Phase-contrast 3D tomography of HeLa cells grown in PLLA polymer electrospun scaffolds using synchrotron X-rays.

Advanced imaging is useful for understanding the three-dimensional (3D) growth of cells. X-ray tomography serves as a powerful noninvasive, nondestructive technique that can fulfill these purposes by providing information about cell growth within 3D platforms. There are a limited number of studies taking advantage of synchrotron X-rays, which provides a large field of view and suitable resolution to image cells within specific biomaterials. In this study, X-ray synchrotron radiation microtomography at Diamond Light Source and advanced image processing were used to investigate cellular infiltration of HeLa cells within poly L-lactide (PLLA) scaffolds. This study demonstrates that synchrotron X-rays using phase contrast is a useful method to understand the 3D growth of cells in PLLA electrospun scaffolds. Two different fiber diameter (2 and 4 µm) scaffolds with different pore sizes, grown over 2, 5 and 8 days in vitro, were examined for infiltration and cell connectivity. After performing visualization by segmentation of the cells from the fibers, the results clearly show deeper cell growth and higher cellular interconnectivity in the 4 µm fiber diameter scaffold. This indicates the potential for using such 3D technology to study cell-scaffold interactions for future medical use.

A small molecule ligand for the novel pain target, GPR171, produces minimal reward in mice.

ProSAAS is one of the most abundant proteins in the brain and is processed into several smaller peptides. One of which, BigLEN, is an endogenous ligand for the G protein-coupled receptor, GPR171. Recent work in rodent models has shown that a small-molecule ligand for GPR171, MS15203, increases morphine antinociception and is effective in lessening chronic pain. While these studies provide evidence for GPR171 as a possible pain target, its abuse liability has not yet been assessed and was evaluated in the current study. We first mapped the distribution of GPR171 and ProSAAS throughout the reward circuit of the brain using immunohistochemistry and showed that GPR171 and ProSAAS are localized in the hippocampus, basolateral amygdala, nucleus accumbens, prefrontal cortex. In the major dopaminergic structure, the ventral tegmental area (VTA), GPR171 appeared to be primarily localized in dopamine neurons while ProSAAS is outside of dopamine neurons. Next, MS15203 was administered to mice with or without morphine, and VTA slices were stained for the immediate early gene c-Fos as a marker of neuronal activation. Quantification of c-Fos-positive cells revealed no statistical difference between MS15203 and saline, suggesting that MS15203 does not increase VTA activation and dopamine release. The results of a conditioned place preference experiment showed that treatment with MS15203 produced no place preference indicating a lack of reward-related behavior. Taken together this data provides evidence that the novel pain therapeutic, MS15203, has minimal reward liability. Therefore, GPR171 deserves further exploration as a pain target. SIGNIFICANCE STATEMENT: MS15203, a drug that activates the receptor GPR171, was previously shown to increase morphine analgesia. The authors use in vivo and histological techniques to show that it fails to activate the rodent reward circuitry, providing support for the continued exploration of MS15203 as a novel pain drug, and GPR171 a novel pain target.

Instantaneous 4D micro-particle image velocimetry (µPIV) via multifocal microscopy (MUM).

Multifocal microscopy (MUM), a technique to capture multiple fields of view (FOVs) from distinct axial planes simultaneously and on one camera, was used to perform micro-particle image velocimetry (µPIV) to reconstruct velocity and shear stress fields imposed by a liquid flowing around a cell. A diffraction based multifocal relay was used to capture images from three different planes with 630 nm axial spacing from which the axial positions of the flow-tracing particles were calculated using the image sharpness metric. It was shown that MUM can achieve an accuracy on the calculated velocity of around (0.52 ± 0.19) µm/s. Using fixed cells, MUM imaged the flow perturbations at sub-cellular level, which showed characteristics similar to those observed in the literature. Using live cells as an exemplar, MUM observed the effect of changing cell morphology on the local flow during perfusion. Compared to standard confocal laser scanning microscope, MUM offers a clear advantage in acquisition speed for µPIV (over 300 times faster). This is an important characteristic for rapidly evolving biological systems where there is the necessity to monitor in real time entire volumes to correlate the sample responses to the external forces.

Video-assisted treatment of thoracolumbar junction fractures using a specific distractor for reduction: prospective study of 50 cases.

