Reliability and validity of surface EMG assessments combined with isometric muscle strength testing in patients with abdominal rectus diastasis and asymptomatic controls.

PURPOSE: Patients with abdominal rectus diastasis (ARD) may have muscular functional impairments, but clinics lack appropriate objective assessment tools. The aim was to establish the relative and absolute reliability, and convergent validity, of muscular activity using Surface Electromyography (SEMG) during isometric abdominal muscle strength testing in patients with ARD and controls without ARD. METHODS: Twenty-six patients with ARD were matched for age, sex and BMI with controls without ARD. Participants were tested twice during isometric muscular contractions using SEMG located on six abdominal sites. Mean amplitude, fatigue, and recruitment order were analyzed. Relative reliability was evaluated with Intraclass Correlation Coefficients (ICC), while absolute reliability was estimated by calculating the Standard Error of Measurement and Minimal Detectable Change. Convergent validity was addressed in relation to participant characteristics, functional ability, and symptoms. RESULTS: Mean SEMG amplitude for all abdominal wall muscle contractions showed moderate to excellent relative test-retest reliability, with ICC values ranging from 0.46 to 0.97. In contrast, fatigue and recruitment order displayed poor to moderate relative reliability in both groups. Absolute reliability measures were generally high. A moderate to high convergent validity (ARD: rho-value 0.41-0.70; Controls: rho-value 0.41-0.75) was observed for mean amplitude in relation to a functional sit-to-stand test, abdominal circumference, BMI, back pain, and quality-of-life. CONCLUSIONS: The results of applying SEMG during isometric abdominal muscle support practicing the method in clinics, although additional development is needed with further standardization and more functional testing. Furthermore, the method demonstrates construct validity in patients with ARD and in age- and sex-matched controls.

The Sheep Grimace Scale as an indicator of post-operative distress and pain in laboratory sheep.

The EU Directive 2010/63/EU changed the requirements regarding the use of laboratory animals and raised important issues related to assessing the severity of all procedures undertaken on laboratory animals. However, quantifiable parameters to assess severity are rare, and improved assessment strategies need to be developed. Hence, a Sheep Grimace Scale (SGS) was herein established by observing and interpreting sheep facial expressions as a consequence of pain and distress following unilateral tibia osteotomy. The animals were clinically investigated and scored five days before surgery and at 1, 3, 7, 10, 14 and 17 days afterwards. Additionally, cortisol levels in the saliva of the sheep were determined at the respective time points. For the SGS, video recording was performed, and pictures of the sheep were randomized and scored by blinded observers. Osteotomy in sheep resulted in an increased clinical severity score from days 1 to 17 post-surgery and elevated salivary cortisol levels one day post-surgery. An analysis of facial expressions revealed a significantly increased SGS on the day of surgery until day 3 post-surgery; this elevated level was sustained until day 17. Clinical severity and SGS scores correlated positively with a Pearson´s correlation coefficient of 0.47. Further investigations regarding the applicability of the SGS revealed a high inter-observer reliability with an intraclass correlation coefficient of 0.92 and an accuracy of 68.2%. In conclusion, the SGS represents a valuable approach for severity assessment that may help support and refine a widely used welfare assessment for sheep during experimental procedures, thereby meeting legislation requirements and minimizing the occurrence of unrecognized distress in animal experimentation.

Bacteriome and Mycobiome Interactions Underscore Microbial Dysbiosis in Familial Crohn's Disease.

