The function and affinity maturation of HIV-1 gp120-specific monoclonal antibodies derived from colostral B cells.

Despite the risk of transmitting HIV-1, mothers in resource-poor areas are encouraged to breastfeed their infants because of beneficial immunologic and nutritional factors in milk. Interestingly, in the absence of antiretroviral prophylaxis, the overwhelming majority of HIV-1-exposed, breastfeeding infants are naturally protected from infection. To understand the role of HIV-1 envelope (Env)-specific antibodies in breast milk in natural protection against infant virus transmission, we produced 19 HIV-1 Env-specific monoclonal antibodies (mAbs) isolated from colostrum B cells of HIV-1-infected mothers and investigated their specificity, evolution, and anti-HIV-1 functions. Despite the previously reported genetic compartmentalization and gp120-specific bias of colostrum HIV Env-specific B cells, the colostrum Env-specific mAbs described here demonstrated a broad range of gp120 epitope specificities and functions, including inhibition of epithelial cell binding and dendritic cell-mediated virus transfer, neutralization, and antibody-dependent cellular cytotoxicity. We also identified divergent patterns of colostrum Env-specific B-cell lineage evolution with respect to crossreactivity to gastrointestinal commensal bacteria, indicating that commensal bacterial antigens play a role in shaping the local breast milk immunoglobulin G (IgG) repertoire. Maternal vaccine strategies to specifically target this breast milk B-cell population may be necessary to achieve safe breastfeeding for all HIV-1-exposed infants.

Restricted isotype, distinct variable gene usage, and high rate of gp120 specificity of HIV-1 envelope-specific B cells in colostrum compared with those in blood of HIV-1-infected, lactating African women.

A successful HIV-1 vaccine must elicit immune responses that impede mucosal virus transmission, though functional roles of protective HIV-1 Envelope (Env)-specific mucosal antibodies remain unclear. Colostrum is a rich source of readily accessible mucosal B cells that may help define the mucosal antibody response contributing to prevention of postnatal HIV-1 transmission. To examine the HIV-1 Env-specific colostrum B-cell repertoire, single B cells were isolated from 17 chronically HIV-infected, lactating women, producing 51 blood and 39 colostrum HIV-1 Env-specific B-cell antibodies. All HIV-1 Env-specific colostrum-derived antibodies were immunoglobulin (Ig)G1 isotype and had mean heavy chain complementarity-determining region 3 (CDR3) lengths and mutation frequencies similar to those isolated from blood. However, variable heavy chain (VH) gene subfamily 1(∼)69 usage was higher among colostrum than blood HIV-1 Env-reactive antibodies (49% vs. 20%, P=0.006, Fisher's exact test). Additionally, more HIV-1 Env-specific colostrum antibodies were gp120 specific than those isolated from blood (44% vs. 16%, P=0.005, Fisher's exact test). One cross-compartment HIV-1 Env-specific clonal B-cell lineage was identified. These unique characteristics of colostrum B-cell antibodies suggest selective homing of HIV-1-specific IgG1-secreting memory B cells to the mammary gland and have implications for targeting mucosal B-cell populations by vaccination.

A CMOS wireless biomolecular sensing system-on-chip based on polysilicon nanowire technology.

As developments of modern societies, an on-field and personalized diagnosis has become important for disease prevention and proper treatment. To address this need, in this work, a polysilicon nanowire (poly-Si NW) based biosensor system-on-chip (bio-SSoC) is designed and fabricated by a 0.35 µm 2-Poly-4-Metal (2P4M) complementary metal-oxide-semiconductor (CMOS) process provided by a commercialized semiconductor foundry. Because of the advantages of CMOS system-on-chip (SoC) technologies, the poly-Si NW biosensor is integrated with a chopper differential-difference amplifier (DDA) based analog-front-end (AFE), a successive approximation analog-to-digital converter (SAR ADC), and a microcontroller to have better sensing capabilities than a traditional Si NW discrete measuring system. In addition, an on-off key (OOK) wireless transceiver is also integrated to form a wireless bio-SSoC technology. This is pioneering work to harness the momentum of CMOS integrated technology into emerging bio-diagnosis technologies. This integrated technology is experimentally examined to have a label-free and low-concentration biomolecular detection for both Hepatitis B Virus DNA (10 fM) and cardiac troponin I protein (3.2 pM). Based on this work, the implemented wireless bio-SSoC has demonstrated a good biomolecular sensing characteristic and a potential for low-cost and mobile applications. As a consequence, this developed technology can be a promising candidate for on-field and personalized applications in biomedical diagnosis.