Posterior instrumentation allows good osteosynthesis for thoracolumbar junction fractures. However, in approximately 20% of cases, anterior bone defects may persist, leading to pseudoarthrosis and loss of reduction. Anterior approaches can circumvent this drawback, but they are considered aggressive with a high rate of complications. The advent of the endoscopic mini-invasive techniques has led to a reduction in the number of complications while maintaining the same consolidation rate. Nevertheless, poor restitution of anatomic curves is a reproach for this technique. This prospective study reports clinical and radiological result of 50 patients (19 women and 31 men) operated between April 2000 and March 2006 with a video-assisted anterior approach using an endodistractor for reduction and consequent insertion of the implant. There were A3 (n = 44), A2 (n = 2), A1 (n = 3) and C1 (n = 1) fractures (Magerl classification). The specific system for fracture reduction was used in the last 39 cases of this series. A Pyramesh cage (Medtronic, Memphis, USA) was used in 15 patients, a peek cage (Creaspine, Bordeaux, France) in 30 patients and a tricortical graft in 5 patients. Standard X-rays and CT scan were performed pre-operatively. Kyphosis, and angulations were measured pre-, per- and post-operatively. Mean immediate postoperative gain in localized kyphosis was 12.18 degrees and mean gain at last follow-up was 11.71 degrees. Mean immediate postoperative gain in RA was 13.24 degrees and remained identical at last follow-up. Five patients had a transient pulmonary atelectasia and there was one pulmonary infection. No neurological complication occurred. Fracture reduction is comparable to the best thoracotomy series while limiting approach-related complications.

Posterolateral fusion versus Dynesys dynamic stabilization: Retrospective study at a minimum 5.5years' follow-up.

INTRODUCTION: Lumbar fusion is one of the most widespread techniques to treat degenerative lumbar pathology. To prevent complications such as non-union or adjacent segment degeneration, dynamic stabilization techniques were developed, but with controversial results. The aim of the present study was to compare long-term radiologic and clinical results between fusion and dynamic stabilization. MATERIAL AND METHODS: A single-center retrospective study included patients with recurrent lumbar discal hernia or lumbar canal stenosis managed by posterolateral fusion or by dynamic stabilization associated to neurologic release. Patients were seen in follow-up for radiological and clinical assessment: visual analog pain scale (VAS), Oswestry Disability Index (ODI), Short Form-12 (SF-12), adjacent segment disease (ASD), and intervertebral range of motion (ROM). RESULTS: Fifty-eight patients were included: 25 in the fusion group (FG), and 33 in the Dynesys® group (DG). VAS scores were significantly lower in DG than FG. ODI was 14.6±2.8 in DG, versus 19.4±3.3 in FG (P=0.0001). SF-12 physical subscore was significantly higher in DG. ROM was 4.1±2° in DG, vs. 0.7±0.5° in FG (P=0.001). Radiologic ASD was significantly greater in FG than DG (36% vs. 12.1%; P=0.012), without difference in clinical expression (DG, 1 case; FG, 2 cases). CONCLUSION: Dynamic stabilization provided clinical and radiological results comparable to those of posterolateral fusion in these indications (although level L5-S1 was not studied). LEVEL OF EVIDENCE: IV.

Visualising the 3D microstructure of stained and native intervertebral discs using X-ray microtomography.

Intervertebral disc degeneration (IVDD) is linked to low back pain. Microstructural changes during degeneration have previously been imaged using 2D sectioning techniques and 3D methods which are limited to small specimens and prone to inducing artefacts from sample preparation. This study explores micro computed X-ray tomography (microCT) methods with the aim of resolving IVD 3D microstructure whilst minimising sample preparation artefacts. Low X-ray absorption contrast in non-mineralised tissue can be enhanced using staining and phase contrast techniques. A step-wise approach, including comparing three stains, was used to develop microCT for bovine tail IVD using laboratory and synchrotron sources. Staining successfully contrasted collagenous structures; however not all regions were stained and the procedure induced macroscopic structural changes. Phase contrast microCT of chemically fixed yet unstained samples resolved the nucleus pulposus, annulus fibrosus and constituent lamellae, and finer structures including collagen bundles and cross-bridges. Using the same imaging methods native tissue scans were of slightly lower contrast but free from sample processing artefacts. In the future these methods may be used to characterise structural remodelling in soft (non-calcified) tissues and to conduct in situ studies of native loaded tissues and constructs to characterise their 3D mechanical properties.

