Protective Immunity against Listeria monocytogenes in Rats, Provided by HCl- and NaOH-Induced Listeria monocytogenes Bacterial Ghosts (LMGs) as Vaccine Candidates.

Listeria monocytogenes (Lm) bacterial ghosts (LMGs) were produced by the minimum inhibitory concentration (MIC) of HCl, H2SO4, and NaOH. Acid and alkali effects on the LMGs were compared by in vitro and in vivo analyses. Scanning electron microscope showed that all chemicals form lysis pores on the Lm cell envelopes. Real-time qPCR revealed a complete absence of genomic DNA in HCl- and H2SO4-induced LMGs but not in NaOH-induced LMGs. HCl-, H2SO4- and NaOH-induced LMGs showed weaker or missing protein bands on SDS-PAGE gel when compared to wild-type Lm. Murine macrophages exposed to the HCl-induced LMGs showed higher cell viability than those exposed to NaOH-induced LMGs or wild-type Lm. The maximum level of cytokine expression (TNF-α, iNOS, IFN-γ, and IL-10 mRNA) was observed in the macrophages exposed to NaOH-induced LMGs, while that of IL-1ß mRNA was observed in the macrophages exposed to HCl-induced LMGs. To investigate LMGs as a vaccine candidate, mice were divided into PBS buffer-injected, HCl- and NaOH-induced LMGs immunized groups. Mice vaccinated with HCl- and NOH-induced LMGs, respectively, significantly increased in specific IgG antibodies, bactericidal activities of serum, and CD4+ and CD8+ T-cell population. Antigenic Lm proteins reacted with antisera against HCl- and NOH-induced LMGs, respectively. Bacterial loads in HCl- and NaOH-induced LMGs immunized mice were significantly lower than PBS-injected mice after virulent Lm challenges. It suggested that vaccination with LMGs induces both humoral and cell-mediated immune responses and protects against virulent challenges.

Development of a Kit for Rapid Immunochromatographic Detection of Sacbrood Virus Infecting Apis cerana (AcSBV) Based on Polyclonal and Monoclonal Antibodies Raised against Recombinant VP1 and VP2 Expressed in Escherichia coli.

A Korean isolate of the sacbrood virus infecting Apis cerana (AcSBV-Kor) is the most destructive honeybee virus, causing serious economic damage losses in Korean apiculture. To address this, here, we attempted to develop an assay for the rapid detection of AcSBV-Kor based on immunochromatographic detection of constituent viral proteins. Genes encoding VP1 and VP2 proteins of AcSBV-Kor were cloned into an expression vector (pET-28a) and expressed in Escherichia coli BL21(DE3). During purification, recombinant VP1 (rVP1) and VP2 (rVP2) proteins were found in the insoluble fraction, with a molecular size of 26.7 and 24.9 kDa, respectively. BALB/c mice immunized with the purified rVP1 and rVP2 produced polyclonal antibodies (pAbs) such as pAb-rVP1 and pAb-rVP2. Western blot analysis showed that pAb-rVP1 strongly reacted with the homologous rVP1 but weakly reacted with heterologous rVP2. However, pAb-rVP2 strongly reacted not only with the homologous rVP2 but also with the heterologous rVP1. Spleen cells of the immunized mice fused with SP2/0-Ag14 myeloma cells produced monoclonal antibodies (mAbs) such as mAb-rVP1-1 and mAb-rVP2-13. Western blot analysis indicated that pAb-rVP1, pAb-rVP2, mAb-rVP1-1, and mAb-rVP2-13 reacted with AcSBV-infected honeybees and larvae as well as the corresponding recombinant proteins. These antibodies were then used in the development of a rapid immunochromatography (IC) strip assay kit with colloidal gold coupled to pAb-rVP1 and pAb-rVP2 at the conjugate pad and mAb-rVP1-1 and mAb-rVP2-13 at the test line. One antibody pair, pAb-rVP1/mAb-VP1-1, showed positive reactivity as low as 1.38 × 103 copies, while the other pair, pAb-rVP2/mAb-VP2-13, showed positive reactivity as low as 1.38 × 104 copies. Therefore, the antibody pair pAb-rVP1/mAb-VP1-1 was selected as a final candidate for validation. To validate the detection of AcSBV, the IC strip tests were conducted with 50 positive and 50 negative samples and compared with real-time PCR tests. The results confirm that the developed IC assay is a sufficiently sensitive and specific detection method for user-friendly and rapid detection of AcSBV.

