In situ detection of SOCS and cytokine expression in the uterine cervix from HIV/HPV coinfected women.

The purpose of this study was to look for associations between a newly described class of suppressors of cytokine signaling (SSI/SOCS) and cytokine expression in the uterine cervix from HIV/HPV coinfected women. We examined the pro-inflammatory cytokines TNF-alpha and IL-6 since their expressions are linked and responsible for many aspects of both localized and systemic inflammatory responses. Further, expression of SSI/SOCS has been implicated in the negative feedback regulation of cytokine receptor signaling. PCR-amplified HIV-1 cDNA was noted mainly in the stroma, showing a perivascular distribution, and most of the infected cells colabeled with the macrophage marker CD68. The distribution of IL-6 and TNF-alpha was in the same area to HIV-1 and much greater than normal cervices from women with no evidence of viral infection. SOCS/SSI-1 and -3 mRNA positive cells in the uterine cervix were commonly detected in these noninfected cervical tissues; however, very few cells that contained SOCS were evident in areas where HIV-1, TNF-alpha, and IL-6 expressing cells were found. This suggests that viral-related suppression of SOCS/SSI-1-3 expression may be a factor in the marked local enhancement of TNF-alpha and IL-6 production which, in turn, may help facilitate viral spread; however, further studies should be done in order to elucidate the exact mechanisms of SOCS in the cervix.

An update on the treatment of gait problems in cerebral palsy.

This article summarizes our experience with cerebral palsy over the past 20 years. The primary and secondary deformities that occur with cerebral palsy are described. Following this, there is a brief overview of the nature and role of gait analysis in the treatment of gait problems in cerebral palsy. The concept of lever-arm dysfunction is introduced. Our current treatment algorithm is then presented along with a brief discussion of our current treatment program, which is illustrated by a case example. Finally, a brief study of a group of patients with spastic diplegia or quadriplegia is presented to illustrate our current method of evaluating treatment outcomes and the need for team management in the treatment of this complex condition.

Viral interleukin 6 stimulates human peripheral blood B cells that are unresponsive to human interleukin 6.

Cellular responsiveness to human interleukin 6 (hIL6) requires the expression of two receptor molecules: IL6-specific receptor (CD126'IL6R') and a nonspecific signal-transducing molecule (CD130'gp130'). Regulation of responsiveness to hIL6 is generally controlled by CD126'IL6R' expression. A viral homologue of hIL6 (vIL6) is encoded by human herpesvirus-8 and has biologic activity similar to hIL6 on a number of cell lines. vIL6 differs from hIL6 in its receptor utilization, requiring only CD130'gp130'. Total human B cells isolated from peripheral blood, which are predominantly CD126'IL6R'-negative, as well as sorted CD126'IL6R'-negative B cells, could be stimulated by recombinant vIL6, but not by hIL6, as indicated by induction of IL6-like signaling (STAT3 phosphorylation). This suggests that the ability of vIL6 to stimulate B cells expressing little or no CD126'IL6R' allows it to act on a larger pool of target B cells, compared to human IL6.

Effects of human papillomavirus-associated cells on human immunodeficiency virus gene expression.

OBJECTIVE: To examine the effects of soluble factors secreted by human papillomavirus (HPV)-associated cells on human immunodeficiency virus (HIV) expression. METHODS: Supernatants collected from cultured cervical biopsies and cervical cancer cell lines, and HPV-immortalized and normal keratinocytes were tested for the ability to induce HIV p24 production in two cell lines that contained latent HIV (the U1 monocytic line and the ACH-2 T cell line). Levels of HIV p24 were measured by enzyme-linked immunosorbent assay (ELISA). Culture supernatants were also assayed for the inflammatory cytokines interleukin 6, tumor necrosis factor, and interleukin 1 beta by ELISA. RESULTS: Supernatants from all epithelial cells tested upregulated HIV p24 expression in the U1 line but not in the ACH-2 cells. Only differentiated normal keratinocytes induced p24 production by ACH-2 cells. Neutralization of the cytokines, particularly interleukin 6, partially reduced the level of HIV-inducing activity in the culture supernatants. Additionally, cervical biopsies from HIV-infected women cultured in vitro also were able to induce HIV in U1 cells but not ACH-2 cells. CONCLUSIONS: Our results suggest that HPV infection of the cervix might influence HIV pathogenesis by inducing the production of immune and inflammatory factors that enhance HIV expression.

