Screening and diagnostic breast MRI: how do they impact surgical treatment? Insights from the MIPA study.

OBJECTIVES: To report mastectomy and reoperation rates in women who had breast MRI for screening (S-MRI subgroup) or diagnostic (D-MRI subgroup) purposes, using multivariable analysis for investigating the role of MRI referral/nonreferral and other covariates in driving surgical outcomes. METHODS: The MIPA observational study enrolled women aged 18-80 years with newly diagnosed breast cancer destined to have surgery as the primary treatment, in 27 centres worldwide. Mastectomy and reoperation rates were compared using non-parametric tests and multivariable analysis. RESULTS: A total of 5828 patients entered analysis, 2763 (47.4%) did not undergo MRI (noMRI subgroup) and 3065 underwent MRI (52.6%); of the latter, 2441/3065 (79.7%) underwent MRI with preoperative intent (P-MRI subgroup), 510/3065 (16.6%) D-MRI, and 114/3065 S-MRI (3.7%). The reoperation rate was 10.5% for S-MRI, 8.2% for D-MRI, and 8.5% for P-MRI, while it was 11.7% for noMRI (p ≤ 0.023 for comparisons with D-MRI and P-MRI). The overall mastectomy rate (first-line mastectomy plus conversions from conserving surgery to mastectomy) was 39.5% for S-MRI, 36.2% for P-MRI, 24.1% for D-MRI, and 18.0% for noMRI. At multivariable analysis, using noMRI as reference, the odds ratios for overall mastectomy were 2.4 (p < 0.001) for S-MRI, 1.0 (p = 0.957) for D-MRI, and 1.9 (p < 0.001) for P-MRI. CONCLUSIONS: Patients from the D-MRI subgroup had the lowest overall mastectomy rate (24.1%) among MRI subgroups and the lowest reoperation rate (8.2%) together with P-MRI (8.5%). This analysis offers an insight into how the initial indication for MRI affects the subsequent surgical treatment of breast cancer. KEY POINTS: • Of 3065 breast MRI examinations, 79.7% were performed with preoperative intent (P-MRI), 16.6% were diagnostic (D-MRI), and 3.7% were screening (S-MRI) examinations. • The D-MRI subgroup had the lowest mastectomy rate (24.1%) among MRI subgroups and the lowest reoperation rate (8.2%) together with P-MRI (8.5%). • The S-MRI subgroup had the highest mastectomy rate (39.5%) which aligns with higher-than-average risk in this subgroup, with a reoperation rate (10.5%) not significantly different to that of all other subgroups.

Magnetic resonance imaging before breast cancer surgery: results of an observational multicenter international prospective analysis (MIPA).

OBJECTIVES: Preoperative breast magnetic resonance imaging (MRI) can inform surgical planning but might cause overtreatment by increasing the mastectomy rate. The Multicenter International Prospective Analysis (MIPA) study investigated this controversial issue. METHODS: This observational study enrolled women aged 18-80 years with biopsy-proven breast cancer, who underwent MRI in addition to conventional imaging (mammography and/or breast ultrasonography) or conventional imaging alone before surgery as routine practice at 27 centers. Exclusion criteria included planned neoadjuvant therapy, pregnancy, personal history of any cancer, and distant metastases. RESULTS: Of 5896 analyzed patients, 2763 (46.9%) had conventional imaging only (noMRI group), and 3133 (53.1%) underwent MRI that was performed for diagnosis, screening, or unknown purposes in 692/3133 women (22.1%), with preoperative intent in 2441/3133 women (77.9%, MRI group). Patients in the MRI group were younger, had denser breasts, more cancers ≥ 20 mm, and a higher rate of invasive lobular histology than patients who underwent conventional imaging alone (p < 0.001 for all comparisons). Mastectomy was planned based on conventional imaging in 22.4% (MRI group) versus 14.4% (noMRI group) (p < 0.001). The additional planned mastectomy rate in the MRI group was 11.3%. The overall performed first- plus second-line mastectomy rate was 36.3% (MRI group) versus 18.0% (noMRI group) (p < 0.001). In women receiving conserving surgery, MRI group had a significantly lower reoperation rate (8.5% versus 11.7%, p < 0.001). CONCLUSIONS: Clinicians requested breast MRI for women with a higher a priori probability of receiving mastectomy. MRI was associated with 11.3% more mastectomies, and with 3.2% fewer reoperations in the breast conservation subgroup. KEY POINTS: • In 19% of patients of the MIPA study, breast MRI was performed for screening or diagnostic purposes. • The current patient selection to preoperative breast MRI implies an 11% increase in mastectomies, counterbalanced by a 3% reduction of the reoperation rate. • Data from the MIPA study can support discussion in tumor boards when preoperative MRI is under consideration and should be shared with patients to achieve informed decision-making.

