Soybean-associated endophytic fungi as potential source for anti-COVID-19 metabolites supported by docking analysis.

AIMS: To identify the metabolites produced by the endophytic fungus, Aspergillus terreus and to explore the anti-viral activity of the identified metabolites against the pandemic disease COVID-19 in-silico. METHODS AND RESULTS: Herein, we reported the isolation of A. terreus, the endophytic fungus associated with soybean roots, which is then subcultured using OSMAC approach in five different culture media. Analytical analysis of media ethylacetate extracts using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) was carried out. Furthermore, the obtained LC-MS data were statistically processed with MetaboAnalyst 4.0. Molecular docking studies were performed for the dereplicated metabolites against COVID-19 main protease (Mpro ). Metabolomic profiling revealed the presence of 18 compounds belonging to different chemical classes. Quinones, polyketides and isocoumarins were the most abundant classes. Multivariate analysis revealed that potato dextrose broth and modified potato dextrose broth are the optimal media for metabolites production. Molecular docking studies declared that the metabolites, Aspergillide B1 and 3a-Hydroxy-3, 5-dihydromonacolin L showed the highest binding energy scores towards COVID-19 main protease (Mpro ) (-9·473) and (-9·386), respectively, and they interact strongly with the catalytic dyad (His41 and Cys145) amino acid residues of Mpro . CONCLUSIONS: A combination of metabolomics and in-silico approaches have allowed a shorter route to search for anti-COVID-19 natural products in a shorter time. The dereplicated metabolites, aspergillide B1 and 3α-Hydroxy-3, 5-dihydromonacolin L were found to be potent anti-COVID-19 drug candidates in the molecular docking study. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed that the endophytic fungus, A. terreus can be considered as a potential source of natural bioactive products. In addition to, the possibility of developing the metabolites, aspergillide B1 and 3α-Hydroxy-3, 5-dihydromonacolin L to be used as phytopharmaceuticals for the management of COVID-19.

Non-infectious Pulmonary Diseases and HIV.

Pulmonary complications remain among the most frequent causes of morbidity and mortality for individuals with HIV despite the advent of antiretroviral therapy (ART) and improvement in its efficacy and availability. The prevalence of non-infectious pulmonary diseases is rising in this population, reflecting both an increase in smoking and the independent risk associated with HIV. The unique mechanisms of pulmonary disease in these patients remain poorly understood, and direct effects of HIV, genetic predisposition, inflammatory pathways, and co-infections have all been implicated. Lung cancer, chronic obstructive pulmonary disease (COPD), and pulmonary hypertension are the most prevalent non-infectious pulmonary diseases in persons with HIV, and the risk of each of these diseases is higher among HIV-infected (HIV+) persons than in the general population. This review discusses the latest advances in the literature on these important complications of HIV infection.

Serum YKL-40 in psoriasis with and without arthritis; correlation with disease activity and high-resolution power Doppler ultrasonographic joint findings.

BACKGROUND: YKL-40 may be involved in angiogenesis in psoriasis and psoriatic arthritis (PsA). High-resolution power Doppler ultrasound (PDUS) can detect angiogenesis of synovium in PsA. AIM: To assess serum YKL-40 in psoriasis patients with or without PsA, and to correlate its levels with disease activity and high-resolution PDUS findings. METHODS: In this case-control study, 48 patients with psoriasis (26 of them also had PsA) and 30 controls were assessed by high-resolution PDUS, and assayed for serum levels of YKL-40 by ELISA. Patients were clinically assessed using Composite Psoriatic Disease Activity Index (CPDAI). Total joint score (TJS) was used to assess joint involvement in PsA. RESULTS: A statistically significant elevation was found in YKL-40 levels in psoriatics with or without PsA compared with controls (P < 0.001), as well as in PsA (group II) compared to patients without arthritis (group I) (P = 0.002). CPDAI, synovial thickness score and colour Doppler ultrasound (CDUS) score were highly significantly higher in group II vs. group I (P < 0.001). In all patients, CPDAI, synovial thickness and CDUS score were positively correlated to each other, and each of them was positively correlated to serum YKL-40 levels (P < 0.05). In either group I or II, serum YKL-40 levels correlated positively with CPDAI (P < 0.05). In group II, TJS, synovial thickness and CDUS score were positively correlated to each other (P < 0.05). CONCLUSIONS: Serum YKL-40 can be used as a new biological marker for angiogenesis and disease activity in psoriasis with or without PsA. High-resolution PDUS is a non-invasive tool for the evaluation of angiogenesis in PsA patients as well as for the detection of early synovial changes in psoriasis patients without arthritis.

