Age and synovitis affect the results of the treatment of knee osteoarthritis with Microfragmented Autologous Fat Tissue.

PURPOSE: This study aims to assess the effectiveness of Microfragmented Autologous Fat Tissue (MFAT) treatment for knee osteoarthritis and to investigate whether patients' pre-treatment clinical condition, such as synovitis, correlates with clinical outcomes, to identify potential predicting factors for the success or failure of the treatment. METHODS: In this prospective Cohort Study Level II multicentric trial, consecutive patients with a diagnosis of early/mild osteoarthritis and failure of previous conservative measures were enrolled to undergo diagnostic arthroscopy and a single MFAT injection. Patients were assessed with repeated scoring systems at baseline, 6 months, and 12 months after surgery. The demographic features, the arthroscopic findings, the immunophenotype of injected tissue and the histologic examination of synovia of failed patients were analyzed. RESULTS: Data from 91 patients showed a significant improvement in Lysholm, WOMAC scores at 1-year follow-up (p < 0.001). A significant decrease in VAS score was observed, while a significant improvement of measured flexion angle was registered at 1 year (p < 0.001). No major complications were reported. Age and synovitis were identified as significant factors influencing the clinical outcome (p < 0.05). Body mass index, previous or concomitant procedures, and specific cartilage defects had no influence. The mean number of injected adipose tissue-derived mesenchymal stem cells seem not to correlate with the clinical outcome. CONCLUSION: MFAT is effective in reducing pain when used with a single dose injection in early/mild OA of the knee, without major complications. Age over 60 and synovitis may be predictive for persistent pain at one year and should be considered before indications.

Primary breast cancer stem-like cells metastasise to bone, switch phenotype and acquire a bone tropism signature.

BACKGROUND: Bone metastases represent a common and severe complication in breast cancer, and the involvement of cancer stem cells (CSCs) in the promotion of bone metastasis is currently under discussion. Here, we used a human-in-mice model to study bone metastasis formation due to primary breast CSCs-like colonisation. METHODS: Primary CD44⁺CD24⁻ breast CSCs-like were transduced by a luciferase-lentiviral vector and injected through subcutaneous and intracardiac (IC) routes in non-obese/severe-combined immunodeficient (NOD/SCID) mice carrying subcutaneous human bone implants. The CSCs-like localisation was monitored by in vivo luciferase imaging. Bone metastatic CSCs-like were analysed through immunohistochemistry and flow cytometry, and gene expression analyses were performed by microarray techniques. RESULTS: Breast CSCs-like colonised the human-implanted bone, resulting in bone remodelling. Bone metastatic lesions were histologically apparent by tumour cell expression of epithelial markers and vimentin. The bone-isolated CSCs-like were CD44⁻CD24⁺ and showed tumorigenic abilities after injection in secondary mice. CD44⁻CD24⁺ CSCs-like displayed a distinct bone tropism signature that was enriched in genes that discriminate bone metastases of breast cancer from metastases at other organs. CONCLUSION: Breast CSCs-like promote bone metastasis and display a CSCs-like bone tropism signature. This signature has clinical prognostic relevance, because it efficiently discriminates osteotropic breast cancers from tumour metastases at other sites.

JAK2V617F activating mutation is associated with the myeloproliferative type of chronic myelomonocytic leukaemia.