An index for quantifying deviations from normal gait.

A method is derived to calculate the amount by which a subject's gait deviates from an average normal profile, and to represent this deviation as a single number. The method uses principal component analysis to derive a set of 16 independent variables from 16 selected gait variables. The sum of the square of these 16 independent variables is interpreted as the deviation of the subject's gait from normal. Statistical tests of the method's validity and an initial demonstration of its clinical utility are included. It is found that using this index, increasing clinical involvement corresponds to increasing index score.

Human herpesvirus 8-encoded interleukin 6 activates HIV-1 in the U1 monocytic cell line.

OBJECTIVE: Human herpesvirus 8 (HHV-8) encodes a viral interleukin 6 (vIL-6) which is structurally and functionally similar to human interleukin 6 (hIL-6). Since hIL-6 has been shown to upregulate the expression of HIV-1, the objectives of this study were to examine the ability of vIL-6 to upregulate HIV-1, and to determine the interactions of this virokine (viral cytokine) with the components of the interleukin 6 (IL-6) receptor complex. DESIGN AND METHODS: Recombinant HHV-8 vIL-6 (rvIL-6) was assayed for bioactivity in the IL-6-dependent cell line MH60.BSF2. HIV-1 p24 production by the U1 monocytic and ACH-2 T-cell lines, which are chronically infected with HIV-1, was used to assess the ability of vIL-6 to affect HIV-1 expression. hIL-6 and vIL-6 receptor utilization was determined by quantifying HIV-1 p24 production after neutralization of components of the IL-6 receptor complex, CD126'IL-6R' and CD130'gp130', on U1 cells with blocking antibodies. RESULTS: HHV-8 rvIL-6 was seen to have IL-6-like bioactivity in MH60.BSF2 cells, and readily upregulated HIV-1 p24 production in U1 monocytic cells, but not in ACH-2 T cells. The vIL-6 appeared to utilize the IL-6-specific component of the IL-6 signaling complex, CD126'IL-6R', in U1 cells. CONCLUSIONS: HHV-8 vIL-6 clearly has the potential to upregulate HIV-1 expression in monocytic cells, and therefore may play a role in AIDS pathogenesis in individuals infected with both viruses.

Lengths of hamstrings and psoas muscles during crouch gait: effects of femoral anteversion.

Recent studies of muscle lengths measured by means of gait analysis data and musculoskeletal models have suggested that in many cases of crouch gait in patients with cerebral palsy, the hamstrings are of normal length and the psoas muscles are short. In these studies, however muscle lengths were calculated by applying kinematic data from a child's joint to a normal adult model. Children with cerebral palsy and other disorders generally do not have normal bone architecture but instead have muscle attachment points and muscle paths altered by osseous deformities. In this study, we explored the consequences of using normal adult musculoskeletal models to calculate hamstring and psoas lengths for children with cerebral palsy. Specifically, for a group of subjects with cerebral palsy who walk with a crouch gait, we investigated the changes in muscle lengths that arise when a patient-specific representation of clinically measured femoral anteversion was added to a model of normal musculoskeletal geometry. The calculation of psoas muscle length was found to be very sensitive to femoral anteversion whereas the calculation of hamstrings length was found to be relatively insensitive to this osseous deformity.

Differential regulation of IL-6 gene transcription and expression by IL-4 and IL-10 in human monocytic cell lines.