Platelet activation: a link between psoriasis per se and subclinical atherosclerosis--a case-control study.

BACKGROUND: Pathomechanisms of both psoriasis and atherosclerosis may involve platelet activation. Activated platelets show increased P-selectin; CD62 expression, and mean platelet volume (MPV). Impaired brachial artery flow-mediated dilatation (FMD) is related to atherosclerosis. OBJECTIVES: To determine the presence of subclinical atherosclerosis in patients with psoriasis (without overt cardiovascular complications or traditional cardiovascular disease risk factors), compared with controls. METHODS: In this case-control study, 25 patients with psoriasis and 25 age- and gender-matched healthy individuals were subjected to assessment of MPV, CD62 expression using flow cytometry, and brachial artery FMD and transthoracic echocardiography by cardiac ultrasound scanner. RESULTS: A statistically highly significant increased CD62 expression, but not MPV, was found in cases compared with controls, and in patients with moderate/severe psoriasis compared with either mild cases or controls (P < 0.001). CD62 expression was statistically significantly positively correlated with the Psoriasis Area and Severity Index (PASI) score (P < 0.001), baseline brachial artery diameter (P = 0.03) but not FMD and aortic root diameter (ARD; P = 0.03). ARD was statistically significantly higher in patients with moderate/severe psoriasis compared with controls (P = 0.017). Stepwise simple linear regression analysis revealed that PASI score was the most important factor affecting CD62 expression (P < 0.001). CONCLUSIONS: Our study showed increased atherosclerosis risk in patients with psoriasis, particularly those with moderate/severe disease, as evidenced by increased expression of platelet CD62 compared with healthy controls. Moreover, we found a positive correlation between CD62 expression and ARD (another possible marker of atherosclerosis), with positive correlation to the PASI score; the most important factor influencing CD62 expression. However, our data on MPV and FMD do not support the use of either value for diagnosing subclinical atherosclerosis in patients with psoriasis in further studies.

Olanzapine versus placebo for out-patients with anorexia nervosa.

BACKGROUND: Anorexia nervosa (AN) is a serious psychiatric illness associated with significant morbidity and mortality. There is little empirical support for specific treatments and new approaches are sorely needed. This two-site study aimed to determine whether olanzapine is superior to placebo in increasing body mass index (BMI) and improving psychological symptoms in out-patients with AN. METHOD: A total of 23 individuals with AN were randomly assigned in double-blind fashion to receive olanzapine or placebo for 8 weeks together with medication management sessions that emphasized compliance. Weight, other physical assessments and measures of psychopathology were collected. RESULTS: End-of-treatment BMI, with initial BMI as a covariate, was significantly greater in the group receiving olanzapine [F(1, 20)=6.64, p=0.018]. Psychological symptoms improved in both groups, but there were no statistically significant group differences. Of the 23 participants, 17 (74%) completed the 8-week trial. Participants tolerated the medication well with sedation being the only frequent side effect and no adverse metabolic effects were noted. CONCLUSIONS: This small study suggests that olanzapine is generally well tolerated by, and may provide more benefit than placebo for out-patients with AN. Further study is indicated to determine whether olanzapine may affect psychological symptoms in addition to BMI.

Anterior cruciate ligament tear patterns in young patients: An arthroscopic multicenter study.

