Production and cell surface display of recombinant anthrax protective antigen on the surface layer of attenuated Bacillus anthracis.

To investigate the surface display of the anthrax protective antigen (PA) on attenuated Bacillus anthracis, a recombinant B. anthracis strain, named AP429 was constructed by integrating into the chromosome a translational fusion harboring the DNA fragments encoding the cell wall-targeting domain of the S-layer protein EA1 and the anthrax PA. Crerecombinase action at the loxP sites excised the antibiotic marker. Western blot analysis, fluorescence-activated cell sorting and immunofluorescence analysis confirmed that PA was successfully expressed on the S-layer of the recombinant antibiotic marker-free strain. Notwithstanding extensive proteolytic degradation of the hybrid protein SLHs-PA, quantitative ELISA revealed that approximately 8.1 × 10(6) molecules of SLHs-PA were gained from each Bacillus cell. Moreover, electron microscopy assay indicated that the typical S-layer structures could be clearly observed from the recombinant strain micrographs.

A walk-in screening of dementia in the general population in Taiwan.

Alzheimer's disease (AD) has increased in its prevalence due to the increasing aged population. Currently there is no updated data on the prevalence of dementia including its very mild stage in Taiwan. Under the extensive coverage of Mentality Protection Center (MPC), Fo Guang Shan, Taiwan, the volunteers of MPC have conducted the medicine-related services and the screening of dementia by AD8 (ascertainment of dementia 8) that can screen the dementia even at its very mild stage in general population in all Taiwan. From 2011 to 2013, in total, 2,171 participants, 368 in the northern, 549 in the central, 877 in the southern, and 377 in the eastern part, were recruited with the mean age being 66.9 ± 10.2 years old. The ratio of suspected dementia patients, AD8 score greater than or equal to 2, was 13.6% of all recruited participants with their mean AD8 score being 2.9 ± 1.3, mean age being 69.4 ± 10.8 years old, and female predominance being 73.0%. Although this is a screening study, it has extensive coverage of all Taiwan and the use of AD8 is capable of screening very mild dementia. A further study with a randomized sampling to examine the prevalence and incidence of dementia including its very mild stage is encouraged.

A community-based walk-in screening of depression in Taiwan.

Depression is a crucial public health problem because of its relatively high association with suicidal attempts, prolonged social isolation, poor physical health, and dementia. However, the available data and study on the prevalence of depression in Taiwan were mostly completed within the previous 1 to 2 decades, and these studies were limited to certain areas or populations. Little is known regarding the current status of depression in Taiwan. We used a brief tool, the Center for Epidemiological Studies Depression Scale (CES-D), to screen depression in 4 areas among the general and aged population. The results showed a higher CES-D score in the southern area among general (mean ± SD: 7.8 ± 8.4) or aged participants (mean ± SD: 7.2 ± 8.0) compared with other areas. The ratio of suspected depression patients was 16.4% of all recruited participants and 13.3% of aged participants. These results may provide information for this public health issue.

Comparison of the Post-Surgical Position of the Temporomandibular Joint after Orthognathic Surgery in Skeletal Class III Patients and Patients with Cleft Lip and Palate.

OBJECTIVE: The purpose of our research is to compare the post-surgical position of the temporomandibular joint in skeletal Class III patients and patients with cleft lip and palate treated with two-jaw orthognathic surgery using a three-dimensional computer tomography image. MATERIALS AND METHODS: Twenty-three skeletal Class III patients with mandibular prognathism associated with maxillary retrognathism in group 1 and twenty cleft mid-face retrusion skeletal Class III patients in group 2 were enrolled in this study. All subjects were treated with two-jaw orthognathic surgery. Computed tomography scans were taken in all subjects at 3 weeks preoperatively and 6 months postoperatively. Three-dimensional craniofacial skeletal structures were build-up, and assessed the temporomandibular joint position changes before and after surgery. RESULTS: Forty-three selected patients were separated into two groups. The mean age of patients was 22.39 ± 4.8 years in group 1 and 20.25 ± 3.8 years in group 2. The range of mean three-dimensional discrepancy of the selected condylar points was 0.95-1.23 mm in group 1 and 2.37-2.86 mm in group 2. The mean alteration of intercondylar angle was 2.33 ± 1.34° in group 1 and 6.30 ± 2.22° in group 2. The significant differences in the discrepancy of TMJ and changes in intercondylar angle were confirmed within the intra-group and between the two groups. CONCLUSIONS: Significant changes in postoperative TMJ position were present in both groups. Furthermore, the cleft group presented significantly more postoperative discrepancy of TMJ and more changes in intercondylar angle after surgery. This finding may be a reason leading to greater postoperative instability in cleft patients compared with skeletal Class III non-cleft patients. CLINICAL TRIAL REGISTRATION NUMBER: IRB No: 202201108B0.