BACKGROUND: Chronic myelomonocytic leukaemia (CMML) is a haematopoietic malignancy with heterogeneous clinical and morphological features. It is classified in the World Health Organization myeloproliferative-myelodysplastic overlap category. JAK2(V617F) mutation can be found in a large percentage of patients with myeloproliferative neoplasms. AIMS: To investigate the association between JAK2(V617F) mutation and clinical, haematological and bone marrow histological features in CMML and to verify whether the mutation is associated with the myeloproliferative type of the disease. METHODS: 78 consecutive patients with newly diagnosed CMML from 2004 to 2008 were included in the study. JAK2(V617F) mutation was assessed using direct sequencing of exon 14 or by allele-specific PCR from total peripheral blood or bone marrow samples. RESULTS: JAK2(V617F) mutation was identified in eight cases (10.2%). All patients with the mutation presented with splenomegaly and had a significantly higher haemoglobin level and neutrophil count than patients without the mutation. All bone marrow biopsies of JAK2(V617F)-mutated CMML showed increased erythropoiesis, a marked myeloid and megakaryocytic hyperplasia with occasionally clustered megakaryocytes, and a mild or moderate (grade 1 or 2) fibrosis; six cases showed an increased number of dilated sinusoids and reactive lymphoid nodules. CONCLUSIONS: The results indicate that JAK2(V617F) mutation is associated with clinical and morphological features of the myeloproliferative type of CMML. Therefore, JAK2 mutation analysis together with bone marrow morphology could help in a more appropriate classification of the disease.

Flow cytometry in the bone marrow staging of mature B-cell neoplasms.

BACKGROUND: Even though flow cytometric (FC) analysis of bone marrow aspirates is often performed in hematolymphoid disorders at diagnosis and during disease monitoring, its role has not been defined during the staging of B-non-Hodgkin's lymphoma (B-NHL) and B-cell lymphoproliferative diseases. The goal of this study was to provide an objective evaluation of how FC might help in the detection of bone marrow involvement by the different types of B-cell malignant neoplasms. METHODS: Fifty-four staging and 156 restaging bone marrow biopsies and bone marrow aspirates, obtained from 185 consecutive patients, were analyzed retrospectively. The results of the morphologic examination and FC were reviewed independently, and their ability to detect bone marrow involvement was compared. RESULTS: FC and morphology agreed in 176 cases (83.8%), i.e., both showed 77 positive cases and 99 negative ones. Discrepant results were obtained in 30 cases (14.2%) in which morphologic examination showed 25 (11.9%) positive cases, whereas FC showed no evidence of disease. FC detected involvement in five cases (2.4%) in the presence of a histologically negative bone marrow biopsy. All morphologically undetermined bone marrow cases (four) were negative by FC. CONCLUSIONS: Neither morphologic examination nor FC alone is adequate for the detection of all cases of B-lymphoid neoplasm bone marrow involvement. FC failed to detect bone marrow involvement in those B-NHL cases having focal paratrabecular infiltration, but proved to be more sensitive than histology in detecting small clonal B-cells in B-NHL, which demonstrated fewer than 5% neoplastic infiltrates. The clinical relevance of minimal disease detected by FC alone needs further evaluation because staging of lymphomas currently is based only on morphologic data.

A randomised, double blind, placebo controlled study of celecoxib, a selective cyclooxygenase 2 inhibitor, on duodenal polyposis in familial adenomatous polyposis.

BACKGROUND: Non-selective cyclooxygenase (COX) inhibitors (non-steroidal anti-inflammatory drugs) inhibit large bowel carcinogenesis in patients with familial adenomatous polyposis (FAP). Their role in the duodenum of these patients is less certain. The disease modifying activity of specific COX-2 inhibitors has not been explored in humans. PATIENTS AND METHODS: This was a randomised, double blind, placebo controlled study of celecoxib (100 mg twice daily (n=34) or 400 mg twice daily (n=32)) versus placebo (n=17), given orally twice daily for six months to patients with FAP. Efficacy was assessed qualitatively by blinded review of shuffled endoscopy videotapes comparing the extent of duodenal polyposis at entry and at six months and quantitatively by measurement of the percentage change in duodenal area covered by discrete and plaque-like adenomas from photographs of high and low density polyposis. RESULTS: Shuffled and blinded video review showed a statistically significant effect of 400 mg twice daily celecoxib compared with placebo treatment (p=0.033) with all five independent observers scoring a beneficial effect. Overall, patients taking celecoxib 400 mg twice daily showed a 14.5% reduction in involved areas compared with a 1.4% for placebo (p=0.436). However, patients with clinically significant disease at baseline (greater than 5% covered by polyps) showed a 31% reduction in involved areas with celecoxib 400 mg twice daily compared with 8% on placebo (p=0.049). CONCLUSIONS: A panel of five endoscopists found a significant reduction in duodenal polyposis after six months of treatment with celecoxib 400 mg twice daily. COX-2 inhibition may help this otherwise untreatable condition.