[Anatomic variations of the ilio-caval junction encountered in disk prosthesis implantation]. / Incidence de la situation anatomique de la bifurcation ilio-cave lors de la mise en place d'une prothèse discale.

PURPOSE OF THE STUDY: The retroperitoneal mini-invasive anterior approach to the lumbar spine is widely used for disc excision and insertion of a prosthetic disc. A large operating window is needed. Venous wounds constitute one of the most serious complications. We analyzed the angle and position of the iliocaval on the preoperative angio-MRI in search for correlations with the intraoperative findings. Our aim was to determine whether the preoperative angio-MRI gives indications concerning the operative difficulty and the best strategy for prosthesis fitting. MATERIAL AND METHOD: This prospective study included 35 consecutive patients who underwent surgery for implantation of a Maverick disc prosthesis. Average patient age was 46.7 years. The indication for surgery was isolated degenerative discopathy with invalidating chronic low back pain without alteration of the muscle masse or facet joints. A preoperative angio-MRI was performed with T1 spin-echo sequences for the coronal slices and T2 axial slices passing through the upper extremity of the L5-S1 disc. We searched for correlations between the MRI and operative findings. Elements studied were those used in the Capellades classification: height of the iliocaval junction (high, very high, low, very low), position of the common iliac vein (lateral, intermediate, medial), angle formed by the two common iliac veins. RESULTS: The lateral position was the most frequent (31.5%). The average junction angle was 65 degrees). The only position where it was not possible to "pass" under the iliocaval junction was for a very low medial localization with a narrow angle (45 degrees). DISCUSSION: Our series included a homogenous group comparable with other series in the literature. The junction angle for very low medial localizations is of considerable importance because it is impossible to fit the implant in the L5-S1 under the iliocaval junction if the angle is over 60 degrees. CONCLUSION: The position of the iliocaval junction, and particularly its angle, are of considerable importance for insertion of an L5-S1 disc prosthesis. The preoperative angio-MRI provides information on the potential difficulty of the insertion. In addition, with this preoperative information, the patient can be warned that it may not be possible to insert the implant so that a therapeutic alternative may be proposed.

Endochondral Growth Defect and Deployment of Transient Chondrocyte Behaviors Underlie Osteoarthritis Onset in a Natural Murine Model.

OBJECTIVE: To explore whether aberrant transient chondrocyte behaviors occur in the joints of STR/Ort mice (which spontaneously develop osteoarthritis [OA]) and whether they are attributable to an endochondral growth defect. METHODS: Knee joints from STR/Ort mice with advanced OA and age-matched CBA (control) mice were examined by Affymetrix microarray profiling, multiplex polymerase chain reaction (PCR) analysis, and immunohistochemical labeling of endochondral markers, including sclerostin and MEPE. The endochondral phenotype of STR/Ort mice was analyzed by histologic examination, micro-computed tomography, and ex vivo organ culture. A novel protocol for quantifying bony bridges across the murine epiphysis (growth plate fusion) using synchrotron x-ray computed microtomography was developed and applied. RESULTS: Meta-analysis of transcription profiles showed significant elevation in functions linked with endochondral ossification in STR/Ort mice (compared to CBA mice; P < 0.05). Consistent with this, immunolabeling revealed increased matrix metalloproteinase 13 (MMP-13) and type X collagen expression in STR/Ort mouse joints, and multiplex quantitative reverse transcriptase-PCR showed differential expression of known mineralization regulators, suggesting an inherent chondrocyte defect. Support for the notion of an endochondral defect included accelerated growth, increased zone of growth plate proliferative chondrocytes (P < 0.05), and widespread type X collagen/MMP-13 labeling beyond the expected hypertrophic zone distribution. OA development involved concomitant focal suppression of sclerostin/MEPE in STR/Ort mice. Our novel synchrotron radiation microtomography method showed increased numbers (P < 0.001) and mean areal growth plate bridge densities (P < 0.01) in young and aged STR/Ort mice compared to age-matched CBA mice. CONCLUSION: Taken together, our data support the notion of an inherent endochondral defect that is linked to growth dynamics and subject to regulation by the MEPE/sclerostin axis and may represent an underlying mechanism of pathologic ossification in OA.

[Video-assisted mini-thoracotomy for surgical treatment of thoracolumbar junction fractures]. / Traitement des fractures de la charnière thoraco-lombaire par thoracotomie mini-invasive vidéo assistée: a propos de 20 cas.