Proliferation and osteo/odontoblastic differentiation of stem cells from dental apical papilla in mineralization-inducing medium containing additional KH(2)PO(4).

OBJECTIVES: Stem cells from the dental apical papilla (SCAPs) can be induced to differentiate along both osteoblast and odontoblast lineages. However, little knowledge is available concerning their differentiation efficiency in osteogenic media containing additional KH2 PO4 . MATERIALS AND METHODS: Stem cells from the dental apical papilla were isolated from apical papillae of immature third molars and treated with two kinds of mineralization-inducing media, MM1 and MM2, differing in KH2 PO4 concentration. Proliferation and osteo/odontogenic differentiation capacity of MM1/MM2-treated SCAPs were investigated and compared both in vitro and in vivo. RESULTS: Cell counting and flow cytometry demonstrated that MM2 containing 1.8 mm additional KH2 PO4 significantly enhanced proliferative potential of SCAPs, compared to MM1. Osteo/odontogenic capacity of SCAPs was much better in MM2 medium than in MM1, as indicated by elevated alkaline phosphatase activity, increased calcium deposition and upregulated expression of osteo/odontoblast-specific genes/proteins (for example, runt-related transcription factor 2, osterix, osteocalcin, dentin sialoprotein, and dentin sialophosphoprotein). In vivo transplantation findings proved that SCAPs in MM2 group generated more mineralized tissues, and presented higher expression of osteo/odontoblast-specific proteins (osteocalcin and dentin sialoprotein) than those in the MM1 group. CONCLUSION: Mineralization-inducing media supplemented with 1.8 mm additional KH2 PO4 significantly enhanced cell proliferation and improved differentiation capacity of SCAPs along osteo/odontogenic cell lineages, compared to counterparts lacking additional KH2 PO4 .

Effects of variety, sowing season and maturity stage on ruminal phosphorus release from whole maize stover and its morphological fractions.

An in situ degradation technique was used to investigate the effects of variety, season and maturity stage on ruminal Phosphorus (P) release from whole maize stover and morphological fractions from five varieties of maize (conventional maize, CM; sweet maize, SM; waxy maize, WM; high oil maize, HOM; and fodder maize, FM). Maize plants were harvested in 2005, 17 and 31 days after tasselling and manually separated into leaf blade, leaf sheath, stem and husk. Results showed that the values of rapidly released P fractions (a), slowly released P fractions (b), rate constant of P release for fraction b (c) and effective disappearance (ED) of P were significantly influenced by variety, sowing season and maturity stage (p < 0.05). The ED of P in whole maize stover among the varieties decreased in the following order: FM (89.8%) > HOM (87.9%) > CM (87.0%) > SM (86.9%) > WM (83.9%). Advanced maturity stage increased (p < 0.001) the a and ED values of P in the leaf blade and leaf sheath, but decreased (p < 0.001) these parameters in the husk. The a fraction and ED of P were higher (p < 0.001) for maize sown in spring than those sown in summer. Significant (p < 0.05) interactions among variety, sowing season and maturity stage were noted in the a and ED values of P. Effective disappearance of morphological fraction varied significantly (p < 0.001) and followed the order: stem (90.3%) > leaf sheath (88.2%) > leaf blade (85.2%) > husk (84.9%). In conclusion, most of P in maize stover could be released in the rumen. The potential pollution resulting from P excretion in faeces may be alleviated, when maize stover is largely used as the main forage source of ruminants.

Effect of phosphorus availability on basal root shallowness in common bean.

