[Fascia iliaca Block with General Anaesthesia vs. Subarachnoid Morphine Block in Total Hip Arthroplasty]. / Blokáda fascia iliaca s celkovou anestezií vs. subarachnoidální blokáda s morfinem u totální náhrady kycelního kloubu.

PURPOSE OF THE STUDY Many physicians believe that loco-regional anaesthesia and analgesia improve the postoperative course of patients indicated for total hip arthroplasty compared to general anaesthesia. However, there are many patients who refuse subarachnoid or epidural anaesthesia, or have contraindications or conditions making the use of such techniques impossible. An alternative option is the combination of general anaesthesia and a peripheral nerve blockade. The aim of this prospective randomized open-label clinical trial was to compare the efficacy and quality of postoperative analgesia between fascia iliaca block combined with general anaesthesia (GA) and subarachnoid anaesthesia with morphine and bupivacaine (SAB). MATERIAL AND METHODS After having obtained the ethics committee approval and the patients consent, a prospective, open-label, randomized trial was conducted in patients referred for total hip arthroplasty (THR). The GA group was administered ultrasound-guided fascia iliaca block with 40 ml of 0.25% bupivacaine solution after the induction of general anaesthesia. In the SAB group, subarachnoid blockade was performed with a mixture of 3 ml of 0.5% bupivacaine with 0.150 mg morphine prepared in the hospital pharmacy. Right after surgery the patients were taken to the ICU for 24 hours, after which they were transferred to a general ward. In addition to vital signs monitoring, pain intensity using a 0-10 numeric rating scale (NRS), first morphine administration at NRS >4, total morphine consumption and potential adverse effects were observed over the period of 72 hours. RESULTS There was no statistical difference between the GA (14 persons) and the SAB (14 persons) group in demographic parameters, time to first morphine administration (10 hrs vs. 19 hrs, p=0.10), number of persons with no need for morphine after surgery (5 vs. 7), tingling sensation (1 vs. 0) or numbness of the limb (0 vs. 1). There was no difference in cardiorespiratory parameters or side effects of therapy. In neither case was there respiratory depression or delayed rehabilitation. No patient developed delirium after surgery, and no patient reported dissatisfaction with pain management. DISCUSSION The fascia iliaca block and subarachnoid anaesthesia using local anaesthetic with opioid addition have been repeatedly published for patients after total hip arthroplasty, but this study is unique by comparing the two methods. The study added a new piece of knowledge to the findings of several recent meta-analyses on the comparable outcomes of general and subarachnoid anaesthesia for hip replacement in the perioperative period. CONCLUSIONS If subarachnoid anaesthesia cannot be used in hip arthroplasty, general anaesthesia with fascia iliaca block provides comparable analgesia and quality of postoperative course. Key words: total hip arthroplasty, general anaesthesia, fascia iliaca block, subarachnoid anaesthesia, postoperative analgesia, postoperative course.

Can changes in skin impedance be used to monitor sedation after midazolam and during recovery from anesthesia?

It has been suggested that sympathetic activity, measured as changes in electrical skin impedance (SI), can be used to assess the adequacy of general anesthesia. Our prospective study investigated if measurements of skin impedance can determine levels of sedation induced by midazolam. Twenty-seven patients scheduled for arthroscopy requiring general anesthesia were served as their own control. These were blinded to the order of injections by telling them that they will be randomly administered a placebo (saline) orsedative agent. A DM 3900 multimeter was used for SI measurements. The degree of sedation was measured using the modified Observer's Assessment of Alertness and Sedation (mOAAS) scale. Resting SI values were noted, and all participants were then administered the placebo followed 5 min later by midazolam 2 mg i.v. Five min after that, patients were administered standard general anesthesia with propofol, oxygen, nitrous oxide 60 %, and isoflurane 1 MAC via a laryngeal mask, and sufentanil 5 - 10 µg. SI significantly increased after administration of midazolam and induction of anesthesia. There were no significant differences between pre-administration (baseline) and placebo and end of surgery and end of anesthesia with closed eyes. There were highly significant differences (p<0.001) between pre-administration vs. midazolam, placebo vs. midazolam, pre-administration vs. induction of anesthesia. We found slight correlation between mOAAS and SI. There were no significant changes between the end of surgery and the end of anesthesia with closed eyes, but SI significantly decreased (p<0.01) after eyes opened.