UNLABELLED: Crohn's disease (CD) results from a complex interplay between host genetic factors and endogenous microbial communities. In the current study, we used Ion Torrent sequencing to characterize the gut bacterial microbiota (bacteriome) and fungal community (mycobiome) in patients with CD and their nondiseased first-degree relatives (NCDR) in 9 familial clusters living in northern France-Belgium and in healthy individuals from 4 families living in the same area (non-CD unrelated [NCDU]). Principal component, diversity, and abundance analyses were conducted, and CD-associated inter- and intrakingdom microbial correlations were determined. Significant microbial interactions were identified and validated using single- and mixed-species biofilms. CD and NCDR groups clustered together in the mycobiome but not in the bacteriome. Microbiotas of familial (CD and NCDR) samples were distinct from those of nonfamilial (NCDU) samples. The abundance of Serratia marcescens and Escherichia coli was elevated in CD patients, while that of beneficial bacteria was decreased. The abundance of the fungus Candida tropicalis was significantly higher in CD than in NCDR (P = 0.003) samples and positively correlated with levels of anti-Saccharomyces cerevisiae antibodies (ASCA). The abundance of C. tropicalis was positively correlated with S. marcescens and E. coli, suggesting that these organisms interact in the gut. The mass and thickness of triple-species (C. tropicalis plus S. marcescens plus E. coli) biofilm were significantly greater than those of single- and double-species biofilms. C. tropicalis biofilms comprised blastospores, while double- and triple-species biofilms were enriched in hyphae. S. marcescens used fimbriae to coaggregate or attach with C. tropicalis/E. coli, while E. coli was closely apposed with C. tropicalis Specific interkingdom microbial interactions may be key determinants in CD. IMPORTANCE: Here, we characterized the gut bacterial microbiota (bacteriome) and fungal community (mycobiome) in multiplex families with CD and healthy relatives and defined the microbial interactions leading to dysbiosis in CD. We identified fungal (Candida tropicalis) and bacterial (Serratia marcescens and Escherichia coli) species that are associated with CD dysbiosis. Additionally, we found that the level of anti-Saccharomyces cerevisiae antibodies (ASCA; a known CD biomarker) was associated with the abundance of C. tropicalis We also identified positive interkingdom correlations between C. tropicalis, E. coli, and S. marcescens in CD patients and validated these correlations using in vitro biofilms. These results provide insight into the roles of bacteria and fungi in CD and may lead to the development of novel treatment approaches and diagnostic assays.

Iron-cofactored superoxide dismutase inhibits host responses to Mycobacterium tuberculosis.

Superoxide dismutase (SOD) is a ubiquitous metalloenzyme in aerobic organisms that catalyzes the conversion of superoxide anion to hydrogen peroxide. Mycobacterium tuberculosis is unusual in that it secretes large quantities of iron-cofactored SOD. To determine the role of SOD in pathogenesis, we constructed mutants of M. tuberculosis H37Rv with reduced SOD production. Compared with controls, SOD-diminished isolates were more susceptible to killing by hydrogen peroxide. The isolates were markedly attenuated, exhibiting nearly 100,000-fold fewer bacilli than virulent control strains in the lungs and spleens of C57BL/6 mice 4 wk after intravenous inoculation. In the lung, SOD-attenuated M. tuberculosis induced robust interstitial mononuclear cell infiltration within 24 h and many cells were apoptotic by TUNEL staining, whereas virulent H37Rv exhibited minimal early inflammatory response and only rare interstitial mononuclear cell apoptosis. During prolonged infections, C57BL/6 mice tolerated SOD-attenuated M. tuberculosis better than BCG, exhibiting 68% greater weight gain, quicker eradication of bacilli from the spleen, and less alveolar lung infiltration. These results establish the importance of SOD in the pathogenesis of tuberculosis. Its effect appears to be mediated in part by inhibiting innate host immune responses, including early mononuclear cell infiltration of infected tissues and apoptosis.

Expression of a retinoid-inducible tumor suppressor, Tazarotene-inducible gene-3, is decreased in psoriasis and skin cancer.

Tazarotene-induced gene-3 (TIG-3), isolated from human keratinocytes treated with the retinoic acid receptor-selective retinoid Tazarotene, is homologous to H-rev, a class II tumor suppressor. TIG-3 gene localized to chromosome 11q23, a site of loss of heterozygosity in several malignancies. Retinoids influence epidermal differentiation and are used to treat and prevent skin cancer. Therefore, we studied TIG-3 mRNA expression in psoriasis and in basal and SCCs by in situ hybridization and a quantitative QT-RT-PCR assay. Psoriasis lesions had significantly lower staining (median, 3) than paired normal control skin (median, 4; P = 0.012). TIG-3 mRNA was significantly higher in normal control skin (P = 0.001), in paired adjacent skin (median, 3; P = 0.007), and in overlying epidermis (median, 3.0; P = 0.0001) than in 21 SCC specimens as a group (median, 1.5).

Cutaneous lesions of metastatic visceral malignancy mimicking pyogenic granuloma.

Cutaneous metastases may be the first sign of a previously undiagnosed visceral malignancy or the initial presentation of a recurrent neoplasm. Rarely, skin metastases can resemble a pyogenic granuloma. Three oncology patients who developed new pyogenic granuloma-like cutaneous lesions are described. Histopathologic examination showed metastatic visceral malignancy in the skin. The characteristics of the previously reported cancer patients with metastatic tumor to the skin that mimicked a pyogenic granuloma are reviewed. A biopsy of a skin lesion that clinically appears to represent a pyogenic granuloma should be performed for microscopic examination in patients with a previous visceral malignancy or in cancer-free individuals whose lesions do not resolve after conservative treatment.