INTRODUCTION: In the last decade, there has been a renewed interest in anterior cruciate ligament (ACL) preservation surgeries in the younger patients. Several ACL preservation techniques such as primary repair, augmented repair, and scaffold repair have been described based on the particular tear type and pattern. The purpose of this study was to determine the distribution of tear patterns in young patients presenting with an acute ACL injury. METHODS: A prospective observational study was performed at two tertiary children's hospitals. Patients under 18 years undergoing ACL reconstruction within 8 weeks of initial injury were included from 2017 to 2019. Tear patterns were classified by two orthopedic surgeons from each of the two centers during arthroscopic ACL reconstruction into 4 types: I. Avulsion off the femur, II. <10% of total ACL length tear from femoral end, III. Mid-substance tear and IV. Single bundle tear. For reliability, the four surgeons classified ACL injury (2 rounds each) based on de-identified intraoperative videos of 33 randomly selected surgical ACL cases. Inter and intra-rater reliability studies were calculated using Kappa statistics. RESULTS: 224 patients (123 males, 101 females) with mean age of 16 (range: 9-18) years were enrolled in this study. Fifty-seven (25%) patients reported contact injury while 167 (75%) reported non-contact. Isolated ACL injury was recorded in 70 (31%) patients, while concomitant injuries were recorded in 154 patients (69%). The most common associated injury was lateral meniscus tear (35%), followed by lateral and medial meniscus tears (20%). According to our classification, 31 (14%) patients were Type I, 30 (13%) were Type II, 139 (62%) were Type III, 18 (8%) were Type IV. The intra-rater reliability was excellent for 2 reviewers, good for 1 and marginal for another. The overall inter-rater reliability for all 4 reviewers was marginal for both readings. There was no statistical difference in the occurrence of type of tear based on the mechanism of injury (contact vs non-contact) or age of the patients. CONCLUSIONS: This is the first multicenter study using an arthroscopic assessment to classify the location of ACL tear in the young population. It gives us further insight on the possible application for surgeries to preserve the ACL in this group. Larger studies incorporating these findings with MRI evaluation and ACL repair techniques are needed to confirm the utility of this information to decide the eligibility for repair in pediatric patients.

Upper gastrointestinal findings and detection of Helicobacter pylori in patients with oral lichen planus.

BACKGROUND: Lichen planus (LP) is a mucocutaneous disease of unknown aetiology, which may involve the gastrointestinal (GI) mucosa. The association of Helicobacter pylori with LP has been a subject of debate. AIM: To investigate upper GI findings and the presence of H. pylori in GI mucosa and oral LP (OLP). METHODS: Oral biopsies from 20 patients with erosive OLP and 20 with non-erosive OLP were investigated for the presence of H. pylori by histopathological examination and PCR. Upper GI endoscopy and GI mucosal biopsies were examined for LP lesions and/or H. pylori. RESULTS: The endoscopic findings of both groups were oesophagitis, antral gastritis and duodenitis. No LP or LP-like changes were found in the upper GI mucosa. H. pylori was found by histopathological examination in the gastric mucosa of 18 patients (45%), with equal distribution in both the control and study groups. Positive PCR results were obtained from biopsy specimens of oral lesions in all patients with erosive OLP and presence of H. pylori in the stomach (9 patients), but in none of the patients with non-erosive OLP (P = 0.001). CONCLUSION: We did not find any difference in symptoms, endoscopic findings and histopathological results between patients with erosive and non-erosive OLP. However, the concomitant presence of erosive OLP, of H. pylori nucleic acid in erosive OLP and the H. pylori organisms in gastric mucosa implies a possible pathogenic connection between this bacterium and erosive OLP.

The slippery slope: prediction of successful weight maintenance in anorexia nervosa.

BACKGROUND: Previous research has found that many patients with anorexia nervosa (AN) are unable to maintain normal weight after weight restoration. The objective of this study was to identify variables that predicted successful weight maintenance among weight-restored AN patients. METHOD: Ninety-three patients with AN treated at two sites (Toronto and New York) through in-patient or partial hospitalization achieved a minimally normal weight and were then randomly assigned to receive fluoxetine or placebo along with cognitive behavioral therapy (CBT) for 1 year. Clinical, demographic and psychometric variables were assessed after weight restoration prior to randomization and putative predictors of successful weight maintenance at 6 and 12 months were examined. RESULTS: The most powerful predictors of weight maintenance at 6 and 12 months following weight restoration were pre-randomization body mass index (BMI) and the rate of weight loss in the first 28 days following randomization. Higher BMI and lower rate of weight loss were associated with greater likelihood of maintaining a normal BMI at 6 and 12 months. An additional predictor of weight maintenance was site; patients in Toronto fared better than those in New York. CONCLUSIONS: This study found that the best predictors of weight maintenance in weight-restored AN patients over 6 and 12 months were the level of weight restoration at the conclusion of acute treatment and the avoidance of weight loss immediately following intensive treatment. These results suggest that outcome might be improved by achieving a higher BMI during structured treatment programs and on preventing weight loss immediately following discharge from such programs.