Topical embryonic stem cells enhance wound healing in diabetic rats.

The effects of embryonic stem cells (ESCs) on diabetic wound healing were investigated using an excisional skin wound model in 110 diabetes-induced rats. We transplanted a clonal population of ESCs (5 × 10(6)) by topical injection into full thickness skin wounds. Four study groups were used; nondiabetic rats as a control, non-insulin controlled diabetic rats not treated with ESCs, insulin controlled diabetic rats not treated with ESCs, and insulin controlled diabetic rats treated with ESCs. Five rats in each experimental group were sacrificed on days 1, 5, 10, 15, and 20 after wounding. Wounds images were acquired daily and wound sizes were calculated. We measured the mRNA levels of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF), and fibronectin levels in extracellular matrix, and assessed wound healing by assessing histological parameters of epidermal regeneration, granulation tissue thickness, and angiogenesis. In the ESC-treated group, wound sizes were significantly smaller than in the insulin controlled diabetic group not treated with ESCs on days 5 and 10 (p < 0.05), and EGF and VEGF levels were markedly higher on days 5 and 10, fibronectin levels on day 5 after injection. All histological scores in the ESC-treated group were significantly higher than those of the insulin controlled diabetic group on day 5 (p < 0.05). Our results shows that topical ESCs enhance diabetic wound healing during the early stage, and suggest that ESCs transplantation offers a novel therapeutic modality for the treatment of diabetic wounds.

Patient-related factors and complications after arthroscopic tenotomy of the long head of the biceps tendon.

BACKGROUND: Tenotomy of the long head of the biceps tendon (LHBT) has been reported to provide reliable pain relief and require little postoperative rehabilitation. Complications such as cosmetic deformity, decrease in elbow flexion strength, decrease in supination strength, and fatigue discomfort have been reported after tenotomy of the LHBT. PURPOSE: To evaluate the complications of arthroscopic tenotomy of the LHBT in the shoulder-specifically, cosmetic deformity, decreased elbow flexion strength, and a cramplike arm pain-and to identify the patient-related factors that affect the rate of complications after tenotomy, such as, age, sex, involvement of the dominant arm, and body mass index (BMI). STUDY DESIGN: Case series; Level of evidence, 4. METHODS: In sum, 132 patients were evaluated. They had a mean age of 63 years (range, 26 to 82 years) at the time of surgery and a mean follow-up of 21 months (range, 12 to 53 months). The presence of so-called Popeye deformity, a cramplike arm pain at resisted elbow flexion, and strength of elbow flexion were evaluated. The development of each complication was compared by age, sex, involvement of the dominant arm, and BMI. RESULTS: Of the 132 patients, 60 (45%) had Popeye deformity, and it was significantly more frequent in men (76%) than women (31%) (P < .001). However, statistical analysis showed no difference in Popeye deformity frequency by age, arm dominance, or BMI. Male sex was found to be the only risk factor associated with the development of Popeye deformity, with an odds ratio of 10.21 versus women (95% confidence interval, 3.97 to 26.27; P < .001). Ten patients (8%) complained of a cramplike arm pain. Elbow flexion strength decreased in 60 patients (45%), although no intergroup differences were found for elbow flexion strength. CONCLUSION: The current study showed a 45% prevalence of Popeye deformity and 8% cramplike arm pain on exertion after tenotomy of the LHBT. Among patient factors such as sex, age, dominant arm relation, and body mass index, the male sex was the only factor correlated with occurrence of a Popeye deformity. Other factors did not show any correlation with deformity, elbow flexion strength, and cramplike arm pain.

Comparison of the outcomes of distraction osteogenesis for first and fourth brachymetatarsia.