The effect of celecoxib, a cyclooxygenase-2 inhibitor, in familial adenomatous polyposis.

BACKGROUND: Patients with familial adenomatous polyposis have a nearly 100 percent risk of colorectal cancer. In this disease, the chemopreventive effects of nonsteroidal antiinflammatory drugs may be related to their inhibition of cyclooxygenase-2. METHODS: We studied the effect of celecoxib, a selective cyclooxygenase-2 inhibitor, on colorectal polyps in patients with familial adenomatous polyposis. In a double-blind, placebo-controlled study, we randomly assigned 77 patients to treatment with celecoxib (100 or 400 mg twice daily) or placebo for six months. Patients underwent endoscopy at the beginning and end of the study. We determined the number and size of polyps from photographs and videotapes; the response to treatment was expressed as the mean percent change from base line. RESULTS: At base line, the mean (+/-SD) number of polyps in focal areas where polyps were counted was 15.5+/-13.4 in the 15 patients assigned to placebo, 11.5+/-8.5 in the 32 patients assigned to 100 mg of celecoxib twice a day, and 12.3+/-8.2 in the 30 patients assigned to 400 mg of celecoxib twice a day (P=0.66 for the comparison among groups). After six months, the patients receiving 400 mg of celecoxib twice a day had a 28.0 percent reduction in the mean number of colorectal polyps (P=0.003 for the comparison with placebo) and a 30.7 percent reduction in the polyp burden (the sum of polyp diameters) (P=0.001), as compared with reductions of 4.5 and 4.9 percent, respectively, in the placebo group. The improvement in the extent of colorectal polyposis in the group receiving 400 mg twice a day was confirmed by a panel of endoscopists who reviewed the videotapes. The reductions in the group receiving 100 mg of celecoxib twice a day were 11.9 percent (P=0.33 for the comparison with placebo) and 14.6 percent (P=0.09), respectively. The incidence of adverse events was similar among the groups. CONCLUSIONS: In patients with familial adenomatous polyposis, six months of twice-daily treatment with 400 mg of celecoxib, a cyclooxygenase-2 inhibitor, leads to a significant reduction in the number of colorectal polyps.

Pure alveolar rhabdomyosarcoma of the corpus uteri: description of a case with increased serum level of CA-125.

We report a morphological and immunohistochemical study of a case of pure alveolar rhabdomyosarcoma of the uterus in an 80-year-old woman. The diagnostic clues were the characteristic "alveolar" pattern of growth, the evidence of cross-striations in strap or elongated cells with abundant eosinophilic cytoplasms, the presence of multinucleated cells with peripherally placed "wreathlike" nuclei, and the expression of muscular antigens by the tumor cells. A thorough sampling of the tumor excluded areas of other types of heterologous or homologous sarcomas or the presence of coexisting adenoma or carcinoma. The other immunohistochemical data showed a high proliferative rate as well as a high rate of p53 overexpression in the small poorly differentiated rhabdomyoblasts. Interestingly, the large differentiated rhabdomyoblasts expressed CA-125, the antigenic determinant of nonmucinous epithelial ovarian tumors. The clinical course was very aggressive: the patient died 5 months after surgery because of disease progression. The pertinent literature is discussed.

0.3% ciprofloxacin ophthalmic ointment in the treatment of bacterial keratitis. The Ciprofloxacin Ointment/Bacterial Keratitis Study Group.