Root gravitropism may be an important element of plant response to phosphorus availability because it determines root foraging in fertile topsoil horizons, and thereby phosphorus acquisition. In this study we seek to test this hypothesis in both two dimensional paper growth pouch and three-dimensional solid media of sand and soil cultures. Five common bean ( Phaseolus vulgaris L.) genotypes with contrasting adaptation to low phosphorus availability were evaluated in growth pouches over 6 days of growth, and in sand culture and soil culture over 4 weeks of growth. In all three media, phosphorus availability regulated the gravitropic response of basal roots in a genotype-dependent manner. In pouches, sand, and soil, the phosphorus-inefficient genotype DOR 364 had deeper roots with phosphorus stress, whereas the phosphorus-efficient genotype G19833 responded to phosphorus stress by producing shallower roots. Genotypes were most responsive to phosphorus stress in sand culture, where relative root allocation to the 0-3- and 3-6-cm horizons increased 50% with phosphorus stress, and varied 300% (3-6 cm) to 500% (0-3 cm) among genotypes. Our results indicate that (1) phosphorus availability regulates root gravitropic growth in both paper and solid media, (2) responses observed in young seedlings continue throughout vegetative growth, (3) the response of root gravitropism to phosphorus availability varies among genotypes, and (4) genotypic adaptation to low phosphorus availability is correlated with the ability to allocate roots to shallow soil horizons under phosphorus stress.

Isolation and characterization of an active compound from black soybean [Glycine max (L.) Merr.] and its effect on proliferation and differentiation of human leukemic U937 cells.

Black soybean [Glycine max (L.) Merr.] has been used as a health food and herb in China for hundreds of years. In the present study, we purified a unique polysaccharide component from black soybean (PSBS) and found that it indirectly inhibits proliferation and induces differentiation of human leukemic U937 cells via activation of mononuclear cells (MNCs). We prepared conditioned media (MNC-CM) by incubating MNCs from human peripheral blood with or without PSBS (PSBS-MNC-CM and normal MNC-CM, respectively). Treatment of human leukemic U937 cells with PSBS-MNC-CM significantly inhibited proliferation of U937 cells, reducing their growth by 98.5%. Furthermore, PSBS-MNC-CM induced U937 cells to differentiate into mature monocytes/macrophages (83% by morphological examination and 90% by the nitroblue tetrazolium test). Neither PSBS alone nor normal MNC-CM had such effects. The molecular weight of PSBS was about 480 000 by gel filtration. Structural analysis of PSBS revealed that (1,6)-alpha-D-glucan might be its major active component. Our results suggest that the PSBS may inhibit proliferation and induce differentiation in human leukemic U937 cells by activating the immune response of MNCs.

Adrenal insufficiency in critically ill emergency department patients: a Taiwan preliminary study.

OBJECTIVE: Unrecognized adrenal insufficiency can have serious consequences in critically ill emergency department (ED) patients. This prospective pilot study of adrenal function in patients with severe illness was undertaken to determine the prevalence of adrenal dysfunction and any relation to prior herbal drug use. METHODS: In a high-volume urban tertiary care ED, adult patients with sepsis or acute myocardial infarction (AMI) were eligible for the study. Over a two-month period, a convenience sample was enrolled by the authors on arrival to the ED. Inclusion criteria were systemic inflammatory response syndrome (SIRS) criteria plus evidence of at least one organ dysfunction or cardiac marker plus electrocardiogram-proven AMI. Exclusion criteria included known corticosteroid use. Serum cortisol was measured on arrival and for those patients with a level of <15 microg/dL (<414 nmol/L), an adrenocorticotropic hormone (ACTH) stimulation test was performed. RESULTS: Of the 30 enrolled patients, 23 (77%) were suffering from severe sepsis and the other seven (23%) had an AMI. Thirteen of the 30 patients (43%; 95% CI = 25% to 65%) had serum cortisol levels of <15 microg/dL, consistent with adrenal insufficiency, nine with severe sepsis and four with an AMI. Eight (62%; 95% CI = 32% to 86%) of the 13 patients with low cortisol levels reported using herbal medications, while only two (12%; 95% CI = 1% to 36%) of the 17 with normal cortisol levels reported taking herb drugs (p = 0.01). Only two (15%; 95% CI = 2% to 45%) of the patients with low cortisol levels failed their corticotropin stimulation test, suggestive of true adrenocortical insufficiency. Both reported using herbal preparations. CONCLUSIONS: These results indicate that adrenal dysfunction is common among a group of critically ill patients seen in this Taiwanese ED. Moreover, the use of herbal drugs was high in the patients with low serum cortisols. Further studies are required to both confirm these findings and clarify whether a number of herbal medications contain corticosteroids.