Health-related Quality of Life for Children With Leukemia: Child and Parental Perceptions.

BACKGROUND: The importance of health-related quality of life (HRQoL) in patients with acute lymphoblastic leukemia (ALL) has increased in recent years. This study aimed to assess HRQoL in children with ALL, affecting factors, and the relationship between parent proxy-report and child self-report HRQoL. MATERIALS AND METHODS: A total of 59 children and their parents (both mother and father) were enrolled in this cross-sectional study. Turkish version of the Pediatric Quality of Life Inventory (PedsQL) 3.0 Cancer Modules were used to determine HRQoL. RESULTS: According to subscales of the self-report form, nausea and operational anxiety scores differed significantly by the treatment status; communication score varied considerably by the hospitalization length of stay; pain and hurt, cognitive problems, and perceived physical appearance scores differed significantly by the maternal chronic disease status (P<0.05). The presence of maternal chronic disease was significantly related to the total score of the parent-proxy report (mother) (P<0.05). There was a moderate correlation between total scores of child and mother (P<0.05, r=0.419) but not with the father. CONCLUSION: Children on-treatment had significant problems in nausea and procedural anxiety subscales; however, children who were hospitalized more had fewer issues in the communication subscale. Also, children whose mother had chronic disease had poorer HRQoL regarding pain and hurt cognitive problems and treatment anxiety. Given the importance of assessment and monitoring HRQoL in children with ALL, health professionals should be aware of how parents' chronic disease affects HRQoL. Psychosocial support should be provided to children and their parents, especially for those whose parents have a chronic illness.

Solving the preoperative breast MRI conundrum: design and protocol of the MIPA study.

Despite its high diagnostic performance, the use of breast MRI in the preoperative setting is controversial. It has the potential for personalized surgical management in breast cancer patients, but two of three randomized controlled trials did not show results in favor of its introduction for assessing the disease extent before surgery. Meta-analyses showed a higher mastectomy rate in women undergoing preoperative MRI compared to those who do not. Nevertheless, preoperative breast MRI is increasingly used and a survey from the American Society of Breast Surgeons showed that 41% of respondents ask for it in daily practice. In this context, a large-scale observational multicenter international prospective analysis (MIPA study) was proposed under the guidance of the European Network for the Assessment of Imaging in Medicine (EuroAIM). The aims were (1) to prospectively and systematically collect data on consecutive women with a newly diagnosed breast cancer, not candidates for neoadjuvant therapy, who are offered or not offered breast MRI before surgery according to local practice; (2) to compare these two groups in terms of surgical and clinical endpoints, adjusting for covariates. The underlying hypotheses are that MRI does not cause additional mastectomies compared to conventional imaging, while reducing the reoperation rate in all or in subgroups of patients. Ninety-six centers applied to a web-based call; 36 were initially selected based on volume and quality standards; 27 were active for enrollment. On November 2018, the target of 7000 enrolled patients was reached. The MIPA study is presently at the analytic phase. Key Points • Breast MRI has a high diagnostic performance but its utility in the preoperative setting is controversial. • A large-scale observational multicenter prospective study was launched to compare women receiving with those not receiving preoperative MRI. • Twenty-seven centers enrolled more than 7000 patients. The study is presently at the analytic phase.

ARE THYROID NODULES AN OBSTACLE TO MINIMAL INVASIVE PARATHYROID SURGERY? A SINGLE-CENTER STUDY FROM AN ENDEMIC GOITER REGION.