Management of localized scleroderma.

Localized scleroderma denotes a spectrum of conditions characterized by circumscribed fibrotic areas involving different levels of the dermis, subcutis, and sometimes underlying soft tissue and bone. Although the clinical course of the disease is often benign, widespread lesions and disabling joint contractures may lead to significant complications. The pathogenesis of the different types of localized scleroderma is still unknown. Numerous therapeutic agents have been reported to be effective in this disease spectrum, but controlled studies are rare. The purpose of this review is to summarize previous experience and to discuss recent advances in the management of localized scleroderma.

Bath-PUVA therapy in three patients with scleredema adultorum.

BACKGROUND: Scleredema adultorum (SA) is a rare connective tissue disorder for which no treatment has proven to be effective. OBJECTIVE: Our purpose was to determine the effect of bath-PUVA therapy on SA. METHODS: Three patients were treated. Clinical evaluation of skin induration and thickness as well as ultrasonography were performed at baseline and after treatment. RESULTS: All three patients showed substantial clinical improvement with bath-PUVA therapy (median of 59 treatments and a cumulative UVA dose of 245.7 J/cm2). Ultrasonography showed significant reduction in both skin thickness and density. CONCLUSION: Bath-PUVA therapy appears to be effective in the treatment of SA.

Segmental neurofibromatosis: case reports and review.

Segmental neurofibromatosis (neurofibromatosis type V) is a rare disorder characterized by café-lu-lait macules and neurofibromas, or only neurofibromas, limited to one region of the body. Three patients with segmental neurofibromatosis are described, and cases of this condition in the world literature are reviewed. Segmental neurofibromatosis has only been described in 82 patients, including our three. The median age at onset was 28 years and the incidence was higher in women (58%). The neurofibromas most commonly occupied either a cervical or thoracic dermatome and were unilateral, occurring more often on the right side (43 patients) than the left (34 patients). Café-au-lait macules were present in 26% of patients. Axillary freckling was described in only nine patients. Disease-associated systemic involvement was uncommon. Most patients with segmental neurofibromatosis (93%) do not have a family history of neurofibromatosis.

SP-A enhances uptake of bacillus Calmette-Guérin by macrophages through a specific SP-A receptor.

Surfactant-associated protein A (SP-A) is a C-type lectin that is involved in surfactant metabolism as well as host defense functions in the lung. We have recently identified a receptor on macrophages [specific 210-kDa SP-A receptor (SPR210)] that binds SP-A. In the current study we have investigated the role of SP-A in mediating uptake of bacillus Calmette-Guérin (BCG) by rat macrophages and human monocytes and have examined the role of the macrophage SPR210 in this process. 125I-labeled SP-A bound BCG in a Ca(2+)-, carbohydrate-, and dose-dependent manner. To examine association of SP-A-BCG complexes with macrophages, BCG were opsonized with SP-A and were incubated with rat bone marrow-derived macrophages (RBMM), rat alveolar macrophages (RAM), or human monocytes at a 1-to-1 ratio for 4 h. The cells were washed, fixed in formalin, and stained with auramine-rhodamine. Cell-associated organisms were enumerated by fluorescent microscopy. The percentage of cells with one or more associated BCG was increased by SP-A from 27% of RBMM with BCG alone to 54% with SP-A-BCG complexes; 1-16% in RAM; and 39-67% in human monocytes. This enhanced uptake was dependent on the dose of SP-A, with maximal increases seen with 10 micrograms/ml. Electron microscopic analysis supported the conclusion that organisms were ingested by and not simply bound to the macrophages. Inclusion of SPR210 antibodies blocked association of SP-A-BCG complexes, suggesting a role for SPR210 in mediating the interaction of SP-A-BCG with the macrophages. This was further supported by the finding that modulation of SPR210 activity resulted in altered SP-A-BCG uptake. These results demonstrate that SP-A binds to BCG and that uptake of these SP-A-BCG complexes is mediated in part by the SPR210 on rat macrophages and human monocytes.

Clinical and functional outcome of anterior cruciate ligament reconstruction in the recreational athlete over the age of 35.