Induction of a major leaf acid phosphatase does not confer adaptation to low phosphorus availability in common bean.

Acid phosphatase is believed to be important for phosphorus scavenging and remobilization in plants, but its role in plant adaptation to low phosphorus availability has not been critically evaluated. To address this issue, we compared acid phosphatase activity (APA) in leaves of common bean (Phaseolus vulgaris) in a phosphorus-inefficient genotype (DOR364), a phosphorus-efficient genotype (G19833), and their F(5.10) recombinant inbred lines (RILs). Phosphorus deficiency substantially increased leaf APA, but APA was much higher and more responsive to phosphorus availability in DOR364 than in G19833. Leaf APA segregated in the RILs, with two discrete groups having either high (mean = 1.71 micromol/mg protein/min) or low (0.36 micromol/mg protein/min) activity. A chi-square test indicated that the observed difference might be controlled by a single gene. Non-denaturing protein electrophoresis revealed that there are four visible isoforms responsible for total APA in common bean, and that the difference in APA between contrasting genotypes could be attributed to the existence of a single major isoform. Qualitative mapping of the APA trait and quantitative trait loci analysis with molecular markers indicated that a major gene contributing to APA is located on linkage group B03 of the unified common bean map. This locus was not associated with loci conferring phosphorus acquisition efficiency or phosphorus use efficiency. RILs contrasting for APA had similar phosphorus pools in old and young leaves under phosphorus stress, arguing against a role for APA in phosphorus remobilization. Our results do not support a major role for leaf APA induction in regulating plant adaptation to phosphorus deficiency.

Data compression for arterial pulse waveform.

The arterial pulse possesses important clinical information in traditional Chinese medicine. It is usually recorded for a long period of time in the applications of telemedicine and PACS systems. Due to the huge amount of data, by recognizing the strong correlation between successive beat patterns in arterial pulse waveform sequences, a novel and efficient data compression scheme based mainly on pattern matching is introduced. The simulation results show that our coding scheme can achieve a very high compression ratio and low distortion for arterial pulse waveform.

Effect of tooth-bound fluoride on enamel demineralization/ remineralization in vitro.

The effect of tooth-bound fluoride (F) on enamel caries formation was investigated under the condition that loosely bound F was essentially absent. Eighteen thin enamel sections, prepared from the lingual or buccal surfaces of extracted human molars, were embedded in acrylic resin with the enamel surfaces exposed. The sections were placed in a pH 7 remineralizing solution (RS; 1.2 mmol/l Ca, 0.72 mmol/l P, 30 mmol/l KCl, 50 mmol/l HEPES) for 5 days, and were randomly divided into 3 groups: (1) control group that received no treatment, (2) acidulated phosphate fluoride (APF) group that received 5 cycles of a 4 min treatment with APF gel followed by immersion in the RS for 2 days (RS changed daily) and (3) dicalcium phosphate dihydrate (DCPD) - APF group that received 5 cycles of a 4-min pH 2.1 DCPD-forming solution followed by 4 min APF gel and then placed in the RS for 2 days. After the treatment cycles, the sections were washed in a constant composition F titration system to remove loosely bound F. An in vitro model, which consisted of cycles of de- (6 h) and remineralization (18 h) each day for 5 days, was used to produce caries-like lesions in the specimens. The DeltaZ (mineral loss) values, measured by quantitative microradiography, of the lesions formed in the three groups were (mean +/- standard deviation; n = 6) 91.2+/-12.3 microm for the control group, 41.3+/-10.1 microm for the APF group and 21.2+/-4.8 microm for the DCPD-APF group. The same system produced lesions in untreated shark enamel with a mean DeltaZ of 4.4+/-0.3 microm (n = 12). One-way fixed-effects ANOVA indicated that mineral loss was significantly different among the different groups (p<0.05). The results showed that enamel resistance to lesion formation increased with increasing tooth-bound F content. Shark enamel was much more resistant to demineralization than human enamel.