Late outcomes in survivors of childhood acute myeloid leukemia: a report from the St. Jude Lifetime Cohort Study.

Cumulative burden of chronic health conditions and neurocognitive and physical function were examined among survivors of childhood acute myeloid leukemia (AML) treated with hematopoietic cell transplant (HCT; n = 66) or conventional therapy (CT; n = 67). Survivors and controls underwent a comprehensive clinical assessment, and health conditions were graded using a modified version of the Common Terminology Criteria for Adverse Events. By age 40 years, HCT and CT survivors had an average 17.4 (95% confidence interval [CI] 14.6-20.1) and 9.3 (7.7-11.1) grade 1-4 conditions versus 3.8 (3.3-4.2) in community controls. Compared to controls, HCT survivors had a higher prevalence of hypertriglyceridemia (45.5% vs. 18.3%), hypercholesterolemia (47.0% vs. 30.9%), hypothyroidism (27.3% vs. 4.0%), and primary hypogonadism (p < 0.001). CT survivors had a higher prevalence of cardiomyopathy (11.9% vs. 2.7%) and hypertension (53.7% vs. 44.3%). Neurocognitive impairment was elevated across all domains compared to controls but did not differ by treatment modality. Compared to controls, a higher proportion of HCT survivors had impairments in strength and endurance; whereas flexibility and mobility impairments were noted among CT survivors. Despite successful advances in childhood AML therapy, many therapeutic exposures remain unchanged. These findings support ongoing investigations of novel therapies and strategies to ameliorate the risk of late morbidities.

Subsequent Breast Cancer in Female Childhood Cancer Survivors in the St Jude Lifetime Cohort Study (SJLIFE).

PURPOSE: Anthracycline-associated risk for subsequent breast cancer in childhood cancer survivors is hypothesized to be mediated by TP53 mutation-related gene-environment interactions. We characterized treatment/genetic risks and the impact of screening for breast cancer in the St Jude Lifetime Cohort. PATIENTS AND METHODS: Female participants underwent risk-based assessments, prior health event validation, chest radiation dosimetry, and whole genome sequencing. Breast biopsy reports were reviewed. A subgroup (n = 139) underwent both breast magnetic resonance imaging and mammography. Multivariable regression was used to calculate hazard ratios (HRs) and 95% CIs. RESULTS: Among 1,467 women, 56 developed 68 breast cancers at a median age 38.6 (range, 24.5 to 53.0) years. Cumulative incidences at age 35 years were 1% (no chest radiation) and 8% (≥ 10 Gy of chest radiation). In adjusted models, breast cancer was associated with 20 Gy or more of chest radiation versus none (HR, 7.6; 95% CI, 2.9 to 20.4), anthracycline exposure versus none (1 to 249 mg/m2: HR, 2.6; 95% CI, 1.1 to 6.2; ≥ 250 mg/m2: HR, 13.4, 95% CI, 5.5 to 32.5), and having a breast cancer predisposition gene mutation (HR, 23.0; 95% CI, 7.3 to 72.2). Anthracyclines 250 mg/m2 or greater remained significantly associated with increased risk of breast cancer in models excluding survivors with cancer predisposition gene mutations, chest radiation 10 Gy or greater, or both. Sensitivity/specificity were 53.8%/96.3% for mammography, 69.2%/91.4% for magnetic resonance imaging, and 85.8%/99.7% for dual imaging. Breast cancers detected by imaging and/or prophylactic mastectomy compared with physical findings were more likely to be in situ carcinomas, smaller, without lymph node involvement, and treated without chemotherapy. CONCLUSION: Higher doses of anthracyclines are associated with increased risk of breast cancer independent of mutations in known cancer predisposition genes. Surveillance imaging identifies breast cancers less likely to require chemotherapy than those detected by physical findings.

Neurocognitive, psychosocial, and quality-of-life outcomes in adult survivors of childhood non-Hodgkin lymphoma.