This study examined the functional, objective, and subjective outcome of anterior cruciate ligament (ACL) reconstruction in recreational athletes > or = 35 years after a minimum of 2 years of follow-up. Patients > or = 35 years who underwent ACL reconstruction by a single surgeon were identified from our surgical database. Nineteen knees in 18 (62% follow-up) patients were available for review by an independent examiner. The patients underwent physical examination, radiographs, functional testing, isokinetic strength testing, and instrumented ligament arthrometer testing. All were seen at a minimum of 2 years of follow-up. The average age was 40 years. Five of 19 underwent reconstruction less than 1 month after injury, and the remainder underwent reconstruction for chronic injuries. All patients preoperatively had at least a grade 2 Lachman and a positive pivot shift noted on physical examination. After a minimum of 2 years of follow-up, 17 of 18 patients had a stable knee on objective testing, including a negative Lachman and pivot shift. Seventeen patients (94%) had < 3 mm side-to-side difference on maximum manual arthrometric testing. Only one patient had > 3 cm prone heel height difference, and all patients had > 125 degrees of flexion. Mean thigh circumference difference was 0.5 cm. Isokinetic testing demonstrated a mean 11%, 7%, and 4% quadriceps asymmetry at 60 degrees, 180 degrees, and 240 degrees/second, respectively. However, functional testing revealed only a mean 6% asymmetry on vertical jump, single leg hop, and timed 6 meter hop. Seventeen of 18 patients were satisfied with their results. The mean postoperative Lysholm Rating Scale score was 93. The mean Noyes Sports Activity Scale score was 86, improved from 31 preoperatively. Thirteen of 18 returned to their preinjury level of sports performance. These results indicate that ACL reconstruction in patients over the age of 35 has functional, objective, and subjective results comparable to those of a younger patient population.

Arthrometric results of arthroscopically assisted anterior cruciate ligament reconstruction using autograft patellar tendon substitution.

Sixty-two patients who underwent arthroscopically assisted anterior cruciate ligament reconstruction with the middle-third patellar tendon autograft without extraarticular augmentation were retrospectively reviewed at a minimum 2-year (mean, 37-month) followup. A postoperative KT-1000 arthrometric evaluation revealed a mean maximum manual difference of 0.3 mm (range, -6 to +16). Ninety percent of the patients had a mean maximum difference of < or = 3 mm. Three patients had a > or = 5 mm mean maximum difference; positive pivot shifts were noted in two of these patients. Despite an early range of motion, early weightbearing protocol without an extraarticular backup, the pivot shift (92% negative) was reliably eliminated at postoperative followup. Arthrometric parameters were statistically reduced (P < 0.0001) from preoperative status and were consistent with the diagnostic criteria established for normal knees. There was no relationship established in the "tighter" knees (those with negative side-to-side arthrometric differences) and the incidence of knee flexion contractures, patellar pain symptoms, Cybex extension deficit (> 20%), functional indices, or postoperative rating scales.

Surgical resection of the distal clavicle.

Distal clavicle resection is frequently performed for persistent acromioclavicular pain. However, patient outcome after this procedure has not been well described. The results of subjective and objective retrospective analysis of 23 open distal clavicle resections in 21 patients are reported. Patients were evaluated with a questionnaire, roentgenographs, physical examination, and isokinetic muscle strength testing at an average of 30 months after surgery. Eighteen of 23 shoulders had good or excellent postoperative ratings. All patients had normal motion. No significant weakness of the operated shoulder was seen on objective muscle testing when compared with the unoperated shoulder. The average preoperative Hospital for Special Surgery score was 34, and the average postoperative score was 84. Age and hand dominance were unrelated to postoperative outcomes. Less predictable results were obtained in patients with workmen's compensation and in those patients involved in litigation. Open distal clavicle resection yields good to excellent results in properly selected patients and does not create significant subjective or objective weakness.

Arthroscopy-assisted anterior cruciate ligament reconstruction using patellar tendon substitution. Two- to four-year follow-up results.

The purpose of this retrospective study was to evaluate clinically, functionally, and objectively our initial experience using free, autogenous middle third patellar tendon for anterior cruciate ligament reconstruction without extraarticular augmentation in 62 of 75 patients (80% followup) who were available for clinical review at a minimum 2-year followup. Subjective, clinical, functional, Cybex dynamometer, and KT-1000 arthrometer tests were performed along with modified tests of the Hospital for Special Surgery, Noyes Cincinnati, Tegner, and Lysholm knee rating scales. Ninety-two percent had a negative pivot shift at followup. The mean Cybex dynamometer extension deficits postoperatively were 9% and 7% at 180 and 240 deg/sec. Mild patellar pain symptoms were noted in 18%. The reoperation rate was 10% with a mild flexion contracture as the most common reason. The Hospital for Special Surgery scoring scale postoperatively was 88; Noyes, 86; Lysholm, 88; and Tegner, 6. Mean postoperative single-legged and vertical jump indices were 88% and 87%, respectively. The KT-1000 arthrometric evaluation postoperatively revealed a mean maximum manual difference of 0.3 mm; 92% of the patients had a maximum manual difference of < or = 3 mm. Subjectively, 95% indicated that they would undergo the procedure again. Early results demonstrate excellent stability, preservation of motion, and encouraging evaluations by scoring scales and arthrometric evaluation.