[Wind-damage effects on quality of heartwood of Lignum Santali Albi].

OBJECTIVE: To evaluate the wind-damage effects on quality of heartwood of Lignum Santali Albi. METHOD: GC-MS, TLC and pharmacodynamic test. RESULTS: The content of volatile oil from heartwood of Wind-damaged Lignum Santali Albi is 1.42%; the content of various components in the oil and the chromatography of different extracts are similar to those of reference drug and 25 years old trees. CONCLUSION: Wind-damage should accelerate the formation of heartwood of Lignum Santali Albi without influence on its quality.

[Experimental study on pharmacological action of fukean tablet on gastrointestinal tract].

The pharmacological effects of FuKean Tablet(FKT) on gastrointestinal tract were studied. The results showed FKT cured the diarrhea induced by rhubarb or senna, prominently reduced the quantity of feces and delayed the time of excreting charcoal powder in the diarrhea induced by rhubarb or senna model NIH mice; FKT inhibited intestinal propelling-movement in diarrhea induced by senna mmodel NIH mice. FKT inhibited the automatic contraction and Ach induced spasmodic contraction of isolated rabbit intestine(decreased the frequency and intensity of contraction). The above results revealed that the pharmacological base of curing the urgent disease in gastrointestinal tract may be curing the diarrhea, inhibiting intestinal propelling-movement and inhibiting the contraction of intestine.

[Otoacoustic emissions for newborn hearing screening].

OBJECTIVE: To evaluate the validity of transient evoked otoacoustic emissions (TEOAE) and distortion product otoacoustic emissions (DPOAE) in newborn hearing screening and to establish the feasible pass/fail criteria of screening. METHODS: One hundred and eight newborns (216 ears) were screened by TEOAE and DPOAE with Celesta 503 Cochlear Emission Analyzer. The outcome of OAE screening was compared with that of auditory brainstem response(ABR) test. RESULTS: When the criterion of passing TEOAE screening was based on the correlation coefficient of waveforms equal to or greater than 0.60 (r > or = 0.60), the co-positivity and co-negativity to the ABR results were 100% and 93.8% respectively. When the pass criterion of DPOAE was set as that the response levels were greater than the 5th percentile (P5) of the normal newborns in more than five out of eight frequencies, the co-positivity and co-negativity were 100% and 94.3% respectively. The outcome of TEOAE and DPOAE screening between r > or = 0.60 and P5 showed the best correlation, r = 0.6583 (n = 216, P = 0.0001). CONCLUSION: r > or = 0.60 and P5 were the feasible pass criteria of TEOAE and DPOAE for newborn hearing screening respectively. OAE proved to be an objective tool for rapid and effective test in universal newborn hearing screening.

Antagonistic effects of extract from leaves of ginkgo biloba on glutamate neurotoxicity.

AIM: To determine whether the extract of leaves of Ginkgo biloba L (EGb) and several active constituents of EGb have protective effects against glutamate (Glu)-induced neuronal damage. METHODS: Microscopy and image analysis of nucleus areas in the arcuate nuclei (AN) of mice were made. The neuronal viability in primary cultures from mouse cerebral cortex was assessed using MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] staining and the intracellular free calcium concentration ([Ca2+]i) of single neuron was measured using Fura-2. RESULTS: EGb (2.5 mg.L-1) and its constituent ginkgolide B (Gin B, 2 mg.L-1) protected the neuronal viability against Glu-induced injury, and prevented the Glu-induced elevation in [Ca2+]i. EGb (3-10 mg.kg-1) attenuated the decrease of nucleus areas in arcuate nuclei induced by Glu (1 g.kg-1, s.c.). CONCLUSION: EGb and Gin B prevent neurons from Glu neurotoxicity through reduction of the rise in [Ca2+]i.