CONTEXT: Minimally invasive parathyroidectomy (MIP) procedure has become a widely accepted alternative to the standard four-gland exploration nowadays. OBJECTIVE: The aim of this study was to evaluate patients with primary hyperparathyroidism (PHPT), who had been treated with thyroidectomy and bilateral neck exploration (BNE), rather than MIP alone, due to coexisting thyroid nodules and to determine the benefits of simultaneous thyroidectomy and the possible negative outcomes of not performing this additional procedure. DESIGN: There were 185 patients who were operated for PHPT at our clinic from January 2014 to November 2016. SUBJECTS AND METHODS: 50 patients meet inclusion criteria: have thyroidectomy at the same time of parathyroid surgery, have concordant findings of parathyroid adenoma localization at preoperative MIBI-SPECT and the cervical US and have not had malignancy on fine needle aspiration biopsy (FNAB). RESULTS: The mean age of the patients was 55.3±10.4, and female to male ratio was 7:1. All patients had parathyroidectomy with BNE and thyroidectomy: 11 (22%) patients had micropapillary thyroid cancer (mPTC), 2 (4%) had papillary thyroid cancer (PTC). CONCLUSION: The results were inconclusive in clearly demonstrating which patients presenting with coexisted thyroid nodules should undergo thyroidectomy, rather than MIP, and which should be monitored for thyroid nodules after MIP. However, we consider that in cases who are not clearly indicated for thyroidectomy, MIP followed by monitoring of thyroid nodules can be the treatment approach.

IS THERE A RELATIONSHIP BETWEEN VITAMIN D STATUS AND HYPOCALCEMIA AFTER TOTAL THYROIDECTOMY?

CONTEXT: Vitamin D plays a crucial role in calcium metabolism through parathormone-dependent process. Deficiency of this important nutrient may be associated with hypocalcemia after thyroidectomy. OBJECTIVE: To evaluate the role of vitamin D in predicting hypocalcemia following total thyroidectomy. SUBJECTS AND METHODS: One hundred and fifty patients who underwent total thyroidectomy for benign or malignant thyroid disease were included in this prospective study. The association between preoperative vitamin D status and the development of hypocalcemia were investigated. RESULTS: Biochemical and symptomatic hypocalcemia were found in 28 (18.7%) and 22 (14.7%) patients, respectively. Preoperative vit D level was found significantly lower in patients with biochemical (p = 0.006) and clinical (p < 0.001) hypocalcemia in comparison to normocalcemic patients. The patients who had <10 ng/mL vit D level (severe deficiency) developed significantly more biochemical and clinical hypocalcemia than the patients with serum vit D level higher than 10 ng/mL (p = 0.030 and p < 0.001, respectively). CONCLUSIONS: Although postthyroidectomy hypocalcemia is multifactorial, vit D deficiency, particularly severe form, is significantly associated with the development of biochemical and clinical hypocalcemia. Vit D supplementation can prevent this unwanted complication in such patients.

Tactile processing in children and adolescents with obsessive-compulsive disorder.

Many obsessive-compulsive disorder (OCD) patients experience sensory phenomena, such as bodily sensations and "just-right" perceptions accompanying compulsions. We studied tactile processing in OCD by psychophysical experiments targeting the somatosensory cortex. Thirty-two children and adolescents with OCD (8 tic-related, 19 with sensory phenomena (SP)) and their sex- and age-matched controls participated in the study. After clinical assessments, two questionnaires were completed for sensory problems (Sensory Profile and Touch Inventory for Elementary-School-Aged Children). The psychophysical experiments consisted of five tasks: simple reaction time, choice reaction time, dynamic (detection) threshold, amplitude discrimination, and amplitude discrimination with single-site adaptation. The tactile stimuli were sinusoidal mechanical vibrations (frequency: 25 Hz) applied on the fingertips. Just-noticeable differences (JNDs) were found in amplitude discrimination tasks. There was no difference between the OCD group and controls in detection thresholds. However, the OCD group (especially young males) had worse amplitude discrimination (i.e., higher JNDs) than controls. Young OCD participants had reduced adaptation than young controls. Tic-related OCD participants and those with SP had higher detection thresholds than those without. Additionally, the OCD group reported more problems than controls in the Emotional/Social subset of the Sensory Profile questionnaire. The discrimination results show altered tactile processing in OCD at suprathreshold levels. This can be explained by a scaling factor modifying the sensory signal with decreasing slope at higher input levels to achieve normal Weber fractions internally. Quadratic discriminant analysis gave the best positive (76%) and negative (60%) predictive values for classifying individuals (into "OCD" or "control" groups) based on psychophysical data alone.