Angiotensin-converting enzyme gene and plasma protein level in Alzheimer's disease in Taiwanese.

BACKGROUND: angiotensin-converting enzyme (ACE) gene insertion/deletion (indel) polymorphism is considered a biomarker for Alzheimer's disease (AD). However, the associations of ACE gene and protein level to AD are undetermined among Taiwanese. METHODS: this study investigated 257 Taiwanese cases with AD and 137 ethnically matched controls using ACE gene indel genotype association methods with logistic regression adjusted for other variables. Besides, 65 out of 257 AD patients, 11 with D/D genotype, 28 with I/I genotype and 26 with I/D genotype were recruited. Their plasma ACE protein levels were measured by enzyme-linked immuno-sorbent assay and compared for their corresponding ACE gene indel polymorphism. RESULTS: patients with ACE-I/I homozygote were less likely to be associated with AD, compared with both I/D and D/D (OR: 0.601; 95% CI: 0.372-0.969; P = 0.037), or only I/D genotype (OR: 0.584; 95% CI: 0.349-0.976; P = 0.040). There were significantly different plasma ACE protein levels among these three different genotype groups (P = 0.023). The I/I genotype group had significantly lower ACE plasma levels [114.79 ± 31.32 ng/ml (mean ± SD)], compared with D/D (164.07 ± 86.36 ng/ml; P = 0.010), but not I/D (141.45 ± 51.50 ng/ml; P = 0.064). CONCLUSION: ACE-I/I homozygote corresponds to lower plasma ACE protein level and it is independently but less likely to be associated with AD. These findings signal the importance of ACE indel polymorphisms to their corresponding protein levels and to AD.

Postsurgical Stability of Temporomandibular Joint of Skeletal Class III Patients Treated with 2-Jaw Orthognathic Surgery via Computer-Aided Three-Dimensional Simulation and Navigation in Orthognathic Surgery (CASNOS).

OBJECTIVE: The aim of this study is to clarify the postsurgical stability of temporomandibular joints in skeletal class III patients treated with 2-jaw orthognathic surgery which was performed utilizing computer-aided three-dimensional simulation and navigation in orthognathic surgery (CASNOS) protocol. MATERIALS AND METHODS: 23 consecutive nongrowing skeletal class III patients with mandibular prognathism associated with maxillary retrognathism treated with 2-jaw orthognathic surgery between 2018 and 2019 were enrolled in this study. The surgery was planned according to the standardized protocol of CASNOS (computer-aided three-dimensional simulation and navigation in orthognathic surgery). Computed tomography (CT) scans were performed in all patients 3 weeks presurgically and 6 months postsurgically. ITKSNAP and 3D Slicer software were used to reconstruct three-dimensional facial skeletal images, to carry out image segmentation, and to superimpose and quantify the TMJ position changes before and after surgery. Amount of displacement of the most medial and lateral points of the condyles and the change of intercondylar angles were measured to evaluate the postsurgical stability of TMJ. RESULTS: A total amount of 23 skeletal class III patients (female : male = 12 : 11) with age ranged from 20.3 to 33.5 years (mean: 24.39 ± 4.8 years old) underwent Le Fort I maxillary advancement and BSSO setback of the mandible. The surgical outcome revealed the satisfactory correction of their skeletal deformities. The mean displacement of the right most lateral condylar point (RL-RL') was 1.04 ± 0.42 mm and the mean displacement of the left most lateral condylar point (LL-LL') was 1.19 ± 0.41 mm. The mean displacement of the right most medial condylar point (RM-RM') was 1.03 ± 0.39 mm and the left most medial condylar point (LM-LM') was 0.96 ± 0.39 mm. The mean intercondylar angle was 161.61 ± 5.08° presurgically and 159.28 ± 4.92° postsurgically. CONCLUSION: The postsurgical position of TM joint condyles in our study only presented a mild change with all the landmark displacement within a range of 1.2 mm. This indicates the bimaxillary orthognathic surgery via 3D CASNOS protocol can achieve a desired and stable result of TMJ position in treating skeletal class III adult patients with retrognathic maxilla and prognathic mandible.

Construction of a novel Shigella live-vector strain co-expressing CS3 and LTB/STm of enterotoxigenic E.coli.