A minimally interactive and reproducible method for abdominal aortic aneurysm quantification in 3D ultrasound and computed tomography with implicit template deformations.

The maximum diameter of abdominal aortic aneurysm (AAA) is a key quantification parameter for disease assessment. Although it is routinely measured on 2D-ultrasound images, using a volumetric approach is expected to improve measurement reproducibility. In this work, 3D-ultrasound or computed tomography imaging of patients with AAA was combined with a minimally interactive 3D segmentation based on implicit template deformation. Segmentation usability and reproducibility were evaluated on 81 patients, showing a mean measurement time of [2;8]min per case, and Dice coefficients of 0.87±0.12 for 3D-US and 0.81±0.08 for CT. Quantification parameters included a diameter measurement from 3D-US and CT volumes with respective confidence intervals of 0.51 [-2.5;3.52]mm and 1.00 [-1.68;3.67]mm. Additional volume measurements showed confidence intervals of 0.91 [-4.17;5.99]ml for 3D-US and 4.10 [-4.11;12.30]ml for CT.

An open trial of an intensive summer day treatment program for severely overweight adolescents.

This open trial examined the feasibility and usefulness of treating adolescents with a body mass index (BMI) > or =95th percentile in a 6-week day treatment program within a psychiatric outpatient setting. Sixteen adolescents, ages 12-15, attended a 6-week multidisciplinary summer day treatment program. Outcome measures include pre- and post-program assessments of BMI, body fat, laboratory measures, nutritional status, physical activity, mood and eating disorder symptoms, motivation, self-esteem and quality of life. BMI, waist/hip circumferences, blood pressure, heart rate, and body fat did not change significantly from baseline to week 6. Fasting glucose decreased significantly from baseline to week 6, but there were no significant changes in other laboratory measures. The teen-rated feelings subscale of the Peds Quality of Life Scale improved. Self-esteem, as measured by the Rosenberg Self- Esteem Scale, indicated a strong trend towards improvement, and motivation measures also showed a trend for improvement. These findings suggest that a brief, intensive intervention for overweight adolescents may yield meaningful changes in quality of life, self-esteem, and motivation, even in the absence of significant weight loss.

Intracellular calcium response of ACL and MCL ligament fibroblasts to fluid-induced shear stress.

This study examines the real-time intracellular calcium concentration, [Ca2+]i, response of canine medial collateral ligament (MCL) and anterior cruciate ligament (ACL) fibroblasts subjected to a fluid-induced shear stress of 25 dynes/cm2. In experiments using a modified Hanks' Balanced Salt Solution (HBSS) perfusate, both cell types demonstrated a significant increase in peak [Ca2+]i compared to respective no-flow controls, the response of MCL fibroblasts being nearly 2-fold greater than that of ACL fibroblasts. In studies where the cells were bathed in a medium of HBSS supplemented with 2% newborn bovine serum (NBS) and then introduced to flow with the same medium, ACL fibroblasts responded nearly 3-fold greater than MCL fibroblasts. Neomycin (10 mM), thapsigarigin (1 microM) and Ca(2+)-free media supplemented with EGTA (1 mM) were able to inhibit significantly the [Ca2+]i response to flow with HBSS in both fibroblasts. Thapsigargin also blocked the NBS flow response in both cell types, while neomycin and Ca(2+)-free media significantly inhibited the ACL response. Our findings demonstrate that ACL and MCL cells are not the same. These differences may be related to the disparate healing capacity of the ACL and MCL observed clinically.

Comparative efficacy of selective serotonin reuptake inhibitors and tricyclics in the treatment of melancholia.