PURPOSE OF THE STUDY: There are several solutions for the treatment of fractures of the thoracolumbar junction (classical anterior, posterior, combined approaches). The advent of video-assisted minimally invasive techniques has helped minimize complications. The aim of this work was to analyze the clinical and radiological outcome in a prospective series of 20 patients who underwent video-assisted mini-thoracotomy for the treatment of thoracolumbar junction fractures. MATERIAL AND METHODS: This prospective study included 20 patients, mean age 43.3 years, with at least one year follow-up. All patients presented a normal neurological examination (Frankel E). Fractures were classified Magerl A (n = 19) and B (n = 1). Corporectomy/correction (aided with an endodistractor in the last nine cases) was performed with grafting and osteosynthesis. Perioperative and postoperative complications were noted. Focal kyphosis and regional angulation were noted before and after the operation and at last follow-up using the Stagnara system. RESULTS: The arthrodesis had fused at last follow-up in all patients. There were three cases of alelectasia which regressed in two months. Overall outcome showed satisfactory angular correction which was maintained at last follow-up. The gain in focal kyphosis was 13.71 degrees on average in the immediate postoperative period and persisted at last follow-up (13.31degrees). The gain in corrected regional angulation was 14 degrees in the immediate postoperative period and 14 degrees at last follow-up. The gain in focal kyphosis and regional angulation was two-fold greater with the endodistractor than with external maneuvers. DISCUSSION: The complication rate was very low. Corrections obtained were comparable with those reported in the literature and persisted over time. CONCLUSION: Bearing in mind the learning curve, there are fewer complications with the video-assisted minimally invasive approach than with classical thoracotomy or anterior surgery. This is true for perioperative and early and late postoperative complications. The correction achieved is satisfactory and lasting. A specific ancillary is essential for the reduction and for fitting the anterior graft without lost of correction. This technique combines the best results achieved with thoracotomy (Onimus) with a limited rate of complications.

[High tibial open-wedge osteotomy using a tricalcium phosphate substitute: 70 cases with 18 months mean follow-up]. / Valgisation tibiale par ouverture médiale utilisant un coin de céramique de phosphate tricalcique: á propos de 70 cas revus avec un recul moyen de 18 mois.

PURPOSE OF THE STUDY: We report our experience with a B tricalcium phosphate ceramic to fill open-wedge tibial osteotomy gaps. MATERIAL AND METHODS: Seventy high tibial open-wedge osteotomies were performed in 70 patients (47 men and 23 women). Ostotomy was performed for osteoarthritis in 56 knees, Ahlback classification stage I (n = 18), stage II (n = 32), stage III (n = 6) and for congenital varus in 14. Mean patient age was 45.2 years (16-69). We used a wedge-shaped piece of ceramic with an appropriate angle, which was associated with granulated material to complete the gap filling for the last 20 cases. One or two staples were used for fixation. Clinical and radiological outcome was assessed at 6 weeks, 3 months, 6 months, and at last follow-up. Mean follow-up was 18 months. RESULTS: There were no biological or clinical complications related to biomaterial intolerance. Tolerance to the osteosynthesis material was mediocre since we observed one infection and 12 cases of pain related to the staples which required material removal in 8 patients. Bone healing was achieved in 98.5%. Final correction was between 3 degrees and 6 degrees valgus in 80.5%. There was a loss of correction angle between the postoperative film and the bone healing film. Factors related to sustained correction were: non-fractured wedge, intact lateral tibial cortical, osteosynthesis with two staples. Osteointegration was good and rapid in 96%. We found two complete lucent lines at last follow-up but with variable resorption depending on the shape of the bone substitute. CONCLUSION: Use of a ceramic wedge to fill high tibial medial open-wedge osteotomy gap is a reliable reproducible technique providing correction without formation of a malalignment callus. Bone healing is achieved in 98.5% of the cases at about three months. Osteointegration is good in 96%. Resorption is complete and rapid when the substitute is implanted in granular form in a cancellous zone and is partial and slow when implanted as a massive wedge.

[Pertrochanteric fractures: a randomized prospective study comparing dynamic screw plate and intramedullary fixation]. / Comparaison vis-plaque dynamique et ostéosynthèse intra-médullaire antérograde dans les fractures pertrochantériennes: une étude prospective randomisée.