Effects of bentazone on lipid peroxidation and antioxidant systems in human erythrocytes in vitro.

Bentazone, a benzothiadiazole herbicide, is widely used for a variety of crops including cereals, maize, peas, rice and soy beans. The concern for human health is stil very high because bentazone is continuously monitored in environment and several studies to evaluate its potential carcinogenic effects when chronic and high doses were administered to animals. We aimed to investigate the possible effects of bentazone on lipid peroxidation, levels of glutathione and activities of antioxidant enzymes in human erythrocytes in vitro. For that, erythrocyte were incubated with bentazone in different concentrations (0-50 nM) at 37 °C for 1 hr. Bentazone showed significant increase in the levels of malondialdehyde (MDA) at the highest concentration in erythrocytes as an index of lipid peroxidation. Besides, alterations in the levels of reduced glutathione (GSH) and activities of glutathione peroxidase (GSH-Px) were observed while the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GSH-Rd) were unchanged. In conclusion, findings from this study indicate that in vitro toxicity of bentazone may be associated with oxidative stress and this work warrants further in vivo investigations.

A radiographic assessment of the prevalence of pulp stones in a group of Turkish dental patients.

AIM: To determine the prevalence of pulp stones in a group of patients using radiographs, and to assess any associations with age, gender, systemic diseases, tooth type, jaw, caries, restorations, impacted third molars and dental anomalies including dens invaginatus, taurodontism, dens evaginatus and microdontia. METHODOLOGY: A total of 519 patients were selected randomly amongst patients referred to Oral Diagnosis and Radiology Department, Faculty of Dentistry, Ankara University. After clinical inspection, full-mouth periapical radiographs were obtained. An oral radiologist examined the radiographs to identify pulp stones. Frequency distribution of teeth with pulp stones was calculated and statistical analysis was performed with chi-square test. RESULTS: Of the patients, 313 (60%) were female and 206 (40%) were male. Sixty (12%) had one or more teeth that contained pulp stones. Amongst the 13,474 teeth examined radiographically, 627 (5%) had pulp stones. There was no significant association between pulp stone occurrence and gender or systemic diseases. As age increased, the prevalence of pulp stones increased (P < 0.01). Molars had statistically more pulp stones than premolars and incisors (P < 0.001). Frequencies in both maxillary and mandibular jaw were similar. There was no association between pulp stones and impacted third molars, caries, restorations or dental anomalies. CONCLUSIONS: The prevalence of pulp stones increased with age and was significantly higher in molar teeth than premolars and incisors. There was no significant association between pulp stones and gender, systemic diseases, impacted third molars, condition of the crown and dental anomalies.

[The validity and reliability of the Turkish Pediatric Quality of Life Inventory for children 13-18 years old]. / Cocuklar Için Yasam Kalitesi Olçeginin 13-18 Yas Ergen Formunun Geçerlik ve Güvenilirligi.

OBJECTIVE: The Pediatric Quality of Life Inventory (PedsQL) is a modular instrument that measures health related quality of life, and investigates the physical and psychosocial functioning, unrelated to health, of children 2-18 years old. In this study, the objective was to evaluate the validity and reliability of the Turkish PedsQL in adolescents 13-18 years old. METHOD: The study included 230 adolescents and 230 parents. The subjects were separated into 3 groups: 1. Healthy; 2. Has an acute disease; 3. Has a chronic disease. The reliability and the validity of PedsQL were computed. RESULTS: The internal consistency of the scale (Cronbach's alpha coefficient) was 0.82 for the child self-report and 0.87 for the parent proxy report. The PedsQL scores of both the child self-report and parent proxy report showed a statistically significant low to medium level of inversely proportional correlation with many indicators of morbidity and illness burden. There was a statistically significant and direct proportional correlation between the child self-report and parent proxy report scores. CONCLUSION: The PedsQL is valid and reliable for evaluating the quality of life of Turkish adolescents 13-18 years old.

