of the fibronectin receptor of macrophages.

Plasma fibronectin (pFN) has been shown to mediate phagocytosis of several types of artificial particles and tissue debris by macrophages. In the present investigation some of the dynamic aspects of this receptor-mediated cellular process have been studied. Plasma fibronectin did not bind specifically to fibronectin (FN)-receptors of rat peritoneal macrophages at either 4 degrees C or 37 degrees C. On the other hand, pFN aggregated on the surface of gelatin-coated latex beads (gLtx) and 125I-labeled pFN covalently coupled to latex beads (pFN-Ltx) bound strongly to macrophages at both temperatures. Both of these particles were also internalized at 37 degrees C. Treatment of macrophages by chymotrypsin, thermolysin, or trypsin in a protein-free tissue culture medium did not affect either of the above reactions; however, pronase treatment strongly reduced both the binding and internalization of the pFN-coated particles. The pronase-treated macrophage monolayers in time regained their ability to bind and internalize pFN-gLtx when incubated in fresh tissue culture medium. Such recovery, however, did not take place when the medium contained cycloheximide. On the other hand, phagocytosis of pFN-gLtx was not affected directly by cycloheximide with untreated macrophages; this suggests that the FN-receptor recycles during sustained phagocytosis. This assumption was substantiated by the observations that some of the established lysosomotropic amines--i.e., chloroquine, dansylcadaverine, and dimethyldansylcadaverine--caused total inhibition of internalization without affecting the binding of particles to macrophages. Furthermore, chloroquine protected the FN-receptors against destruction by pronase. Together these results suggest that macrophage receptors for FN are protein, present both on the cell surface and intracellularly, and recycle between the plasma membrane and intracellular sites during phagocytosis.

Further studies on the modulation of blood coagulation by human serum amyloid P component and its acute phase homologue C-reactive protein.

Serum amyloid P component (SAP), and its acute phase homologue C-reactive protein (CRP), prolonged activated partial thromboplastin times (APTT) in cell free plasma when assayed at physiological concentrations in the presence of heparin. SAP also inhibited clot formation initiated through the extrinsic and terminal phases of coagulation in heparinized cell free plasma, an activity not shared with CRP. When CRP and SAP were similarly evaluated in whole blood using the thromboelastograph (TEG), CRP delayed the onset of coagulation and the initial rate of fibrin formation/polymerization; final clot patency was unaltered. SAP suppressed the anticoagulant activity of heparin in the TEG assay, unlike results obtained in heparinized cell free plasma, by facilitating a more rapid onset of coagulation, increasing the rate of fibrin formation/polymerization, and correcting clot patency. The data provided offer further evidence that these homologues can intercede in blood coagulation.