PURPOSE OF THE STUDY: The objective of this study was to compare the dynamic hip screw (Synthes) and intramedullary fixation (Targon PF, Aesculap) for the treatment of pertrochanteric fractures in terms of stability, complications and cost effectiveness. MATERIAL AND METHODS: This prospective randomized study included 60 patients hospitalized in an emergency setting for pertrochanteric fractures between December 2003 and June 2004. All surgeons in our unit participated in the study. Two fixation systems were used: the Targon proximal femoral nail (Aesculap) and the screw-plate dynamic hip screw (Synthes). We noted: patient status (ASA classification), operative data (type of implant, duration), postoperative data (blood loss, radiographic findings, early complications), and outcome (Harris score, time to walking, mortality). All patients were assessed three months after surgery. The series included 60 patients, 14 men (23%) and 46 women (77%). The Targon PF nail was used for 34 patients and the DHS for 26. In the intramedullary fixation group mean age was 81 years (SD = 12.8, range 23-96); it was 82 years (SD 9.8, range 47-97) in the screw-plate group. The AO classification was: intramedullary fixation 31A1 (n = 11), 31A2 (n = 20), 31A3 (n = 3); screw-plate 31A1 (n = 14), 31A2 (n = 11), 31A3 (n = 1). Thirty-one fractures were stable, 29 unstable. Unstable fractures were treated with the Targon PF nail (n = 18) and the DHS screw-plate (n = 11). Stable fractures were treated with the Targon PF nail (n = 15) and the DHS screw-plate (n = 15). RESULTS: Mean operative time was 35 minutes for intramedullary nailing and 42 mintues for screw-plate fixation. Mean blood loss was 410 ml for intramedullary nailing and 325 ml for screw-plate fixation (p = 0.07). Mean hospital stay was the same (11 days) in both groups. At three months, mechanical complications involved migration of the cervical screw outside the femoral head for three Targon PF fixations and for two DHS fixations. Screw migration was favored by fracture instability and presence of osteoporosis. There were two deaths in the intramedullary nailing group and one in the screw-plate group. Time to walking was 20 days on average in the intramedullary nailing group and 25 days in the screw-plate group. The mean Harris hip score was 60 in the intramedullary group and 59 in the screw-plate group. DISCUSSION AND CONCLUSION: Data in the literature report an advantage for intramedullary nailing, particularly a mechanical advantage, for the treatment of pertrochanteric fractures. Our findings show that good results are obtained with the screw-plate fixation using the DHS with less blood loss and at a lower cost.

Macrophage migration inhibitory factor promotes eosinophil accumulation and tissue remodeling in eosinophilic esophagitis.

Macrophage migration inhibitory factor (MIF) is involved in eosinophil biology and in type 2 inflammation, contributing to allergic and helminthic diseases. We hypothesized that MIF participates in the pathogenesis of eosinophilic esophagitis (EoE), an allergic condition characterized by esophageal eosinophilic inflammation. MIF is highly expressed in esophageal mucosa of patients with EoE, compared with gastro-esophageal reflux disease and control patients, where it co-localizes predominantly with eosinophils. In vitro, recombinant MIF promotes human eosinophil chemotaxis, while MIF antagonist and CXCR4 antagonist, AMD3100, revert this effect. In a model of EoE induced by ovalbumin, Mif-deficient mice have reduced inflammation and collagen deposition compared with wild-type (WT) mice. Importantly, treatment of WT mice with anti-MIF or with AMD3100 during the challenge phase prevents accumulation of eosinophils and tissue remodeling. Conversely, recombinant MIF promoted tissue eosinophil inflammation in allergic mice. Together, these results implicate MIF in the pathogenesis of esophageal inflammation and suggest that targeting MIF might represent a novel therapy for EoE.

Jejunal biopsy in HIV-infected patients.

Forty-nine HIV-infected patients were submitted to peroral jejunal biopsy in order to evaluate the presence of microorganisms and the histomorphometric aspects of the enteric mucosa with subsequent correlation of these findings to the appropriate clinical stage of the disease. Thirty-seven patients fulfilled the CDC criteria for AIDS, of whom 23 presented with diarrhea. Of the 12 patients who had not yet been given an AIDS diagnosis. 3 had persistent generalized lymphadenopathy and 9 were asymptomatic carriers. Flat mucosa was observed in two patients (8.7%) with diarrhea and coccidea. Subtotal villous atrophy and severe lamina propria (LP) mononuclear infiltrate (13%) were found only in patients with diarrhea. Moderate to severe histologic changes were more frequently observed in this group, not always related to the presence of microorganisms. Crypt hyperregeneration was a constant finding. Intraepithelial lymphocyte (IEL) count was decreased in patients with diarrhea. Specific infectious agents were unexpectedly rare for the tropical developing country population studied. The organism most commonly associated with diarrhea was Cryptosporidium sp. (21.7%). The etiology of diarrhea in a significant number of patients remains unclear.

