Heterogeneity of squamous cell carcinomas of the oral cavity studied by flow cytometry

The DNA pattern was determined by flow cytometry in 76 samples from 16 squamous cell carcinomas of the oral cavity to assess intratumour DNA heterogeneity. Heterogeneous DNA content was found in 2 tumours (12%); both containing DNA diploid and DNA aneuploid cell clones. The remaining 14 tumours showed a homogeneous DNA distribution in the different specimens; 9 (56%) were diploid, 3 (19%) aneuploid and 2 (12%) were polyploid. The DNA non-diploid tumours were clinically more advanced than the DNA diploid ones (p less than 0.05). The tumour proliferation rate (fraction of cells in S-phase) was higher in aneuploid tumours than in diploid ones (p less than 0.01).     

Cardiovascular morbidity and the use of inhaled bronchodilators

We used the Manitoba Health database to examine the relationship between use of inhaled respiratory drugs in people with chronic obstructive respiratory diseases and cardiovascular hospitalizations from 1996 through 2000. The drugs examined were beta agonists [BA], ipratropium bromide IB, and inhaled steroids (ICS). End points were first hospitalizations for supraventricular tachycardia, myocardial infarction, heart failure or stroke. A nested case control analysis was employed comparing people with and without cardiovascular events. Cases and controls were matched for gender and age, and conditional logistic regression was used in multivariate analysis considering other respiratory drugs, respiratory diagnosis and visit frequency, non-respiratory, non-cardiac comorbidities, and receipt of drugs for cardiovascular disease.In univariate analyses, BA, IB and ICS were all associated with hospitalizations for cardiovascular disease, but in multivariate analyses ICS did not increase risk while both BA and IB did. There were interactions between respiratory and cardiac drugs receipt in that bronchodilator associated risks were higher in people not taking cardiac drugs; this was especially true for stroke. There were strong interactions with specific cardiac drugs; for example, both BA and IB substantially increased the risk of supraventricular tachycardia in patients not anti-arryhthmic agents, but not in the presence of such agents.We conclude that bronchodilator therapy for chronic obstructive diseases is associated with increased cardiovascular risk, especially in patients without previous cardiovascular diagnoses, and that this is unlikely due to the severity of the respiratory disease, since risk was not increased with ICS.     

Cervical lymph node metastases with occult primary

Patients with cervical metastases from an unknown primary tumour present both diagnostic and therapeutic problems. Twenty patients with a metastatic lesion in the neck, in whom the primary tumour was not found despite extensive, diagnostic procedures, were treated in the Department of Otolaryngology, Link?ping University Hospital, during a 14-year period (1971-1984). Since 1975 tonsillectomy has been performed regularly on the affected side as a part of the diagnostic procedure. Eight patients without any clinical signs of a tonsillar primary have been diagnosed to have carcinoma by tonsillectomy and careful examination of the serially sectioned tonsillectomy specimen. The diagnosis of the metastatic nodes was based on histopathological examination in 8 patients and on fine needle aspiration cytology in 12. Four of 9 patients with squamous cell carcinoma survived 3 years and so did 4 of 5 with anaplastic carcinoma. All 5 patients treated surgically are alive and 6 of 12 who were irradiated. Ten of 12 patients with mobile unilateral nodes (N1) were alive at follow-up after 1-13 years but only 3 of 8 patients with fixed nodes (N3) at follow-up of 1-3 years.     

Heterogeneity of salivary gland tumors studied by flow cytometry

Intratumor DNA heterogeneity was investigated by flow cytometric analysis of multiple samples taken from different sites of 8 benign and 16 malignant primarily resected salivary gland tumors. All benign tumors had diploid DNA content. The overall incidence of DNA diploidy in 16 malignant cases examined was 50%. Intratumor differences in DNA ploidy were observed in four malignant tumors (25%); 2 of these 4 heterogenous tumors contained both aneuploid and diploid cell clones. The remaining 12 tumors showed a homogeneous DNA content in the different specimens; 8 were diploid, 3 aneuploid, and 1 was polypoid. The DNA nondiploid tumors were clinically more advanced than the DNA diploid ones (p < 0.01). The tumor proliferation rate (fraction of cells in S-phase) was higher in DNA nondiploid samples than in diploid ones (p < 0.01). The DNA nondiploid tumors seemed to recur more often than DNA diploid ones did. The data emphasize the usefulness of DNA measurements for the characterization of malignant salivary gland tumors but also the importance of adequate sampling in assessing their DNA ploidy.     

DNA analysis of malignant salivary gland carcinomas: comparison of different tissue preparations and measuring techniques.

This study correlates static and flow-cytometric analysis on paraffin-embedded and fresh material of salivary gland carcinomas. Nuclear suspensions for DNA analysis were prepared from paraffin-embedded and fresh material. Comparison of the results of static and flow cytometry on paraffin-embedded material revealed no significant correlation between DNA ploidy and S-phase value. The coefficients of variation were significantly lower for static than for flow cytometry (p < 0.05). Flow-cytometric analysis on fresh and paraffin-embedded material correlated well concerning DNA ploidy, but not for the S-phase. The coefficients of variation were significantly lower for the fresh than for the paraffin-embedded material (p < 0.001). Cytometric analysis on paraffin-embedded material of malignant salivary gland carcinomas should be critically evaluated. There is a need for flow-cytometric investigations on fresh material to clarify further the prognostic value of this method.     