BACKGROUND: Children with non-Hodgkin lymphoma (NHL) undergo treatment with central nervous system-directed therapy, the potentially neurotoxic effects of which have not been reported in NHL survivors. METHODS: NHL survivors (n = 187) participating in the St. Jude Lifetime Cohort who were 10 or more years from their diagnosis and were 18 years old or older underwent neurocognitive, emotional distress (Brief Symptom Inventory 18), and health-related quality of life (HRQOL) assessments (36-Item Short Form Health Survey). Age-adjusted z scores were compared with community controls (n = 181) and normative data. Treatment exposures were abstracted from medical records. Models adjusted for the age, sex, and time from diagnosis were used to calculate the risk of impairment. RESULTS: The mean ages at evaluation were similar for the survivors and the controls (35.7 ± 8.9 vs 35.5 ± 11.0 years; P = .86). Survivors were 25.2 ± 8.8 years from their diagnosis: 43 (23%) received cranial radiation, 70 (37%) received high-dose methotrexate, 40 (21%) received high-dose cytarabine, and 151 (81%) received intrathecal chemotherapy. Survivors' intelligence and attention were within normal limits; however, their memory, executive function, processing speed, and academics were impaired in comparison with both population norms and community controls (P values < .05). Treatment-related exposures were not associated with neurocognitive function; however, neurocognitive impairment was associated with lower educational attainment, unemployment, and occupational status (P values < .03). Slower processing speed and worse self-reported executive function were associated with symptoms of depression (P values ≤ .003) and poorer HRQOL (P values < .05). CONCLUSIONS: Adult survivors of childhood NHL experience impaired neurocognitive function, which is associated with lower social attainment and poor HRQOL. Early-detection and intervention strategies are recommended. Cancer 2017. © 2017 American Cancer Society.

Where cancer genomics should go next: a clinician's perspective.

Large-scale, genomic studies of specific tumors such as The Cancer Genome Atlas have provided a better understanding of the alterations of pathways involved in the development of solid tumors including glioblastoma, breast cancer, ovarian and endometrial cancers, colon cancer and lung squamous cell carcinoma. This tremendous effort of the scientific community has confirmed the view that cancer actually represents a wide variety of diseases originating from different organs. These studies showed that TP53 and PI3KCA are the two most mutated genes in all types of cancers and that 30-70% of all solid tumors harbor potentially 'actionable' mutations that can be exploited for patient stratification or treatment optimization. Translation of this huge oncogenomic data set to clinical application in personalized medicine programs is now the main challenge for the future. The gap between our basic knowledge and clinical application is still wide. Closing the gap will require translational personalized trials, which may initiate a radical change in our routine clinical practice in oncology.

[A combination of dexmedetomidine with ketamine and opioids results in significant inhibition of hemodynamic changes associated with laparoscopic cholecystectomy and in prolongation of postoperative analgesia]. / Kombinace dexmedetomidinu s ketaminem a opioidy významne potlacuje hemodynamické zmeny zpusobené laparoskopickou cholecystektomií a prodluzuje trváiní pooperacní analgezie.

THE AIM OF THE STUDY: Recently, alpha2 sympathoadrenergic drugs are used in premedication to improve the perioperative course. The aim of our study was to compare a premedication with a new alpha2 sympathoadrenergic drug and standard premedication. METHODS: After ethic committee approval and written patient consent, in a randomised, double-blinded study, combination of dexmedetomidine 1.0 microg x kg(-1) + ketamine 0.5 mg x kg(-1) + fentanyl 1.0 microg x kg(-1) + atropine 0.5 mg (group FNT), dexmedetomidine 1.0 microg x kg(-1) + ketamine 0.5 mg x kg(-1) + alfentanil 5.0 microg x kg(-1) + atropine 0.5 mg (group ALFNT), or pethidine 1.0 mg x kg(-1) + atropine 0.5 mg (group Dolsin) was administered to a deltoid muscle 15 min. before anaesthesia (GA) in patients elicited for laparoscopic cholecystectomy (LCHE). GA was performed in a standard way, ECG, NIBP, respiration rate, SpO2, onset of effect, Observers Assessment of Alertness Sedation Score (OAASS) before GA, circulatory reaction to intubation and capnoperitoneum, fentanyl consumption during GA, time to the first request for post-operative analgesia and postoperative nausea and vomiting were measured. The data were processed by Kruskal-Wallis and Fisher tests. P-value < 0.05 was considered significant. RESULTS: There were 16 patients in FNT and Dolsin and 15 patients in ALFNT with no differences in demography except for younger age in ALFNT. The main differences were in hypertension during capnoperitoneum: 0/16 FNT and 1/15 ALFNT vs. 11/16 Dolsin, both p < 0.001, per-operative fentanyl consumption: FNT 31.5 microg vs. Dolsin 165.0 microg, p < 0.001 and ALFNT 50.0 microg, p < 0.05 (ALFNT vs. Dolsin, p < 0.01) and request to the first analgesic post surgery: FNT 1.3 h. vs. Dolsin 0.45 h., p < 0.05 vs. ALFNT 0.8 h., p < 0.01. There were no differences in side effects except for bradycardia in ALFNT (p < 0.05). CONCLUSIONS: Dexmedetomidine-ketamine-fentanyl-atropine combination is superior to pethidine-atropine combination in suppressing of adverse hemodynamic effects of capnoperitoneum, decreased need for analgesia during GA and prolonged postoperative analgesia.