BACKGROUND: Brachymetatarsia is the presence of an abnormally short metatarsal, and distraction osteogenesis has been used for its treatment. The purpose of the present study was to compare the outcomes of patients who underwent distraction osteogenesis for the treatment of first and/or fourth brachymetatarsia. METHODS: The data from forty-eight patients (sixty-four feet, seventy-four metatarsals) who underwent distraction osteogenesis for the treatment of brachymetatarsia were reviewed. The indications for surgery included a metatarsal that was at least 10 mm shorter than the adjacent metatarsal and that had an unacceptable cosmetic appearance. The study group comprised thirty-two first brachymetatarsia in nineteen patients (Group A) and forty-two fourth brachymetatarsia in twenty-nine patients (Group B). The average age was twenty years in Group A and eighteen years in Group B. The average duration of follow-up was 58.1 months in Group A and 56.1 in Group B. Metatarsal length, lengthening gain, healing time and index, and the American Orthopaedic Foot and Ankle Society hallux metatarsophalangeal-interphalangeal and lesser metatarsophalangeal-interphalangeal scores were evaluated. RESULTS: All patients were satisfied with the final length of the metatarsal and all had achieved bone union at the time of the last follow-up. The mean lengthening gain was 17.2 mm (42.9%) in Group A and 16.3 mm (37.3%) in Group B. The mean healing index was 71.0 days/cm in Group A and 67.3 days/cm in Group B. The mean American Orthopaedic Foot and Ankle Society score was 91.2 points in Group A and 92.8 points in Group B at the last follow-up (twenty-two excellent, nine good, and one fair results in Group A, and twenty-nine excellent, eleven good, and two fair results in Group B). The most common complication was metatarsophalangeal joint stiffness, which occurred in thirteen rays in Group A and in twelve rays in Group B; malalignment of the lengthened metatarsal was observed six times in each group. No significant intergroup differences in the outcomes were found. CONCLUSIONS: Distraction osteogenesis for first and/or fourth brachymetatarsia provided successful lengthening of a metatarsal with eventual osseous union and was associated with similar outcomes in terms of healing index, function score, and the prevalence of complications between the two groups, although frequent complications were encountered and no improvement in foot function was found.

Prospective evaluation of the effectiveness of a home-based program of isometric strengthening exercises: 12-month follow-up.

BACKGROUND: The aim of this prospective randomized clinical trial was to investigate the efficacy of a home-based program of isometric strengthening exercises for the treatment of the lateral epicondylitis (LE) of the distal humerus. We hypothesized that 1) use of isometric strengthening exercises would result in clinical benefits similar to those provided by medication and pain relief and 2) functional improvements after exercise would be time-dependent. METHODS: Patients were assigned to one of two groups: 1) an immediate physical therapy group (group I), or 2) a delayed physical therapy group (group D). Group I patients (n = 16) were instructed how to do the exercises at their first clinic visit and immediately carried out the exercise program. Group D patients (n = 15) learned and did the exercises after being on medications for 4 weeks. RESULTS: Outcomes at the 1-month clinic visit indicated that pain (measured using a visual analogue scale [VAS]) had been significantly reduced in group I compared to group D (p < 0.01). However, significant differences between groups were not found at 3-, 6-, and 12-month follow-up for either VAS scores or Mayo elbow performance scores. For modified Nirschl/Pettrone scores, a significant difference between groups was found only at the 1-month follow-up visit. By then, the number of participants who returned to all activities with no pain or occasional mild pain was six (37%) in Group I and two (13%) in Group D (p = 0.031). At the final follow-up visit, 88% of all participants performed physical activities without pain. CONCLUSIONS: Isometric strengthening exercises done early in the course of LE (within 4 weeks) provides a clinically significant improvement.

How to avoid complications of distraction osteogenesis for first brachymetatarsia.

BACKGROUND AND PURPOSE: Distraction osteogenesis may be used for the treatment of brachymetatarsia. However, few reports have been published on first metatarsal lengthening by this method. We evaluated the complications of distraction osteogenesis for first brachymetatarsia and here we provide a solution. PATIENTS AND METHODS: 16 patients (27 feet) underwent distraction osteogenesis for first brachymetatarsia. Mean age at time of surgery was 20 (12-34) years and mean duration of postoperative follow-up was 5 (2-13) years. A unilateral external fixator was fixed at the medial aspect of the metatarsus. The distraction axis was parallel to the plantar surface of the foot in the sagittal plane and to the second metatarsal axis in the transversal plane. RESULTS: First metatarsal length expressed as a proportion of second metatarsal length was 60% (55-64). Average degree of metatarsal lengthening was 42% (34-54), and the average lengthening index was 64 (39-93) days/cm. The most common complication was stiffness of the metatarsophalangeal joint (12 feet). Deformities that included cavus foot and hallux valgus occurred in 3 feet each, and callus fractures occurred in 3 feet. The other complications were pin breakage and pin tract infection in 2 feet each. INTERPRETATION: Distraction osteogenesis for first brachymetatarsia can give satisfactory cosmetic and functional results. However, several complications are commonly encountered. This report on complications and their solutions may help those attempting distraction osteogenesis for first brachymetatarsia.