AIM: To construct and evaluate a polyvalent recombinant vaccine strain Shigella flexneri 2a T32 against enterotoxigenic E.coli (ETEC). METHODS: By using a host-plasmid balanced lethal system based on asd gene, a polyvalent recombinant strain was constructed to highly express CS3 and regularly express fusion enterotoxin of LTB subunit and mutant ST (LTB/STm) in a vaccine strain Shigella flexneri 2a T32 with specific deletion of asd gene. Fimbria CS3 was observed by immunofluorescence and electron microscopy assay. The security of LTB/STm was examined by ileal loop assay and suckling mouse assay. To evaluate this new candidate vaccine, it was compared with a previous vaccine strain in plasmid and protein level, growth assay and immunogenicity in Balb/c mice. RESULTS: The newly constructed vaccine expressed CS3 and grew better than the previously constructed vaccine except for the lower expression of LTB/STm. Serum IgG and mucosal IgA against CS3, LTB, ST, and host lipopolysaccharide (LPS) were produced after immunization of Balb/c mice by oral route with the new strain. The titers were not significantly different from the Balb/c mice with the previous strain. CONCLUSION: This novel candidate diarrheal vaccine can effectively induce serum and mucosal antibody responses against ETEC and Shigella.

[Progress on the vaccine for anthrax].

Bacillus anthracis is the causative organism of the potentially fatal disease anthrax, and the used vaccines have some disadvantages. There are new developments appeared for the Bacillus anthracis in recent years, such as anti-PA antibody kills the spore of Bacillus anthracis, mucosal immunization induces immune responses in both systemic and secretory immune compartments, Poly (gamma-D-PGA) protein induce IgG antibodies to the vegetative bacteria, new pathogens were found by genomic analysis. The DNA vaccine and live vector vaccine will be the next generation vaccines for anthrax. It will have a shorter immunization schedule and will be greater protective efficacy than before.

Effect of music intervention on the cognitive and depression status of senior apartment residents in Taiwan.

OBJECTIVE: To identify the effect of music intervention on cognitive function and depression status of residents in senior citizen apartments based on the existing evidence regarding music therapy. METHODS: An experimental study was conducted from November 2008 to December 2009. Sixty healthy senior apartment residents over 65 years of age were recruited and separated into two groups. According to their opinion, 41 took part in the music intervention group and 19 in the comparison group. The music intervention involved Buddhist hymns. The short-term effects were evaluated based on the measurement of cognitive function and depression level using the Mini-Mental State Examination (MMSE) and the Geriatric Depression Scale-short form (GDS-SF) at the baseline, 1 month, and 4 months. RESULTS: The means of the initial and the 1-month MMSE and GDS-SF scores did not differ between the two groups. The 4-month MMSE score significantly declined compared with the initial level in the comparison group, whereas no significant change was observed in the experimental group. Moreover, the 4-month GDS-SF score significantly improved in both groups compared with the initial level. CONCLUSION: Music intervention may postpone cognitive decline in healthy residents preferring Buddhist hymns in the senior citizen apartments in 4 months follow-up, and intense contact with participants may improve their mood status.

[Co-expression of CFA/I and CS6 of Enterotoxigenic Escherichia coli ( ETEC ) in Shigella flexneri 2a T32 Derivative Strain FWL01].

Among the known colonization factors of enterotoxigenic Escherichia coli (ETEC), CFA/I and CS6 (the common antigen in the CFA/IV fimbrial antigens ) are two of the most prevalent fimbriae found in clinical isolates but are never expressed by the same wild-type strains. In this study, CFA/I and CS6 of ETEC were co-expressed in Shigella flexneri 2a T32 derivative strain FWL01 by using a host-plasmid lethal balancing system based on asd gene. The results indicate that the recombinant plasmid carrying CFA/I and CS6 could be stably integrated in FWL01. Expression of the two antigens did not interfere the host growth. The results of immunofluorescence analysis showed that CFA/I and CS6 were localized on the surface of the strain FWL01. In Balb/c mice orally immunized with the recombinant strain, the immune responses against CFA/I and CS6 were observed. Those observations show the feasibility of a multivalent vaccine expressing different fimbrial antigens in attenuated Shigella flexneri.

[Clinical result of forefoot correction by the first ray stabilization combined with resection of the lesser metatarsal head procedure for patient with rheumatoid arthritis].