OBJECTIVE: The popularity of selective serotonin reuptake inhibitors stems from their apparent efficacy for numerous disorders and their favorable side effect profile. However, several studies have suggested that selective serotonin reuptake inhibitors may be relatively ineffective for treating melancholia. The objective of this study was to compare the responses to fluoxetine and nortriptyline of older patients with both severe depression and heart disease. METHOD: The outcome of 22 hospitalized patients with unipolar depression and heart disease who were treated with fluoxetine was compared to the outcome of 42 comparable patients treated with nortriptyline. The average age of the fluoxetine group was 73 years, and their mean pretreatment score on the Hamilton Depression Rating Scale was 26; the average age of the nortriptyline group was 70, and their mean pretreatment Hamilton score was 28. RESULTS: Of the 42 nortriptyline-treated patients, 28 were responders, six were nonresponders, and eight dropped out. The intent-to-treat response rate was 67% (28 of 42), and the response rate of the melancholic patients who completed the nortriptyline trial was 83% (20 of 24). Of the 22 fluoxetine-treated patients, five were responders, 13 were nonresponders, and four dropped out. The intent-to-treat response rate was 23% (five of 22), and the response rate of the melancholic patients who completed the fluoxetine trial was 10% (one of 10). CONCLUSIONS: Fluoxetine appears to be significantly less effective than nortriptyline for treating hospitalized elderly patients with unipolar major affective disorder, especially those with the melancholic subtype and concurrent cardiovascular disease.

Does fluoxetine augment the inpatient treatment of anorexia nervosa?

OBJECTIVE: While pharmacological interventions are of established utility in bulimia nervosa, medications have no clear role in the treatment of anorexia nervosa. Because patients with anorexia nervosa frequently exhibit mood disturbances and symptoms of obsessive-compulsive disorder, the authors tested the utility of fluoxetine in the treatment of women participating in an inpatient program for anorexia nervosa. METHOD: The authors conducted a randomized, placebo-controlled, double-blind, 7-week study of fluoxetine at a target daily dose of 60 mg in 31 women with anorexia nervosa receiving treatment for their eating disorder on a clinical research unit. Body weight and measures of eating behavior and psychological state were obtained at baseline and at termination. RESULTS: There were no significant differences in clinical outcome on any measure between patients receiving fluoxetine and patients receiving placebo. CONCLUSIONS: Fluoxetine does not appear to add significant benefit to the inpatient treatment of anorexia nervosa.

Cardiovascular effects of fluoxetine in depressed patients with heart disease.

OBJECTIVE: The purpose of this study was to determine the cardiovascular effects of fluoxetine in depressed patients with cardiac disease. METHOD: Twenty-seven depressed patients (26% of whom were female and whose average age was 73 years) who had congestive heart failure, conduction disease, and/or ventricular arrhythmia were studied in an open medication trial of fluoxetine, up to 60 mg/day, for 7 weeks. The main outcome measures were heart rate and rhythm measured by 24-hour ECG recordings, ejection fraction determined by radionuclide angiography, cardiac conduction intervals, and blood pressure. Baseline values were compared with those at weeks 2 and 7 of fluoxetine treatment. In 60 comparable patients, values of these same cardiovascular measures at baseline and after 3 weeks of treatment with a tricyclic antidepressant, nortriptyline, were also examined. RESULTS: Fluoxetine induced a statistically significant 6% decrease in heart rate, a 2% increase in supine systolic pressure, and a 7% increase in ejection fraction. There was no effect on cardiac conduction, ventricular arrhythmia, or orthostatic blood pressure. Overall, 4% of the fluoxetine patients had an adverse cardiovascular effect. In contrast, nortriptyline treatment caused a significant increase in heart rate and orthostatic hypotension, and 20% of the nortriptyline-treated patients had an adverse cardiovascular effect. CONCLUSIONS: In depressed patients with heart disease, fluoxetine treatment was not associated with the cardiovascular effects documented for the tricyclic antidepressants or with significant adverse cardiac events. However, limited conclusions about fluoxetine's cardiovascular effects and safety can be drawn from this study of only 27 patients monitored for 7 weeks.