Concomitant administration of uracil-tegafur and leucovorin during adjuvant radiotherapy for locally advanced rectal cancer.

PURPOSE: We report the feasibility and toxicity profile, and the impact on local control, disease-free survival and overall survival rates of our study which consisted of postoperative concurrent chemoradiotherapy, followed by adjuvant chemotherapy using uracil-tegafur (UFT)/leukovorin (LV) in locally advanced rectal cancer patients. PATIENTS AND METHODS: Thirty-one patients operated for rectal adenocarcinoma (pT3/4 or N+) were enrolled onto the study. Twenty-three patients were males and 8 females with median age 62 years (range 21-85). Radiotherapy (RT) to the pelvis with conformal technique and individual blocks was delivered within 8 weeks following surgery. Total RT dose was 50.4 Gy and was given in a conventional single fraction of 1.8 Gy per day. Chemotherapy was administered concomitantly and consisted of UFT (300 mg/m(2)/day) and LV (30 mg/day) during RT-days. Following chemoradiotherapy, chemotherapy alone was administered for 4 cycles in the same dose for 28 days every 35 days. RESULTS: No lethal toxicity occurred. All patients completed the scheduled RT. Concurrent chemotherapy continued in 22 (70.9%) patients until the end of RT. Seventeen (54.8%) patients completed the whole concurrent chemoradiotherapy and adjuvant chemotherapy as planned. No grade 3-4 stomatitis/mucositis or haematological toxicities were observed during the whole treatment period. During concomitant therapy grade 1-2 toxicities were: nausea/vomiting 60%, dyspepsia/gastric pain 39%, diarrhea 39% and dysuria 10%, whereas grade 3 nausea and diarrhea occurred in 6% and 19%, respectively. Median follow-up was 22 months. Two-year local control, disease-free survival and overall survival rates were 96.3%, 72.3% and 83.2%, respectively. CONCLUSION: The acute toxicity profile of UFT/LV, local control, disease-free survival and overall survival in the concurrent chemoradiotherapy setting for operated, locally advanced rectal cancer seem comparable with the standard 5-fluorouracil (5-FU)-based therapies.

Substance Abuse Profiles of Patients Admitted to the Alcohol and Drug Addiction Research, Treatment, and Education Center in Turkey.

OBJECTIVES: To determine the substance abuse profiles of patients treated a Drug Addiction Research, Treatment, and Education Center (AMATEM) in association with the percentage of substance use distribution and multiple substance use in their urine samples. For this, we retrospectively evaluated the urine sample analysis reports of 600 male and female patients aged 13 to 65 years who were treated at the AMATEM unit of Istanbul Neuropsychiatry Hospital between January 1st, 2015, and December 12th, 2015. MATERIALS AND METHODS: The urine samples were sent to Üsküdar University Advanced Toxicology Analysis Laboratory and were analyzed using a UPLC tandem mass spectrometer (UPLC-MS/MS). To determine the substance use profiles of the patients applying to AMATEM, statistical assessment was performed on the analysis reports of the patients. RESULTS: When the analysis reports of the 600 urine samples were examined, 293 patients were identified to have used addictive substances. The substances most frequently detected in the urine samples were respectively: cannabis, alcohol, morphine, cocaine, synthetic cannabinoids, 3,4-Methylenedioxymethamphetamine, and amphetamine. CONCLUSION: The findings in our study resemble the rates of cannabis use by the young population throughout the world. Our results show differences to the literature regarding the consumption of synthetic cannabinoids because the variety of synthetic cannabinoids change rapidly around the world each year.

Protective effects of antithrombin III on retinal ischemia/reperfusion injury in rats: a histopathologic study.