[Effect of Epimedium sagittatum on quality of life and cellular immunity in patients of hemodialysis maintenance].

Our previous study had demonstrated that Chinese medicine Epimedium Sagittatum (ES) exerted immuno-enhancing effect on the animal model of chronic renal insufficiency. In present study, we investigated the therapeutic effect of ES on patients of hemodialysis maintenance. 22 cases of regular hemodialytic patients were treated with ES in the form of decoction. 12 patients with hemodialysis were served as controls. It was found that ES had sexual potentiation effect and improved the quality of life in the patients of chronic renal failure with regular hemodialysis. Interleukin 2 (IL-2) activity of peripheral blood monocytes (PBMC) stimulated by PHA was increased significantly in the patients treated with ES. It was suggested that Chinese medicine ES had therapeutic effect on sexual disorder and immunologic inadequacy in the patients of chronic renal failure undergoing hemodialysis.

Polyphenols, astringency and proline-rich proteins.

Recent, NMR and precipitation, studies of molecular recognition of proline-rich proteins and peptides by plant polyphenols are described and rationalized. The action of polysaccharides and caseins in the moderation of the astringent response, which is engendered by polyphenols present in foodstuffs and beverages, is described. The possible influence of plant cell wall glycoproteins on the process of lignification is discussed in the light of the observed affinity of phenolic substrates for prolyl residues in protein structures.

[Inorganic element analysis on different compatibility of sini decoction].

Thirteen kinds of inorganic elements in Sini decoction with different compatibility were analysed by atomic absorption spectrophotometry. The change in quantity and kind of medicines has no obvious effect on each element. But the extraction rate is reduced according to the increase in quantity and herb medicines in the prescription.

Cardiac preservation is enhanced in a heterotopic rat transplant model by supplementing the nitric oxide pathway.

Nitric oxide (NO) is a novel biologic messenger with diverse effects but its role in organ transplantation remains poorly understood. Using a porphyrinic microsensor, the first direct measurements of coronary vascular and endocardial NO production were made. NO was measured directly in the effluent of preserved, heterotopically transplanted rat hearts stimulated with L-arginine and bradykinin; NO concentrations fell from 2.1 +/- 0.4 microM for freshly explanted hearts to 0.7 +/- 0.2 and 0.2 +/- 0.08 microM for hearts preserved for 19 and 38 h, respectively. NO levels were increased by SOD, suggesting a role for superoxide-mediated destruction of NO. Consistent with these data, addition of the NO donor nitroglycerin (NTG) to a balanced salt preservation solution enhanced graft survival in a time- and dose-dependent manner, with 92% of hearts supplemented with NTG surviving 12 h of preservation versus only 17% in its absence. NTG similarly enhanced preservation of hearts stored in University of Wisconsin solution, the clinical standard for preservation. Other stimulators of the NO pathway, including nitroprusside, L-arginine, or 8-bromoguanosine 3',5' monophosphate, also enhanced graft survival, whereas the competitive NO synthase antagonist NG-monomethyl-L-arginine was associated with poor preservation. Likely mechanisms whereby supplementation of the NO pathway enhanced preservation included increased blood flow to the reperfused graft and decreased graft leukostasis. NO was also measured in endothelial cells subjected to hypoxia/reoxygenation and detected based on its ability to inhibit thrombin-mediated platelet aggregation and serotonin release. NO became undetectable in endothelial cells exposed to hypoxia followed by reoxygenation and was restored to normoxic levels on addition of SOD. These studies suggest that the NO pathway fails during preservation/transplantation because of formation of oxygen free radicals during reperfusion, which quench available NO. Augmentation of NO/cGMP-dependent mechanisms enhances vascular function after ischemia and reperfusion and provides a new strategy for transplantation of vascular organs.