IL-4 and IL-10 inhibit the cytokine production and mRNA expression by monocytes/macrophages. To investigate the molecular mechanism of the inhibitory effect on transcriptional or post-transcriptional regulation of IL-6 gene expression by IL-4 and IL-10, we studied IL-6 production, expression level of IL-6 mRNA, IL-6 promoter activity, transcriptional activity of NF-kappaB and NF-IL-6, and IL-6 mRNA stability in human monocytic cell lines, THP-1 and U937, stimulated by PMA and LPS in the absence or the presence of IL-4 or IL-10. Both IL-4 and IL-10 were seen to inhibit IL-6 production and the expression of IL-6 mRNA in both monocytic cell lines studied. In chloramphenicol acetyltransferase assays, utilizing the transient transfection of a chloramphenicol acetyltransferase reporter plasmid containing the IL-6 gene promoter, IL-4, but not IL-10, suppressed the transcriptional activity of the IL-6 gene promoter stimulated by PMA and LPS. Electrophoretic mobility shift assays showed that IL-4, but not IL-10, inhibited nuclear NF-kappaB activity, and that IL-4 and IL-10 did not affect NF-IL-6 activity. On the other hand, IL-10 enhanced the degradation of IL-6 mRNA in a mRNA stability assay. These results suggest that IL-4 may inhibit the transcription of the IL-6 gene by affecting NF-kappaB binding activity, while IL-10 may inhibit the IL-6 mRNA levels post-transcriptionally, without suppressing promoter activity. Therefore, we conclude that IL-4 and IL-10 inhibit IL-6 production by different mechanisms in human monocytic cell lines.

Kinematic and kinetic evaluation of the ankle after lengthening of the gastrocnemius fascia in children with cerebral palsy.

The effect of surgical lengthening of the gastrocnemius fascia on ankle joint kinematics and kinetics during gait in patients with cerebral palsy (CP) was evaluated. Twenty independent ambulators (24 sides) were included in this retrospective study. The evaluation included clinical examination, calculation of joint kinematics and kinetics, and collection of surface electromyography (EMG) during gait. Postoperative improvements were noted in static heelcord range of motion (ROM), with an associated increase in dorsiflexion in stance and swing. Kinetic analysis showed a decrease in the abnormal energy generated around the ankle in midstance and a statistically significant increase in the energy generated in late stance for push-off.

Rectus femoris surgery in children with cerebral palsy. Part I: The effect of rectus femoris transfer location on knee motion.

Rectus femoris transfer was performed in 78 children (105 sides) with cerebral palsy (CP) at the same time as other surgical procedures as appropriate. The transfer was either medial to the sartorius (62 sides), semitendinosus (19 sides), or the gracilis (14 sides) muscles, or laterally to the iliotibial band (10 sides). Gait analysis performed before and 1 year after operation demonstrated increased knee range of motion (ROM) with increased extension at initial contact and in midstance and maintained knee flexion in swing. There were no statistically significant differences between the four transfer sites in the effect on those variables. Therefore, the choice of rectus femoris transfer site can be dictated by surgical preference or by the nature of other simultaneous procedures. There was no consistent change in transverse plane motion of the hip or foot progression angles between the two gait analyses, suggesting that rectus femoris transfer does not affect gait abnormalities observed in the transverse plane.

Rectus femoris surgery in children with cerebral palsy. Part II: A comparison between the effect of transfer and release of the distal rectus femoris on knee motion.

Rectus femoris muscle (RF) surgery was performed in 98 children (136 sides) with cerebral palsy (CP). RF transfer was performed in 105 lower limbs, and distal RF release was performed in 31. Eleven (20 sides) similarly affected children had no RF procedure and are included for comparison. Gait analysis was performed just before and approximately 1 year after surgery. All children underwent other orthopaedic surgery at the time of the RF procedure. When preoperative knee range of motion (ROM) was > 80% of normal, there were no significant changes in knee motion in either the RF transfer or distal release groups. In patients with < 80% of normal knee ROM preoperatively, RF transfer was followed by maintained knee flexion in swing; patients who underwent distal RF release or no RF procedure showed a decrease (10 degrees and 6 degrees, respectively) in knee flexion postoperatively. These results suggest that the RF should be transferred and not released when knee ROM is < 80%.

Effects of selective dorsal rhizotomy on gait in children with cerebral palsy.

We wished to examine the effects of selective dorsal rhizotomy (SDR) on the gait patterns of children with cerebral palsy (CP). Nineteen ambulatory children underwent preoperative and 1-year postoperative gait analyses that included clinical assessment of joint range of motion (ROM) and muscle tone, three-dimensional motion analysis, and dynamic electromyography (EMG). The children were divided into two groups: independent ambulators (n = 11) and dependent ambulators (requiring a walking aid) (n = 8). Improvements in joint passive ROM and a reduction in lower extremity spasticity were noted. In both groups, positive pre- to postoperative improvements were noted in sagittal plane hip, knee, and ankle motion and there was a greater incidence of a plantar-grade foot position in stance. There was no change in the coronal plane motion of the pelvis and hip. A mean increase in anterior pelvic tilt for the independent ambulators was the only major negative change observed in this study. We conclude that SDR improves specific gait parameters in children with CP.