PURPOSE: To investigate the effect of antithrombin III (AT III) on retinal ischemia/reperfusion (I/R) injury in rats. METHODS: The study was carried out on 10 Wistar albino rats (20 eyes) and four-vessel occlusion method was employed to induce retinal ischemia in this study. Rats were divided into two groups: Group I (control group, 10 eyes) and Group II (AT III, 10 eyes). In both groups, vertebral arteries were occluded bilaterally an electric needle coagulator under an operating microscope. A total of 48 hours after the initial procedure, the rats were re-anesthetized and both common carotid arteries were clamped to interrupt blood flow. In Group II, rats were injected intravenously with 250 U/kg of AT III 5 minutes before the induction of ischemia. Duration of ischemia was 30 minutes. At the end of this period, clamp was removed for the reperfusion of the eye for 4 hours. Following the reperfusion period, the animals were killed by decapitation. Retinal sections were evaluated under light and electron microscope. The signs of I/R injury at the microscopic level, i.e., cellular degeneration, vacuolization between retinal layers, increase in the retinal thickness due to edema, mononuclear cell infiltration, and apoptotic cells, were recorded for each group. RESULTS: Retinal sections obtained from the rats in the AT III group revealed a well preserved retinal structure. When average thickness values of the two groups were compared to each other, the difference was significant with respect to inner nuclear and inner plexiform layers indicating increased retinal thickness values in Group I due to tissue edema resulting from I/R injury. Similarly, mononuclear cell infiltration and apoptotic cell counts were found to be significantly higher in control group compared to AT III group showing the inhibitory effect of AT III on leukocyte infiltration and apoptotic cell death in rat retina. CONCLUSIONS: Antithrombin III attenuated I/R injury in rat retina.

Serum irisin levels in patients with polycystic ovary syndrome.

OBJECTIVE: Polycystic ovary syndrome (PCOS) is clinically heterogeneous endocrine disorders. Insulin resistance-related proteins play a role in the etiopathogenesis of PCOS. Irisin is a newly identified myokine which act like adipokines. Irisin has been shown to be associated with the insulin resistance and metabolic syndrome. The purpose of this study was to determine the serum levels of irisin in PCOS patients and evaluate the correlations with other metabolic and hormonal parameters. PATIENTS AND METHODS: Thirty-five PCOS patients and 35 matched healthy controls were enrolled to study. Serum irisin levels, anthropometric, hormonal and metabolic parameters including HOMA-IR were measured. Linear regression analysis was employed to study the relationship between irisin and hormonal and metabolic parameters. RESULTS: Serum irisin level in PCOS patients (mean value; 0.491±0.145 µg/mL) was significantly elevated when compared to control group (mean value 0.281±0.138 µg/mL) (p < 0.001). Linear regression analysis showed that serum irisin was positively associated with body mass index, luteinizing hormone, fasting insulin and total cholesterol in the overall patient population but not for PCOS group alone (p < 0.05). CONCLUSIONS: Serum irisin level of PCOS patients was high compared to that of healthy control subjects. In patients with PCOS, this situation may be due to insulin resistance, when there is leptin resistance or metabolic syndrome.

Functional analysis of two long terminal repeats from the HTLV-I retrovirus.

Human T-cell leukemia virus type I (HTLV-I) is associated with a large spectrum of clinical manifestations in man. Viral and host factors are probably involved in determining the consequences of infection. Although most of the genome of HTLV-I appears remarkably stable, considerable variation is observed in the long terminal repeat (LTR) which harbors the promoter region. So far, no correlation between specific mutations and pathogenesis has been found, and the current opinion is that sequence variations reflect the geographical origin of the isolate more than the associated pathology. To assess whether the mutations observed between two HTLV-I LTRs were functionally significant, two LTRs, which differ by ten mutations, were coupled to the highly sensitive eukaryotic luciferase-encoding reporter gene, luc, and tested by transfection in a variety of cell lines. Marked differences in promoter activity were observed in some of the cells tested, whereas in other both LTRs were equally active. This result demonstrates that the minor differences observed between two HTLV-I LTRs can affect the activity level of the promoter in some cellular environments, a result which could point to the LTR as one determinant of HTLV-I cell tropism in vivo.