Evolving features of lung adenocarcinoma in Rio de Janeiro, Brazil.

Formerly considered rare, adenocarcinoma has become the commonest form of primary lung cancer in developed countries. Its clinical presentation has changed, with central tumors becoming more frequent. We reviewed all biopsies with a diagnosis of primary lung cancer obtained from October 1988 to April 1997 in a tertiary care hospital in Rio de Janeiro. Medical records from adenocarcinomas were analysed. Adenocarcinoma was the commonest form of lung cancer (168/409, 41%). Central tumors were observed in 43% according to radiological criteria and 47% according to bronchoscopic criteria. The frequency and clinical presentation of adenocarcinoma have evolved lately in Rio de Janeiro.

p53 as tumor marker for the early detection of reincidence in lung cancer.

To examine the p53 expression in cancer development, we studied the case of a 59-year-old male suffering from squamous cell lung cancer. He was submitted to surgery for resection of the tumoral mass. Through immunoblot and immunohistochemical analysis, the tumor fragment was shown to bear p53 and hsp70 accumulation not detected in the normal lung tissue sample. Six months later, immunohistochemical analysis of a biopsy sample taken from the previous tumor site showed p53 accumulation. Considering the high specificity for p53 detection (100%), this result suggests probable tumor reincidence, not detected by the ordinary H&E staining.

Expression of heat shock protein 70 and p53 in human lung cancer.

Bronchial biopsies of 21 patients with lung cancer were analyzed by Western blot for stress protein HSP70 and p53 proteins. Squamous carcinoma was the most common type found. The p53 protein was detectable in 14 cases. The HSP70 was detectable in 17 and overexpressed in 9 patients. Eleven patients showed positivity for both protein expressions, however no statistical significance was found (Kappa's test, p>0.05). Specific associations were not observed for HSP70 overexpression and p53 detection that could be related to clinical finds or tabagism. Our results indicate that the stress protein HSP70 is detectable and may be involved in the tumor development.

Phenotypic analysis of intestinal non-inflamed mucosa in Crohn's disease: evidence of mononuclear cell depletion in lamina propria.

OBJECTIVE: The objective of this study was to examine mononuclear cell subpopulations and evidence of cellular activation in unaffected jejunal mucosa in Crohn's disease. DESIGN: A cross-sectional study was performed in patients with Crohn's disease from the ambulatory unity of the University Hospital, UFRJ. METHODS: Mucosal samples from 20 patients with Crohn's colitis or ileitis were obtained by peroral jejunal biopsy. Patients with jejunal involvement or pregnant women were excluded from the study. Specimens were analysed histologically and by indirect immunoperoxidase method using anti-monoclonal antibodies to CD2, CD4, CD8, CD25, CD45RO, RFDR1, RFD1 and RFD7 by two 'blind' observers. Seven patients with non-inflammatory bowel disorders and two healthy volunteers were studied as controls. RESULTS: Lamina propria CD2-positive (CD2+) cells were reduced in Crohn's disease (P < 0.004) whether clinically active (P < 0.02) or clinically inactive (P < 0.008). CD4+ and CD8+ cells were also reduced in Crohn's disease (P < 0.003), whereas the CD4:CD8 ratio did not differ from that in controls. CD25+, CD45RO+ and HLA-DR+ cells were not significantly increased in patients with Crohn's disease. RFD7+ cells were decreased in Crohn's disease (P < 0.02), whereas RFD1+ cells were not significantly different from the control group. CONCLUSION: No evidence of cellular activation was found in the unaffected mucosa of Crohn's disease. The reduction in T-cell and macrophage-like cell numbers may result from cell migration to inflamed areas. It is also possible that this finding represents a primary defect which may have a role in the pathogenesis of Crohn's disease.