Agarose gel method: its usefulness in assaying factor VIII inhibitors, evaluating treatment and suggesting a mechanism of action for factor IX concentrates

Plasmas from 14 patients with factor VIII inhibitors, 10 haemophiliacs and four non-haemophiliacs, were assayed by both the agarose gel and Bethesda methods. Good correlation was observed in 34 samples from 13 patients, but there was poor correlation in three samples from a single haemophilic patient. The sensitivity of the method was increased by diluting normal platelet rich plasma (PRP) with congenital factor VIII deficient plasma. With this modification, as little as 0.4 of a Bethesda unit (Bu) could be measured accurately. The agarose method is easier to perform and requires much less technician time than the Bethesda assay (Ba). Inhibitory activity can be measured even in the presence of large amounts of transfused factor VIII or factor IX concentrates. To study the effect of factor IX concentrates on the inhibitor, the method was modified by incorporating into the gels plasmas specifically deficient in either factor VII, IX or X. Our data suggest that factor VII is probably responsible for the 'bypassing' activity of factor IX concentrates.     

CNS myelination requires cytoplasmic dynein function

Background: Cytoplasmic dynein provides the main motor force for minus‐end‐directed transport of cargo on microtubules. Within the vertebrate central nervous system (CNS), proliferation, neuronal migration, and retrograde axon transport are among the cellular functions known to require dynein. Accordingly, mutations of DYNC1H1, which encodes the heavy chain subunit of cytoplasmic dynein, have been linked to developmental brain malformations and axonal pathologies. Oligodendrocytes, the myelinating glial cell type of the CNS, migrate from their origins to their target axons and subsequently extend multiple long processes that ensheath axons with specialized insulating membrane. These processes are filled with microtubules, which facilitate molecular transport of myelin components. However, whether oligodendrocytes require cytoplasmic dynein to ensheath axons with myelin is not known. Results: We identified a mutation of zebrafish dync1h1 in a forward genetic screen that caused a deficit of oligodendrocytes. Using in vivo imaging and gene expression analyses, we additionally found evidence that dync1h1 promotes axon ensheathment and myelin gene expression. Conclusions: In addition to its well known roles in axon transport and neuronal migration, cytoplasmic dynein contributes to neural development by promoting myelination. Developmental Dynamics 244:134–145, 2015. © 2014 Wiley Periodicals, Inc.     

Systematic review of long-term weight loss studies in obese adults: clinical significance and applicability to clinical practice

BACKGROUND: Obesity is a common health problem that requires a long-term care approach. We systematically reviewed long-term (> or =2 y) studies investigating dietary/lifestyle, pharmacologic, and surgical weight loss methods to assess (1) weight loss efficacy, defined by absolute weight loss and the proportion of subjects with > or =5% weight loss, (2) effects of weight loss on cardiovascular risk factors, and (3) applicability of findings from studies to everyday clinical practice. METHODS: The MEDLINE, HealthSTAR, and the Cochrane Controlled Trials databases were searched for studies investigating the long-term efficacy of weight loss methods in overweight and obese adults. Data were extracted for (i) weight loss after 1 y (pharmacologic studies only), 2 y, 3 y, and 4 y, (ii) proportion of subjects with > or =5% weight loss at the end of follow-up, and (iii) changes (end-of follow-up minus baseline values) in blood lipids, fasting blood glucose, and systolic and diastolic blood pressure. RESULTS: Dietary/lifestyle therapy provides <5 kg weight loss after 2-4 y, pharmacologic therapy provides 5-10 kg weight loss after 1-2 y, and surgical therapy provides 25-75 kg weight loss after 2-4 y. Weight loss of > or =5% baseline weight is not consistently associated with improvements in cardiovascular risk factors and these benefits appear to be intervention specific and occur mainly in people with concomitant cardiovascular risk factors. Weight loss studies have methodologic limitations that restrict the applicability of findings to unselected obese people assessed in everyday clinical practice. These limitations include an inadequate study duration, large proportions of subjects lost to follow-up, a lack of an appropriate usual care group, and a lack of reporting of outcomes in high-risk subgroups. CONCLUSIONS: Dietary/lifestyle and pharmacologic weight loss interventions provide modest weight loss, and may improve markers of cardiovascular risk factors although these benefits occur mainly in patients with cardiovascular risks. Studies investigating weight loss have methodologic limitations that restrict the applicability of findings to obese patients assessed in clinical practice.     

Adenoid cystic carcinoma: Significance of DNA ploidy

Background. DNA ploidy pattern is sometimes used as a prognostic factor. Heterogeneity of a tumor could, however, give false information when a single analysis is performed. Methods. Twenty-eight patients with adenoid cystic carcinomas were retrospectively studied with regard to clinicohistologic parameters, and in 24 of these the DNA pattern was assessed using flow cytometry, with multiple analysis from different tumor parts, to determine prognostic factors. Results. Of the carcinomas, 33% (8/24) were DNA aneuploid, and 17% (4/24) of the tumors showed intratumoral heterogeneity of DNA content; two of them with mixture of diploid and aneuploid stemlines. The DNA aneuploid tumors were clinically more advanced and demonstrated a higher frequency of solid architecture than did diploid tumors (p < 0.05). The S-phase values were significantly higher in aneuploid samples than in diploid ones (p < 0.05). The recurrence rate was significantly higher in patients with aneuploid tumors (75%) than with diploid ones (19%) (p < 0.05). The cumulative survival was worse for patients with aneuploid tumors than for those with diploid ones (p < 0.05). Conclusions. Our findings suggest a potentially important role for flow cytometry in evaluation of adenoid cystic carcinoma. It is of interest to observe that in some tumors both diploid and aneuploid stemlines can co-exist. If one sample is analyzed and demonstrates diploid cells, there is a 3% chance that the tumor is also heterogenous with aneuploid stemlines. If one sample demonstrates aneuploid cells, there is a 7% chance for heterogeneity with diploid cells, as well. Two samples from different tumor parts can be considered representative. © 1995 Jons Wiley & Sons, Inc.