Detection of Rochalimaea henselae DNA in specimens from cat scratch disease patients by PCR.

A PCR assay was developed by using degenerate primers that allow amplification of a 414-bp fragment of DNA from the rickettsia-like organisms Rochalimaea henselae and R. quintana. Internal oligonucleotides were used as hybridization probes, permitting rapid differentiation of these two Rochalimaea species. DNAs from 12 different isolates of R. henselae were amplified with the PCR primers, and the resulting 414-bp PCR product hybridized only with the R. henselae-specific probe. DNAs from four different isolates of R. quintana were amplified and produced a PCR product of the same size that hybridized only with the R. quintana-specific probe. DNAs from isolates of R. elizabethae, R. vinsonii, Bartonella bacilliformis, and Afipia felis failed to amplify the 414-bp fragment in the PCR assay. This two-step assay was applied to DNAs extracted from 16 fresh (unfixed) lymph node biopsy specimens and nine aspirates from patients with clinical cat scratch disease (CSD) to assay for the presence of R. henselae or R. quintana DNA in these samples. Twenty-one (84%) of 25 lymph node samples from CSD patients were positive for R. henselae, while none were positive for R. quintana. The characteristic 414-bp fragment was not amplified from eight lymph node tissue samples from non-CSD cases. These results provide evidence that R. henselae, and not R. quintana, plays the central role in the etiology of CSD.

Identification and characterization of two nonessential regions of the rabbitpox virus genome involved in virulence.

We have developed a means to identify genes associated with particular aspects of virulence. By beginning with an avirulent deletion mutant of rabbitpox virus and systematically reintroducing overlapping segments of the deleted region, we have identified two regions of the viral genome associated with increased virulence in mice. Evaluation of illness has been aided by the exploitation of weight loss as an indicator of pathogenesis. One of the regions identified by this method contains several open reading frames and includes two previously described genes. A third, as yet undescribed, gene within this region potentially encodes a product related to the C5 protein of human complement. The second region of DNA associated with increased virulence is the HindIII M fragment, which contains only one complete open reading frame. Analysis of this previously unreported gene shows coding potential for a polypeptide of 254 amino acids (approximately 25 kDa) which is related to the C4 component of human complement. The elucidation of two new viral genes related to complement components, taken together with the recent report of the biological activity of a poxvirus-encoded complement-binding protein, suggests the importance of interactions of the virus with the complement system during a normal infection.

An antimelanoma-barley ribosome inactivating protein conjugate is cytotoxic to melanoma cells in vitro.

The ribosome inactivating protein (BRIP) from barley is a single polypeptide chain (Mr = 32,000 Dalton) and is nontoxic to intact cells. The BRIP has been purified to homogeneity by modifications of the methods of Roberts and Selitrennikoff and crosslinked to monoclonal antibodies by Succinimidyl-3 (2- Pyridyldithio) -Propionate (SPDP) and by the cystamine-EDAC methods. The resulting hybrids were purified from the free BRIP by gel filtration on a Sephadex G-75 column. The model suicide transport agents were assayed against melanoma cells; K-562 cells were used as control. The hybrids were found to be selectively toxic to melanoma cells in a dose dependent manner.

A possible role for protein kinase C activity but not cyclic nucleotide-dependent protein kinases in human natural killer cell lytic activity.

Recent evidence suggests that a Ca++, phospholipid, diacylglycerol-dependent protein kinase, protein kinase C, plays a role in the activation of cytotoxic T lymphocytes by target cells. In this investigation we have examined the role of protein kinase C in human NK cell-mediated cytolysis of K-562 cells. The protein kinase C inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) inhibited human NK cell-mediated cytolysis in a dose dependent manner. On the other hand, N-(2-guanidinoethyl)-5-isoquinolinesulfonamide (HA1004), a specific inhibitor of cyclic nucleotide dependent protein kinases had no effect on human NK cell-mediated cytolysis of K562 cells. There is little or no effect on protein synthesis or N-glycosylation activity in human NK cells by H-7. The relative inhibitory ability of the two inhibitors suggest that protein kinase C, acting synergistically with Ca++ mobilization, plays a role in the early stages of human NK cell-mediated cytolysis of K562 target cells.