Quantitative and qualitative changes in intestinal immunoglobulin-bearing plasma cells in jejunal mucosa of 52 HIV-infected patients.

1. The purpose of the present study was to evaluate the intestinal secretory immunity of HIV-infected patients. Jejunal biopsies were obtained with a Watson capsule from 52 HIV-infected patients and compared to a control group consisting of 12 healthy subjects, matched for age and sex. IgA- and IgM-containing plasma cells were studied by immunofluorescence by two independent observers who had no knowledge of the history of the slides. 2. The IgA and IgM plasma cell density was significantly decreased in HIV-infected patients when compared to the controls (31/52 for IgA and 42/52 for IgM in the HIV group and normal density in all controls). The depletion of intestinal plasma cells was an early feature of HIV disease and did not correlate with diarrhea, enteropathogens or mucosal atrophy. Although IgA plasma cell depletion has been described in HIV infection, this is the first report of IgM cell density reduction. 3. A peripheral fluorescence pattern was observed in the intestinal cells of HIV-infected patients, contrasting with the normal cytoplasmic fluorescence pattern seen in controls. The cells presenting a rim of peripheral fluorescence in HIV disease might correspond to immature B lymphocytes. 4. The intestinal plasma cell depletion may result in hampered mucosal defense in HIV disease, and is likely to be a direct consequence of T lymphocyte function impairment. 5. The data indicate that the intestinal secretory immune system is altered in AIDS and also in the early stages of HIV disease.

Study of the effects of beta-myrcene on rat fertility and general reproductive performance.

beta-Myrcene (MYR) is a monoterpene found in the oils of a variety of aromatic plants including lemongrass, verbena, hop, bay, and others. MYR and essential oils containing this terpenoid compound are used in cosmetics, household products, and as flavoring food additives. This study was undertaken on investigate the effects of MYR on fertility and general reproductive performance in the rat. MYR (0, 100, 300 and 500 mg/kg) in peanut oil was given by gavage to male Wistar rats (15 per dose group) for 91 days prior to mating and during the mating period, as well as to females (45 per dose group) continuously for 21 days before mating, during mating and pregnancy, and throughout the period of lactation up to postnatal day 21. On day 21 of pregnancy one-third of the females of each group were submitted to cesarean section. Resorption, implantation, as well as dead and live fetuses were counted. All fetuses were examined for external malformations, weighed, and cleared and stained with Alizarin Red S for skeleton evaluation. The remaining dams were allowed to give birth to their offspring. The progeny was examined at birth and subsequently up to postnatal day 21. Mortality, weight gain and physical signs of postnatal development were evaluated. Except for an increase in liver and kidney weights, no other sign of toxicity was noted in male and female rats exposed to MYR. MYR did not affect the mating index (proportion of females impregnated by males) or the pregnancy index (ratio of pregnant to sperm-positive females). No sign of maternal toxicity and no increase in externally visible malformations were observed at any dose level. Only at the highest dose tested (500 mg/kg) did MYR induce an increase in the resorption rate and a higher frequency of fetal skeleton anomalies. No adverse effect of MYR on postnatal weight gain was noted but days of appearance of primary coat, incisor eruption and eye opening were slightly delayed in the exposed offspring. On the basis of the data presented in this paper the no-observed-adverse-effect level (NOAEL) for toxic effects on fertility and general reproductive performance can be set at 300 mg of beta-myrcene/kg body weight by the oral route.

Intra-abdominal tuberculosis in acquired immunodeficiency syndrome. Diagnosis and management.

Tuberculosis (TB) in human immunodeficiency virus (HIV) immunosuppressed patients is characterized by extra-pulmonary disease in as many of 70% of them. If intestinal or lymph node involvement occurs, the differential diagnosis between an acute abdomen and other non surgical conditions may be a challenging problem. The authors analyzed eight double infected patients (TB and acquired immunodeficiency syndrome AIDS), who were admitted to the University Hospital (HUCFF) of the Federal University of Rio de Janeiro. This association should be considered when abdominal pain, anemia, fever, weight loss and abdominal lymph node enlargement are present. Bacteriology of body fluids, abdominal ultrasound (US) and computed tomography scans (CT) combined with guided needle aspiration biopsies, barium examination, colonoscopy and laparoscopy, can not only elucidate the diagnosis but also be helpful in assessing an appropriate management. Thus a systematic evaluation often yields an etiology and a correct therapeutic indication reducing the high mortality rate.