[Chronic post-mastectomy pain]. / Chronická bolest po operaci prsu.

BACKGROUND: Surgery results in chronic pain in 7-80 percent. One of the most studied is chronic post-mastectomy pain. The prevalence was 40-50 percent in studies performed abroad. As this problem has not yet been studied in the Czech Republic, a retrospective prevalence study was performed to asses the extent of the problem and risk factors for development of chronic post-mastectomy pain. METHODS AND RESULTS: After ethic committee approval an anonymous questionnaire was developed and distributed in various oncology department and patients'organisations. Response rate was 100 percent, 330 questionnaires were processed. Chronic post-mastectomy pain (lasting longer than 3 months after surgery) was described by 69 (20.9 per cent) women. The pain was permanent in 17 and transient in 46 cases, not specified in 6 cases. The pain intensity was predominantly mild or moderate. Risk factors were younger age (below 55-60 years, p=0.0098), less extensive surgery (tumourectomy vs. mastectomy, p=0.0017), intensive post operative pain (p=0.0002) and radiotherapy (p=0.0174). Trend in chemotherapy (p=0.0778) was observed. CONCLUSIONS: The prevalence of chronic post-mastectomy pain was lower in our study comparing to studies in other countries. The reason remains obscure in spite of detailed analysis.

[Patients' perioperative worries and experiences]. / Obavy a prozitky pacientu v perioperacním období.

AIM: The aim of this study was to detect the main causes of patients' preoperative distress and their subjective experiences. METHODS: After institutional ethic committee approval and patients consent an anonymous questionnaire was randomly distributed on the first and second postoperative day. Its content were basic demography, preoperative anxiety level using five point verbal scale, main cause; of anxiety, the worst postoperative experience, postoperative pain intensity (11 point scale) and its treatment incidence of postoperative nausea and vomiting (PONV) and other side effects of the surgery and the anesthesia. Correlations with specific patients' characteristics were searched for to determine the risk groups. RESULTS: Two hundred filled in questionnaires out of the 244 distributed could be used for further evaluation. Sixty-two per cent of patients suffered from preoperative anxiety, more women (p < 0.01) and more frequently after intra-abdominal (intra-thoracic) surgeries (p < 0.05). Forty-three per cent of patients had no subjective problems following surgery, others suffered more frequently from the postoperative pain (18.5 percent), PONV (11.5 percent) and postoperative procedures, e.g. changing of dressing, drains and from the postoperative regime. There was a correlation between PONV and both the intensity of postoperative pain and anamnesis of PONV during previous surgery (p < 0.01).

[Diclofenac 75mg. and 30 mg. orfenadine (Neodolpasse) versus placebo and piroxicam in postoperative analgesia after arthroscopy]. / Diklofenak 75 mg s orfenadinem 30 mg (NEODOLPASSE) versus placebo a piroxikam v pooperacní analgezii u artroskopií.