OBJECTIVE: To determine the efficacy and safety of topical 0.3% ciprofloxacin hydrochloride ophthalmic ointment in the treatment of bacterial keratitis. DESIGN: Prospective case series with a nonrandomized comparison of culture-positive, evaluable cases (ciprofloxacin ointment group) with culture-positive, concurrent patients (nonenrolled group) treated with conventional therapy. SETTING: Multicenter clinical study. PATIENTS: After informed consent was obtained, 253 eligible patients underwent corneal scrapings and received topical ciprofloxacin ointment; 145 (57%) had positive cultures and completed the follow-up schedule. Forty (70%) of 57 apparently eligible patients had culture-positive bacterial keratitis but were not enrolled in the ciprofloxacin ointment study during the same period. INTERVENTION: Ciprofloxacin ophthalmic ointment instilled every 1 to 2 hours for 2 days, then every 4 hours for 12 days. MAIN OUTCOME MEASURES: Clinical evaluation of signs at 1, 3, 7, and 14 days of treatment and the overall condition classified as clinical success (cured or improved) or failure (unchanged or worse) during and after therapy. RESULTS: Clinical success with the initial treatment occurred in 135 patients (93%) in the ciprofloxacin ointment group and in 28 patients (70%) in the nonenrolled group. Of the 10 ciprofloxacin clinical failures, seven were staphylococcal; two, pneumococcal; and one, polybacterial. The 90% minimum inhibitory concentration of ciprofloxacin was 3 mg/L or less for corneal bacterial isolates. No serious adverse event attributable to ciprofloxacin ointment occurred, although 32 (13%) of 253 patients developed a transient white crystalline corneal precipitate shown with liquid chromatography in two cases to be ciprofloxacin. CONCLUSION: Ciprofloxacin ophthalmic ointment is an effective and safe topical antimicrobial agent for the treatment of bacterial keratitis caused by susceptible microorganisms.

[Adrenal neuroblastoma in adults: a rare neoplasm of difficult diagnostic interpretation. A case report]. / Il neuroblastoma del surrene nell'adulto: una rara neoplasia di difficile interpretazione diagnostica. Contributo casistico.

A case of a 25-year old girl, affected by an adrenal neoplasia with bone marrow and splenic metastases, is reported. The site of the neoplasm, that was highly undifferentiated, the presence of rosette-forming cells along with the clinical picture and the rapidity of the final outcome, could suggest the diagnosis of neuroblastoma. However we stress that the diagnosis is very difficult and can only be stated on probability bases in such undifferentiated neoplasms.

Trisomy 8 and an unbalanced t(5;17)(q11;p11) characterize two karyotypically independent clones in a case of idiopathic myelofibrosis evolving to acute nonlymphoid leukemia.

In a patient with idiopathic myelofibrosis (MFI) that had progressed to acute nonlymphoid leukemia (ANLL) after a long-lasting cytotoxic treatment, we observed two karyotypically independent cell populations, one showing trisomy of chromosome 8 as the only anomaly and one with an unbalanced translocation t(5;17)(q11) resulting in partial monosomy of 5q and 17p. The overall karyotypic configuration suggested that chromosome changes occurred as secondary events during the multistep process of leukemogenesis. The probable sequence of cytogenetic events in this patient and a review of the literature indicated that the t(5;17) may represent a therapy-induced abnormality nonrandomly related to the terminal phase of myeloid disorders.

Immunophenotype of thymoma-associated lymphoid cell component of T-cell type. A new analytic procedure in keeping with structural heterogeneities.