OBJECTIVE: To introduce the procedure of the 1st ray stabilization combined with resection of the lesser metatarsal heads for patient with severe forefoot deformity caused by rheumatoid arthritis (RA) and evaluate the short to mid-term clinical results. METHODS: From Oct. 2006 to Aug. 2010,97 patients (129 feet) aged from 36 to 67 years (average 54), with forefoot deformity caused by rheumatoid arthritis were reviwed. There were 88 males and 9 females,65 single lateral involved and 32 bilateral involved, the average duration of disease was 17 years (6 to 32 years). The 1st ray instability and lesser metatarsophalangeal (MTP) joint stiff dislocation were found in all cases. The first ray stabilization combined with resection of the lesser metatarsal head procedure were performed for all cases. The radiographic Hallux valgus angle (HVA) and intermetatarsal angle (IMA) were measurde and the JSSF (Japanese Society for Surgery of the Foot) score were evaluated before operation and every follow up. RESULTS: The average followed-up was 37 months (6 to 52 months) for all patients except 5 (7 feet) and 1 died for acute cardiac infarction 1 year after operation. The average JSSF score improved from (33.2 +/- 8.2) points preoperative to (67.3 +/- 3.1) points at final followed-up (P < 0.01); the average HVA was corrected from (50.0 +/- 11.8) degrees preoperative to (21.2 +/- 3.2) degrees at final follow up (P < 0.01); the average IMA was corrected from (15.5 +/- 3.6) degrees preoperative to (9.7 +/- 6.6) degrees at final follow up (P < 0.01). MTP joint nonunion was found in 4 feet. A radiographic high density mass was found in the 1st cuneiform bone during 8 to 11 months followed-up in 3 feet; delayed wound healing was happened in 9 feet; MTP joint infection was happened in 2 feet; tarsometatarsal joint infection was happened in 1 foot; lesser MTP joints deformity recurrence were found in 16 feet. CONCLUSION: The characters of forefoot with RA in later stage are the 1st ray deformity and instability compound with the lesser toes deformity. The 1st ray stability procedure which include the 1st MTP arthrodesis and the Lapidus procedure can correct the 1st ray deformities and rebuilt its stability. The lesser toes metatarsal head resection is effective in correct their deformity. This combined procedure is reliable. It is suitable for patients with severe Hallux valgus, increased IMA, tarsometatarsal joint instability and the lesser MTP joint stiff dislocation.

Association of NOD1 and NOD2 genes polymorphisms with Helicobacter pylori related gastric cancer in a Chinese population.

AIM: To investigate the association between the tag single nucleotide polymorphisms (TagSNPs) of NOD1 and NOD2 and the risk of developing gastric cancer. METHODS: We conducted a hospital-based case-control study including 296 incident gastric cancer patients and 160 gastritis controls. Eight TagSNPs in the NOD1 and NOD2 genes were selected from the Hapmap database using the haploview software and genotyped by the Sequenom MassArray system. The serum levels of anti-Helicobacter pylori (H. pylori) IgG were measured by enzyme-linked immunosorbent assay to indicate H. pylori infection. The odds ratios (OR) and 95% confidence intervals (CI) were calculated by unconditional logistic regression, including sex and age as confounding factors. RESULTS: The NOD1 rs2907749 GG genotype showed a decreased risk for gastric cancer (OR 0.50, 95% CI: 0.26-0.95, P = 0.04) while the rs7789045 TT genotype showed an increased risk (OR 2.14, 95% CI: 1.20-3.82, P = 0.01). An elevated susceptibility to gastric cancer was observed in the subjects with H. pylori infection and the NaOD1 rs7789045 TT genotype (OR 2.05, 95% CI: 1.07-3.94, P = 0.03) or the NOD2 rs7205423 GC genotype (OR 2.52, 95% CI: 1.05-6.04, P = 0.04). Haplotype analysis suggested that the distribution of AGT (rs2907749, rs2075820 and rs7789045) in NOD1 between the cases and control groups was significantly different (P corrected: 0.04), and the diplotype AGT/AGT was associated with an elevated gastric cancer risk (OR 1.98, 95% CI: 1.04-3.79, P = 0.04). The association of the NOD1 rs7789045 TT genotype and the diplotype AGT/AGT was significant with H. pylori-related diffuse-type gastric cancer (OR 3.00, 95% CI: 1.38-6.53, P = 0.01; OR 4.02, 95% CI: 1.61-10.05, P < 0.01, respectively). CONCLUSION: Genetic polymorphisms in NOD1 and NOD2 may interact with H. pylori infection and may play important roles in promoting the development of gastric cancer in the Chinese population.

Identification of B-cell epitopes in urease B subunit of Helicobacter pylori bound by neutralizing antibodies.

To identify linear B-cell epitopes of urease B (UreB), a series of 19 partially overlapping fragments of the UreB gene were expressed. Three MAbs against UreB of Helicobacter pylori (H. pylori), A1H10, A3C10, and B3D9, were tested for their reactivity to the truncated proteins by Western blot and enzyme-linked immunosorbent assay (ELISA). Three linear B-cell epitopes were identified covering a stretch of 15 amino acid (aa) residues and localized in the aa regions 158-172, 181-195, and 349-363 of UreB. ELISA also showed that the three synthetic peptides containing epitope sequences (UP32: GGGTGPADGTNATTI, UP35: WMLRAAEEYSMNLGF, and UP38: TLHDMGIFSITSSDS) were recognized by the corresponding MAbs and H. pylori positive sera from H. pylori infected patients. Mice immunized with glutathione S-transferase (GST) fusion peptides showed that epitope-specific antibodies were capable of inhibiting urease enzymatic activity. These results should be useful in clinical applications and highlight the potential importance of these epitopes as the targets for development of epitope-based vaccines against H. pylori.