PURPOSE OF THE STUDY: Arthroscopy is often performed in an out-patient department or as one-day surgery. Opioids often used as postoperative analgesics may have unwanted side effects that may postpone the patient's discharge from hospital. This study was designed to evaluate a substitute for the most frequently used opioid pethidine. For pain relief, non-steroid anti-inflammatory drugs are recommended, but they offer a limited choice for parenteral administration. We used a new agent (Neodolpasse) based on diclophenac and orphenadine, and compared its efficacy with piroxicam and placebo. METHODS: A total of 119 patients scheduled for knee joint arthroscopy were included in this prospective study. In a randomized, double-blind manner, they received piroxicam (P), Neodolpasse (combining 75 mg diclophenac and 30 mg orphenadine; N) or placebo (C). The number of patients in groups P, N and C were 44, 35 and 40, respectively. The effect of therapy was evaluated on the basis of the following criteria: duration of post-operative analgesia until a request for another analgesic, pain intensity (0-10 VAS), side effects and the patient's satisfaction with analgesia. The efficacy was evaluated for 24 hours after arthroscopy; premedication and analgesia induction and administration followed the same anesthetic protocol in all groups. The ethic committee approved the study and patients gave their informed consent. The results were statistically evaluated using the ANOVA analysis of variance completed by a multiple comparison of levels of significance according to Bonferroni. The presence of side and unwanted effects was analyzed by the chi-square of Fisher's exact test. A p value les than 0.05 was regarded as statistically significant. RESULTS: There were significant differences in the number of patients not requiring further analgesic medication after arthroscopy (P 52.3% vs. C (11.7%) p < 0.05, N (68.6%) vs. C p < 0.001), lower average postoperative pain (0 to 10-point scale, P 2.4 vs. C 2.9 p < 0.05, N 1.5 vs. C p < 0.05) and fewer side effects (N vs. both P and C, p < 0.05). DISCUSSION: The combination of diclophenac with orphenadine for intravenous application has only recently been available in the Czech Republic. The addition of a central muscle relaxant to a peripheral analgesic has a better effect than diclophenac alone. This may also account for a longer duration of analgesia in comparison with piroxicam reported to have significantly longer analgesic effects. The new medication also had fewer side effects. It was interesting to record that even the patients who had more pain and shorter postoperative analgesia were satisfied with the therapy provided. CONCLUSIONS: The main result of this study is the finding that Neodolpasse significantly reduces the intensity of postoperative pain and increases the duration of postoperative analgesia after knee joint arthroscopy.

Identifying genes for neuron survival and axon outgrowth in Hirudo medicinalis.

We have studied the molecular basis of nervous system repair in invertebrate (Hirudo medicinalis) nerve cells. Unlike in mammals, neurons in invertebrates survive injury and regrow processes to restore the connections that they held before the damage occurred. To identify genes whose expression is regulated after injury, we have used subtractive probes, constructed from regenerating and non-regenerating ganglia from the leech Hirudo medicinalis, to screen cDNA libraries made from whole leech CNS or from identified microdissected neurons. We have identified genes of known or predicted function as well as novel genes. Known genes up-regulated within hours of injury and that are widely expressed in invertebrate and mammalian cells include thioredoxin and tubulin. Other known genes, e.g. Cysteine Rich Intestinal Protein (CRIP), have previously been identified in mammalian cells though not in regenerating adult neurons. Two regulated genes identified, myohemerythrin and the novel protein ReN3 are exclusively expressed in invertebrates. Thus our approach has enabled us to identify genes, present in a neuron of known function, that are up- and down-regulated within hours of axotomy, and that may underpin the intrinsic ability of invertebrate neurons to survive damage and initiate regrowth programmes.

[Progress in surgical treatment may improve the quality of life in patients with breast surgery for malignant tumors]. / Pokroky v chirurgické lécbe mohou zlepsovat kvalitu zivota pacientek po operaci prsu pro zhoubný nádor.

Breast cancer surgery (BCS) is a common procedure performed in women. Chronic postmastectomy pain (PMP) has been reported in as many as 22-72 percent of patients. There are no published reports in Czech Republic. This study was performed to evaluate the prevalence and risk factors of PMP. An anonymous questionnaire was given to women after attending oncology departments in various hospitals during 3-month period. All 176 questionnaires have returned. The average age was 63.6 (SD 11.2) years, BMI 26.3 (SD 4.0). There was 23.6% of PMP prevalence in our group. The risk factors were younger age (below 60, p < 0.001, below 65, p < 0.01 resp.) and type of surgery (total mastectomy vs. tumour removal, p < 0.01). The prevalence of PMP in our study was significantly lower than in other studies. The age of our group was higher, than in other studies, but it does not explain the difference. We can hypothesise the reason is improved surgical technique.