Low levels of transthyretin in the CSF of depressed patients.

OBJECTIVE: Transthyretin plays an important role in the transport and distribution of thyroid hormone in the central nervous system (CNS). This study replicated and extended to patients with nonrefractory depressive illness a pilot study indicating that patients with refractory major depression have significantly lower levels of CSF transthyretin than do healthy comparison subjects. METHOD: Lumbar punctures were performed in drug-free subjects with DSM-III-R major depression (N = 18), DSM-III-R bipolar disorder, depressed phase (N = 1), and healthy comparison subjects (N = 24). CSF concentrations of transthyretin, determined by a quantitative dot-immunobinding assay, of the depressed patients and comparison subjects were compared by analysis of covariance (ANCOVA). The relationship between CSF transthyretin levels and Hamilton Depression Rating Scale scores was determined in a subset of the depressed patients. RESULTS: CSF concentrations of transthyretin were significantly lower in the depressed patients than in the comparison subjects by ANCOVA. Within the depressed group there was no significant overall correlation between CSF transthyretin levels and Hamilton depression scale scores, but there was a significant inverse correlation in male depressed patients (N = 8) between CSF transthyretin concentrations and Hamilton depression scores. CONCLUSIONS: Lower CSF transthyretin concentrations in depressed patients may reflect either a stable trait in this population or a state change secondary to depression or other factors. Lower CSF transthyretin concentrations may result in altered CNS thyroid hormone homeostasis. Such alteration could account for certain mood and neurovegetative symptoms of depression and might contribute to failure of standard antidepressant treatment.

Effect of serum and platelet-derived growth factor on chondrocytes grown in collagen gels.

In this in vitro study, cell proliferation, viability, and morphology; proteoglycan (PG) synthesis; and gel contraction were assessed over a 15-day period (on days 3, 6, 9, 12, and 15) for mature bovine chondrocytes cultured in collagen gels. The environment within the gel was varied by changing the concentration of fetal bovine serum (1% and 10%) and platelet-derived growth factor-BB (PDGF; 0, 10, 50, 100 ng/ml) within the gel and incubation media. Our results showed that the amount of serum or PDGF added to the gels had no effect on cell viability, with >95% of cells remaining alive throughout the experiment. There was a significant increase in cell number over time in all groups, with a higher rate of cell proliferation in gels containing 10% serum and higher concentrations of PDGF. In addition, the amount of serum significantly affected gel contraction with or without PDGF. Gels containing 10% serum contracted on day 10-12, while none of the gels containing 1% serum contracted over the course of the experiment. The PG content within each gel increased with incubation time only for the gels containing 1% serum, and 10 or 100 ng/ml of PDGF. However, on a per cell basis, there was no change in the PG content with time when only serum was used and a significant decrease in the rate of PG production with the addition of PDGF (9.1-27.8 pgPG/cell/day). Cell morphology was also affected by PDGF, with the cells becoming more spindle shaped. Cell alignment within the gels appeared to be most affected by gel contraction. Collagen gels can act as cell carriers for the purpose of tissue engineering. These gels provide a three-dimensional environment in which chondrocytes can proliferate and produce matrix. We have shown how this environment can be controlled to affect gel contraction, rates of cell growth and PG production, and cellular morphology while maintaining cell viability. This information will be useful in determining the conditions in which chondrocytes can be grown within collagen gels and combined with cytokines to create an ideal tissue construct.

A new external approach to the pterygomaxillary fossa and parapharyngeal space.

A new direct approach to the area of the pterygomaxillary fossa and the parapharyngeal space is described. This procedure was developed because previously described methods either offered limited access to the area or resulted in significant functional defects. The approach described here results in a wide-field exposure of both the pterygomaxillary and parapharyngeal spaces with no sacrifice of either mandibular function or the sensory supply of the face or oral cavity. The parapharyngeal space is entered through a transcervical incision. This, combined with double osteotomies of the mandible, allows the ascending ramus with its intact neurovascular bundle to be reflected laterally and superiorly, along with the attached masseter muscle and the overlying skin. The result is an excellent exposure of the pterygomaxillary fossa and the base of skull. Following removal of the tumor, the ramus of the mandible is replaced and fixed with interosseous wiring and the application of arch bars, thus restoring normal dental occlusion. The technique described here was worked out on cadaveric dissection before being applied to a clinical case.