The phenotype of the lymphoid cell component of 35 thymomas was investigated by analyzing cryostat sections and lymphocyte suspensions. The morphology in each case was determined by examining multiple tissue samples from different parts of the tumor. Structural heterogeneity was shown in 14 thymomas, and a homogeneous morphology of cortical or medullary or mixed types in the others. To assess whether this heterogeneity was correlated with differences in the lymphoid phenotype, we analyzed both lymphocyte suspensions and frozen sections from the same samples. Phenotypical differences in the suspensions of each thymoma in the heterogeneous group were noted and similar differences were also observed in the cryostat sections. Phenotypical abnormalities were found in some thymomas. They consisted of the simultaneous expression of cortical and medullary markers, which was most marked in the heterogenous mixed-type thymomas invading the lung. Furthermore, the global phenotype was tested on a pool of lymphocyte suspensions in all thymomas. This procedure distinguished cortical, medullary and intermediate cortico-medullary immunophenotype models which closely correlated with the tumor histology. It was concluded that, due to the frequent structural and immunological heterogeneity of thymomas, correct assessment of their lymphoid component requires a two-step analysis. This comprises: 1) individual suspensions from samples taken from different areas of the same thymoma, and 2) a pool of these suspensions. The first step will reveal the different immunological characteristics. In the second, the lymphocyte phenotype, which may vary widely throughout the tumor, will be represented in its totality. These findings may be of great help in predicting clinical patterns, especially possible malignant evolution.

Aspheric rigid gas permeable contact lenses: practitioner discrimination of base curve increments using fluorescein pattern evaluation.

Evaluation of the fluorescein pattern is the main clinical indicator of the fit of a rigid gas permeable (RGP) contact lens to the cornea. Traditionally, manufacturers inventory RGP lenses in 0.05-mm base curve increments. The objective of this study was to test the hypothesis that experienced practitioners are not able to discern base curve/cornea relations of aspheric design lenses correctly to within +/- 0.05 mm. Eleven clinicians with expertise in fitting rigid lenses were asked to interpret fluorescein patterns and lens fittings of the Bausch & Lomb Quantum aspheric RGP lens on five adapted rigid lens wearers. The results showed that the experienced practitioners used in this study were not able to discriminate cornea/base curve relations of Quantum aspheric lenses within +/- 0.05 mm more than 67% of the time (p less than 0.05). There were no patient-to-patient differences based on type of lens fit (interpalpebral or lid attachment) or degree of corneal toricity. Lenses fitted in 0.10-mm base curve steps were judged as being as acceptable as lenses fitted in 0.05 mm steps (p less than 0.05). It was concluded that Quantum aspheric design lenses need only be trial fitted in 0.10-mm base curve steps.

Multiple applications of the NCT: an assessment of the instrument's effect on IOP.

The feasibility of the American Optical Non-Contact Tonometer as an instrument for tonography was assessed. It was found that after 20 measurements in quick succession, there was a statistically insignificant change in IOP. Multiple intraocular pressure (IOP) readings with this instrument are helpful in determining an abnormally high IOP from an aberrant recording.

Factors influencing vision with rigid gas permeable alternating bifocals.

This study assessed the fitting and visual performance of a rigid, gas permeable (RGP), monocentric, alternating bifocal. Fourteen presbyopic eyes each wore 24 different lenses consisting of all combinations of 2 diameters, 2 segment heights, 2 prism ballasts, and 3 fitting relations. The influence of parameter selection on visual performance and the usefulness of clinical measurements in predicting visual performance were determined statistically by multivariate logistic regression. Our results showed that prism, segment height, and the fitting relation can influence distance and near visual performance. Lens movement, postblink segment positioning, and return time were the most useful measured predictors of visual performance.

Immunologic cell markers of peripheral blood and lymph node lymphocytes in chronic lymphocytic leukemia.