Radiation hybrid map, physical map, and low-pass genomic sequence of the canine prcd region on CFA9 and comparative mapping with the syntenic region on human chromosome 17.

Progressive rod-cone degeneration (prcd) is a canine retinal disease that maps to the centromeric end of CFA9 in a region of synteny with the distal part of HSA17q. As such, prcd has been postulated as the only animal model of RP17, a human retinitis pigmentosa locus that maps to 17q22. In an effort to establish more detailed regions of synteny between dog CFA9 and the HSA17q-ter region, we created a robust gene-enriched CFA9-RH08(3000) map with 34 gene-based markers and 12 microsatellites, with the highest resolution and number of markers for the centromeric end of CFA9. Furthermore, we built an approximately 1.5-Mb physical map containing both GRB2 and GALK1, genes so far identified by meiotic linkage analysis as being closest to the prcd locus, and generated about 1.2 Mb low-pass (3.2x) canine sequence. Canine to human comparative sequence analysis identified 49 transcripts that had been previously mapped to the HSA17q25 region. The generated low-pass canine sequence was annotated with a working draft of human sequence from HSA17q25, and we used this scaffold to order and orient the canine sequence against human. This order and orientation are preliminary, as high-throughput genomic sequencing of HSA17q-ter has not been fully completed.

New type of anaesthesia with sevofluran improves surgical and post-surgical course of burns treatment (IGA 6116/2).

The objectives of this open-perspective clinical study were to test the effect of a new type of anaesthesia using sevofluran during surgical treatment of patients with burns, and to compare it with that of ketamin, at present the most frequently used anaesthetic. The study, conceived as a pilot study, was performed on 10 paediatric and 8 adult patients with 2nd degree burns covering between 7% and 35% of body surface. Owing to highly significant differences in parameters in favour of sevofluran, it was recommended by our ethical committee not to increase the number of patients. The parameters considered were ease of induction, undesirable effects, span of time before oral administration of liquids. Daily uptake of energy was measured for both types of anaesthetics in another group of 10 patients. In adult patients the two anaesthetics, ketamin versus sevofluran, differed in terms of incidence of unrest during surgery (ketamin 50% vs. sevofluran 0%, p < 0.05), time between end of surgery and return to full consciousness (11.3 min. vs. 2.7 min. p < 0.001), post-surgical inhibition (75% vs. 0%, p < 0.01) psychomimetic reactions after surgery (50% vs. 0%, p < 0.05), time until first intake of liquids (86.7 min. vs. 8.5 min, p < 0.001), and mobilisation (110.8 min. vs. 17 min., p < 0.001). In paedriatic patients, differences in speed of onset of anaesthesia (216 sec. vs. 66 sec., p < 0.001) and time until first uptake of liquids (75 min. vs. 22 min., p < 0.001) were statistically significant. Apart from problems associated with anesthesia, another considerable advantage of the new anaesthetic sevofluran was a significantly increased uptake of calories after use than was the case after ketamin (1645 kJ higher on the average, p < 0.05). Based on these results, sevofluran was introduced to burns surgery as a routine anaesthetic.

[Infiltration of local anesthetics into the thyroid gland capsule for surgery and the postoperative period]. / Prínos infiltrace pouzdra stítné zlázy lokálním anestetikem pro operacní a bezprostrední pooperacní prubeh.