Characterization of chemotactic migration and growth kinetics of canine knee ligament fibroblasts.

Migration and proliferation of ligament fibroblasts are essential for the healing of ligament injuries. This study was designed to evaluate the migration of intraarticular (anterior and posterior cruciate) and extraarticular (medial and lateral collateral) ligament fibroblasts in response to cytokines and to determine the effect of cell passage on cell proliferation. Recombinant human platelet-derived growth factor, hepatocyte growth factor/scatter factor, and bone morphogenic protein-2 stimulated the migration of all ligament cells in a dose-dependent manner, with optimal migration at 10 ng/ml. Recombinant human epithelial growth factor preferentially stimulated the migration of intraarticular ligament fibroblasts, whereas recombinant human interleukin-1 was more effective with extraarticular ligament fibroblasts. Recombinant human insulin-like growth factor-1, insulin-like growth factor-2, transforming growth factor-beta, and fibroblast growth factor had no significant effect on the migration of ligament-derived fibroblasts. These data suggest that specific cytokines stimulate the migration of knee ligament fibroblasts and provide a rationale for possible therapeutic approaches to optimize ligament healing. Fibroblasts derived from the anterior cruciate ligament have been shown to proliferate at a slower rate than those derived from the medial collateral ligament. We have extended these observations and have demonstrated that fibroblasts from both the posterior and anterior cruciate ligaments proliferate at a slower rate than lateral and medial collateral ligament-derived fibroblasts. The differences between the growth rates of intraarticular and extraarticular fibroblasts become insignificant with serial passaging of the cells.

Differential expression of integrin subunits in canine knee ligament fibroblasts.

A method for measuring the expression of integrin subunits on the cell surface of knee ligament fibroblasts was developed with use of flow cytometry and immunofluorescence. The ligament cells exhibited uniform size and density, as shown by forward and side-scatter properties, and showed minimal nonspecific binding of isotype control antibodies compared with unstained cells. All cells expressed the alpha5 integrin subunit; lateral collateral ligament cells stained with antibody to alpha5 showed a mean fluorescence intensity 2-fold higher than that of medial collateral ligament cells, 1.5-fold higher than that of posterior cruciate ligament cells, and 3-fold higher than that of anterior cruciate ligament cells, indicating a greater expression of the alpha5 subunit by lateral collateral ligament cells than by medial collateral, posterior cruciate, and anterior cruciate ligament cells. All cells expressed the beta1 integrin subunit; the expression by posterior cruciate ligament cells was 3-fold higher than that by medial collateral ligament or lateral collateral ligament cells and 5-fold higher than that by anterior cruciate ligament cells. All cells expressed the beta3 integrin subunit; the expression by posterior cruciate ligament cells was 1.5, 3, and 4.5-fold greater than that by lateral collateral, anterior cruciate, and medial collateral ligament cells, respectively. Our data suggest there is a differential expression of integrin subunits in knee ligament fibroblasts, and this in part may explain differences in their attachment and adherence to extracellular matrix molecules.

Laryngeal lymphoscintigraphy.

A method of studying lymphatic drainage of the larynx was undertaken using radioactive colloids. Sites of injection were the true and false cords, aryepiglottic folds, anterior and posterior commissure, epiglottis and arytenoid. The patient was then scanned with the gamma camera 3 to 5 hours and again 24 hours post injection. Thirty-six patients were injected and results were recorded as to previous X-ray therapy, nodal activity post scanning, ipsilateral or contralateral and distant spread, and the type of radioactive particle--99mTc labeled sulfur colloid, 99mTc microalbumin (200-800 nm diameter), and 99mTc minimicroalbumin (less than 50 nm diameter). The three radiopharmaceuticals gave similar results. Previous X-ray therapy did not alter lymphatic drainage. Of 36 patients, 23 showed nodal activity on scintiscanning: none showed any axillary nor mediastinal activity.