The immunologic phenotype of the lymphocytes of 100 patients with chronic lymphocytic leukaemia (CLL) was investigated. Peripheral blood lymphocytes (PBL) were examined in all cases; in 46 patients with lymphadenopathy, a lymph node was biopsied and the histologic and immunologic patterns were assessed: 24 had a lymphocytic-lymphoplasmocytoid histology and 22 the follicular variant of lymphocytic lymphoma (mantle zone lymphoma, MZL). For comparison, lymph node suspensions from 19 patients with non-leukemic centrocytic lymphoma (CCL) were also studied. Significant differences in the PBL immunologic features were found between stage O and stage I patients. The phenotype of the lymphocytes of patients with lymphocytic histology was similar to that of stage 0 CLL patients, whereas major differences were found between these patients and those with mantle zone histology. This enables these patients to be recognized easily on immunologic grounds.

Acid hydrolases in B-chronic lymphocytic leukemia (B-CLL): a comparison with normal peripheral B lymphocytes and normal B-cell subset with the phenotype of B-leukaemic cells.

The acid phosphatase (AP), beta-glucuronidase (BG), and alpha-naphthyl-acetate esterase (ANAE) distribution and the morphology of stage 0 B-CLL lymphocytes were studied. The results were compared with the same hydrolase equipment and morphology of normal B-cell populations showing the B-CLL-like phenotype, and thus regardable as the possible counterpart of leukaemic cells. The functional and structural characters of the former were strikingly different from those of the latter. In fact the normal B-cell population with B-CLL-like phenotype showed an homogeneous enzyme pattern with predominant ANAE and a strictly lymphocytic cell morphology. In contrast, the leukaemic cells showed various and unrelated morphological and cytochemical features, thus forming apparent subgroups of B-CLL. The development of an irregular "switch on" mechanism in the blocked malignant cell clone, may account for these functional and structural maturation discrepancies. Moreover, the fact that in the leukaemic cells ANAE activity could be demonstrated only after the appearance of the other hydrolases, makes it a marker of functionally differentiated B lymphocytes.

Deswelling of the cornea after hypoxia.

Central corneal thickness changes were monitored in nine subjects who wore low water content hydrogel lenses of varying thickness for 3 h in the open and closed eye conditions. At the end of 3 h of lens wear, corneal deswelling to prelens thickness for the different levels of hypoxia was determined by pachometry performed every 5 min for the first 30 min, every 10 min for the next 40 min, and every 15 min thereafter. Examination of the individual deswelling profiles indicates (1) a biphasic response in those with corneal edema greater than 14%, and (2) a large portion of the deswelling occurred during the first 30 min, followed by a more gradual rate until baseline was reached. Small amounts of corneal edema (less than 5%) required up to 25 min to return to baseline, whereas 6 to 15% required 30 to 70 min, and 20% required 95 min. The time taken for the cornea to deswell to baseline was linearly related to the amount of corneal swelling (r = 0.88; p less than 0.0001).

Clinical prediction of postsurgical aphakic refractive correction.

A simple clinical predictor of the amount of spectacle convex lens power that will be needed to correct the refractive error of an aphakic patient after cataract surgery is developed based on clinical measures of anterior corneal surface power (keratometer measurements) and axial length. This predictor derived from matrix optics is compared to previous ones developed by Binkhorst and Sanders et al. using different methodologies.

Clinical determination of the corrected retinal image size in spectacle-corrected aphakes.

For individual spectacle-corrected aphakic patients, a method of estimating the size of the corrected retinal image corresponding to distant objects smaller than Snellen 6/360 (20/1200) is developed. Our results indicate that an estimate of the corrected retinal image size corresponding to a distant Snellen 6/12 (20/40) object in an individual spectacle-corrected aphake can be obtained to within plus or minus 1.0 micron of error using a multivariate regression model based on the measurement of six patient factors: axial length, anterior corneal power, spectacle lens back vertex power, lens thickness, lens back curve, and lens vertex distance. Clinical application of these results are discussed along with comparisons to other results based on traditional spectacle magnification formulas. The size of the corrected retinal image corresponding to a Snellen 6/12 (20/40) object was found to vary between 60 and 84 micron in patients corrected with CR-39 optical plastic and for patients corrected with high-index optical glass, it was between 56 and 78 microns.