Gait analysis. An essential tool in the treatment of cerebral palsy.

Gait analysis has radically changed the treatment of cerebral palsy. Preoperatively, it allows critical assessment of the specific pathologies of the patient. Postoperatively, it provides an accurate assessment of outcome. This assessment of outcome has in turn allowed the accurate critique of surgeries and has made it possible to discard treatments that are not useful or are perhaps even injurious. As a result of this continual reassessment of surgical techniques, several principles and insights regarding the treatment of cerebral palsy have been learned. These include (1) the importance of reestablishing normal gait prerequisites, (2) the methods of reducing the energy expenditure of the pathologic gait, (3) the importance of skeletal structures in providing the lever arm by which muscles produce moments around joints, (4) the role and importance of two joint muscles, and (5) the importance of separating abnormalities, which are emanating from the neurologic lesion, from secondary ("coping") responses. Through gait analysis, it has become apparent that diplegia and hemiplegia are noninclusive terms, each of which contain a variety of homogeneous patterns of gait. Eventually these patterns may be separated and identified and optimal treatment protocols for each pattern type developed.

[Treatment of deformities of the locomotor system in hemiplegia]. / Die Behandlung der Deformitäten am Bewegungsapparat bei Hemiplegie.

In spastic hemiplegia mainly one side of the body is affected. In both the upper and the lower extremity the distal parts (hand and foot) are more severely involved than the proximal region. In cases of minor involvement the goal of treatment in the upper extremity is to achieve functional improvement by means of splinting and surgery. In cases of severe alterations cosmetic improvement without much functional gain is all that can be expected. Gait analysis has demonstrated that there are four basic patterns that can be related to the severity of involvement. In type I muscle imbalance exists without a contracture. In type II there is contracture of the muscles of the posterior compartment of the calf. In type III, in addition to the changes around the ankle joint, contractures around the knee are present, and in type IV also hip problems. Functional improvement can be achieved by means of splinting and surgery in all types. Basic principles of treatment have developed as a result of the application of gait analysis and dynamic electromyography. Specific examples of such treatment principles have recently been presented by Gage.

Clinical determination of femoral anteversion. A comparison with established techniques.

We evaluated femoral anteversion preoperatively in fifty-nine patients (ninety-one hips), using a clinical method that we developed, Magilligan radiographs, and computed tomographic scans. These measurements were then compared with values for anteversion that were obtained intraoperatively. To determine femoral anteversion clinically, the patient was placed in the prone position and the maximum lateral trochanteric prominence was related to the degree of internal rotation of the hip. Compared with computed tomographic scanning and Magilligan radiographic determination, the clinically determined anteversion correlated most closely (to within 4 degrees) with the amount measured at the time of the operation. The clinical method was found to be superior to radiographic techniques for determination of the degree of femoral anteversion in children who have not had a previous operation about the hip.

Human papillomavirus type 16 immortalized cervical keratinocytes contain transcripts encoding E6, E7, and E2 initiated at the P97 promoter and express high levels of E7.

Human cervical keratinocytes represent the specific host for the genital human papillomaviruses (HPV). Transfection of these cells with the DNA of a number of the oncogenic HPVs including type 16 was recently shown to result in their immortalization but not in malignant transformation. In this report we show that viral transcripts for E6 and E7 in these cells were as abundant as in cancer derived cell lines. However, in contrast to cancer derived cell lines, immortalized cervical keratinocytes contained RNA with the potential to encode a full-length E2 protein. In addition, the levels of the E7 oncoprotein were at least as high as in cancer derived cell lines, suggesting that E2 interruption, observed in cancer derived cell lines, is not causally related to the high level of E7 expression and, therefore, deregulation of the P97 promoter may not be a prerequisite for HPV-16 associated cancer development. Furthermore, we show that E6, E7, and E2 encoding transcripts all originate from the viral promoter, P97. Unlike in cancer derived cell lines, all transcripts terminated at the early poly(A) site.