Comparison of the sensitivities of ribonucleic acid and oligonucleotide probes for in situ detection of Theiler's virus mRNA.

To evaluate the sensitivity of in situ detection of the Theiler's virus genome, we hybridized BHK-21-infected cells with antisense ribo- and oligonucleotide 35S-labeled probes. The sensitivity achieved with the anti-sense 280-nucleotide riboprobe was similar to that obtained with a 93-mer oligonucleotide probe. However, more reproducible and accurate results were obtained with the riboprobe. With long exposure times, the background was higher with the oligonucleotide probe than with the RNA probe. The background was improved by using freshly labeled oligonucleotide probe.

In situ analysis of proteolipid protein gene transcripts during persistent Theiler's virus infection.

SJL/J mice inoculated intracranially with the DA strain of Theiler's virus exhibit a persistent demyelinating disease of the central nervous system. To investigate the effect of persistent infection of oligodendrocytes on the expression of myelin genes, we analyzed the level of PLP mRNA in infected as well as uninfected oligodendrocytes. This study was performed at the single-cell level using the simultaneous detection of viral antigens by immunocytochemistry and PLP mRNAs by in situ hybridization with 35S-labeled oligonucleotide probes. Our data indicate that viral infection of oligodendrocytes reduces the level of PLP mRNA by about 80%.

Simultaneous in situ detection of two mRNAs in the same cell using riboprobes labeled with biotin and 35S.

We used 35S-labeled and biotinylated cRNAs (riboprobes) to detect simultaneously two different mRNAs by in situ hybridization. In a first step we established the conditions under which each type of probe achieved the same high level of sensitivity. We then used these conditions to hybridize BHK cells infected with Theiler's virus, a murine picornavirus, with a mixture of a virus-specific biotinylated riboprobe and a 35S-labeled riboprobe specific for beta-actin mRNA. Both mRNAs could be detected in the same cell, although the sensitivity achieved by the radiolabeled probe was reduced by about 40% by the simultaneous hybridization with the biotinylated probe.

Comparative analysis of HTLV-I promoter activities reveals no disease-linked pattern of expression.

To investigate the possibility of an association between the type of pathology caused by HTLV-I and the activity of its promoter, we compared the levels of transcription obtained with six LTRs isolated from patients with two different HTLV-I-related diseases: ATL and TSP/HAM. The patients came from different geographical endemic areas. The LTR region was amplified by polymerase chain reaction (PCR) from the DNA of uncultured peripheral blood lymphocytes, and directly cloned upstream of the luciferase reporter gene. Constructs were tested by a transient transfection assay in a variety of cell lines. Although the activities of these LTRs were statistically different in some of the cell lines tested, no correlation could be demonstrated between the promoter activity and the nature of the disease. Thus, the data suggest that the LTR is not a major determinant of the nature of the disease associated with the infection by HTLV-I.

Acylated delphinidin glycosides in the blue-violet flowers of Consolida armeniaca.

Four new acylated delphinidin 3,7-glycosides were isolated from the blue-violet flowers of Consolida armeniaca as major anthocyanin pigments. The first pigment was based on delphinidin 3,7-diglucoside and was identified as delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7-O- [6-O-(4-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)-p-hydroxybenzoy l)- beta-D-glucopyranoside] by spectral methods. The other three acylated anthocyanins were based on delphinidin 3-glucoside-7-sophoroside. The second pigment was determined to be delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7-O-[2-O-(beta-D- glucopyranosyl)-6-O-(4-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)- p- hydroxybenzoyl)-beta-D-glucopyranoside]. The third pigment was elucidated to be delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7-O-[2-O-(6-O-(p- hydroxybenzoyl)-beta-D-glucopyranosyl)-6-O-(4-O-(6-O-(p-hydroxybenzoyl)- beta-D- glucopyranosyl)-p-hydroxybenzoyl)-beta-D-glucopyranoside], and the fourth as delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7- O-glucopyranosyl[(2-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)-6-O - (4-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)-p-hydroxybenzoyl)-be ta-D- glucopyranoside].