Surgical treatment of pseudosyndactyly of children with epidermolysis bullosa: a case report.

Recessive dystrophic epidermolysis bullosa (RDEB) is an uncommon and severe disorder characterised by trauma-induced blisters, intractable skin ulcers, scarring, milia, and nail dystrophy. We report the good result of both surgical release of fingers allowing spontaneous epithelisation without skin grafting and post-operative meticulous skin care without splinting, followed-up for one year.

Expression and significance of TGF-beta isoform and VEGF in osteosarcoma.

This article demonstrates that the degree of VEG expression could be used as a clinically signific prognostic factor in osteosarcoma.

Residual shortening after Legg-Calve-Perthes disease, focusing on the response of the ipsilateral tibia.

Residual shortening of the affected limb was measured at skeletal maturity by teleoroentgenograms in 68 patients with Legg-Calve-Perthes disease (LCPD); special attention was paid to the length of the ipsilateral tibia. Of these 68 patients, 38 were treated by abduction orthosis (AO) and 30 by femoral varus osteotomy (FVO). Residual shortening in AO group was significantly greater than that in FVO group. The femoral lengths in both of these groups were similar (12.5 mm in the AO group and 10.1 mm in the FVO group), but the tibial lengths were significantly different (2.5 mm shortening in the AO group and 0.9 mm lengthening in the FVO group). Residual shortening in the patients treated by FVO was less than that in patients treated by AO. The difference is speculated to be caused by the overgrowth of the ipsilateral tibia.

Dysplasia epiphysealis capitis femoris: Meyer dysplasia.

The clinical importance of dysplasia epiphysealis capitis femoris (Meyer dysplasia) is that it is easily mistaken for Legg-Calve-Perthes disease, leading to unnecessary diagnostic procedures and treatments. After a review of 578 children (619 hips) with Legg-Calve-Perthes disease, 17 children (27 hips) in whom both the clinical and radiologic pattern was obviously different could be found and a diagnosis of dysplasia epiphysealis capitis femoris was finally made. The mean age was 2.5 (range 1.9-3.6) years. There were 16 boys and 1 girl. Ten children had bilateral involvement (59%). The capital femoral epiphysis was delayed or was smaller in 26 hips, separated or cracked in 15, and cystic in 6. A normal bone structure was established in approximately 2 to 4 years. The final results assessed by the Mose and the Stulberg classification were good in all 27 hips. This study suggests guidelines for evaluating this rare condition based on the authors' findings and a review of the literature.

Subcutaneous emphysema mimicking gas gangrene following perforation of the rectum: a case report.

We report a case of extensive subcutaneous emphysema of the lower extremity mimicking gas gangrene following perforation of the rectum in a 38-yr-old man. Subcutaneous emphysema of the leg may rarely occur secondary to perforation of the gastrointestinal tract and has often created serious diagnostic problems and high mortality rates. Therefore, prompt diagnosis and aggressive treatment is imperative.

Failure of osteointegration of hamstring tendon autograft after anterior cruciate ligament reconstruction.

Anterior cruciate ligament (ACL) reconstruction is completed after implantation, when the graft material used undergoes extensive biologic remodelling and osteointegration. Failure of the osteointegration between the graft tendon and bone decreases the graft strength and induces anterior instability. We experienced 2 cases of failed osteointegration between tendon and bone after ACL reconstruction using 4 strands of the hamstring tendon. Surprisingly, osteointegration between the bone and tendon junction was not shown by intraoperative findings. The histologic findings also presented no evidence of osteointegration or biologic remodeling of the tendon. Two cases of failure of osteointegration are reported, with a review of the literature.

Fate of the osteochondral fragments in osteochondritis dissecans after Legg-Calve-Perthes' disease.

The treatment of osteochondritis dissecans after Legg-Calvé-Perthes' disease has not been clearly determined. It may be either by simple observation or surgical removal of the osteochondral fragment. We studied the evolution of the lesion in 13 children and reviewed 92 hips reported in the literature. In our patients ten showed a tendency towards spontaneous healing, one required drilling + grafting to obtain fusion, and in two there was separation into the joint. These loose bodies were in the acetabular fossa and caused no symptoms. On reviewing the literature, we found only four cases of hips with loose bodies from osteochondritis dissecans. These were lying in the inferomedial capsule and were also asymptomatic. Treatment of osteochondritis dissecans after Legg-Calvé-Perthes' disease should therefore be conservative unless the fragment interferes with the mechanics of the hip.