In a prospective randomized study the hypothesis was tested whether infiltration of the thyroid capsule by a local anaesthetic will reduce the haemodynamic response to surgical trauma, consumption of anaesthetics and opioids during surgery and will shorten the time of arousal. A total of 64 patients indicated for planned goitre surgery were divided at random into a control group (C, n = 32) and experimental group (LA, n = 32). The preoperative medication and anesthesia did not differ in the two groups. In group C into the thyroid capsule a maximum of 40 ml saline was administered, in LA the same volume of 0.5% trimecain. A highly significant difference was found (p < 0.001) in the incidence of hypertension during surgery (21 C vs. 5 LA) and the need of further pharmacological interventions (21 vs. 8). In the control group was a higher consumption (p < 0.95) of the opioid phentanyl (167.5 +/- 111 micrograms vs. 125 +/- 93.5 micrograms), a trend of longer arousal and the need to antagonize the effect of opioid (p < 0.1). The substitution of saline by a local anaesthetic for infiltration of the thyroid capsule is a safe and simple method leading to a reduction of cardiovascular complications during surgery.

Effect of agroclavine on NK activity in vivo under normal and stress conditions in rats.

Agroclavine is a natural, clavine type of ergot alkaloid with D1 dopamine and a-adrenoceptor agonistic properties. We showed previously that in vitro agroclavine enhances natural killer (NK) cell activity, increases interleukin-2 and interferon-gamma production and prolongs the survival time of tumor-bearing mice. The aim of this study was 1) to test the effect of agroclavine on NK activity in vivo, and 2) to assess the potential toxicity of high doses of agroclavine on cardiac and liver functions using creatine kinase MB (CKMB) and alanine aminotransferase (ALT) as biochemical markers in normal and stressed animals. The effect of stress was studied because we examined promising anticancer properties of agroclavine and malignant diseases are supposed to be a potent stressful event for patients. In our experiments 3-month-old male rats of the Wistar-Kyoto strain were used. Agroclavine was injected intraperitoneally (0.5 mg/kg or 0.05 mg/kg) 30 min before stress (four hours' restraint and immersion in 23 degrees C water). The animals were killed 30 min after stress, blood was collected and the spleen was removed. Non-stressed animals treated with agroclavine were killed 5 h after the drug administration. The results confirmed our previous in vitro results and showed that also in vivo agroclavine increases NK cell activity under non-stress conditions. Agroclavine only slightly increased CKMB and had no influence on ALT in non-stressed animals. These promising results are limited by the fact that agroclavine (0.5 mg/kg) diminished NK cell activity and significantly increased ALT and CKMB under stress conditions.

Human BAC ends quality assessment and sequence analyses.

End sequences from bacterial artificial chromosomes (BACs) provide highly specific sequence markers in large-scale sequencing projects. To date, we have generated >300,000 end sequences from >186,000 human BAC clones with an average read length of >460 bp for a total of 141 Mb covering approximately 4.7% of the genome. Over 60% of the clones have BAC end sequences (BESs) from both ends representing more than fivefold coverage of the human genome by the paired-end clones. Our quality assessments and sequence analyses indicate that BESs from human BAC libraries developed at The California Institute of Technology (CalTech) and Roswell Park Cancer Institute have similar properties. The analyses have highlighted differences in insert size for different segments of the CalTech library. Problems with the fidelity of tracking of sequence data back to physical clones have been observed in some subsets of the overall BES dataset. The annotation results of BESs for the contents of available genomic sequences, sequence tagged sites, expressed sequence tags, protein encoding regions, and repeats indicate that this resource will be valuable in many areas of genome research.

Transcriptional regulation of the proton-translocating ATPase (atpIBEFHAGDC) operon of Escherichia coli: control by cell growth rate.

The F0F1 proton-translocating ATPase complex of Escherichia coli, encoded by the atpIBEFHAGDC operon, catalyzes the synthesis of ATP from ADP and Pi during aerobic and anaerobic growth when respiratory substrates are present. It can also catalyze the reverse reaction to hydrolyze ATP during nonrespiratory conditions (i.e., during fermentation of simple sugars) in order to maintain a electrochemical proton gradient across the cytoplasmic membrane. To examine how the atp genes are expressed under different conditions of cell culture, atpI-lacZ operon fusions were constructed and analyzed in single copy on the bacterial chromosome or on low-copy-number plasmids. Expression varied over a relatively narrow range (about threefold) regardless of the complexity of the cell growth medium, the availability of different electron acceptors or carbon compounds, or the pH of the culture medium. In contrast to prior proposals, atp operon expression was shown to occur from a single promoter located immediately before atpI rather than from within it. The results of continuous-culture experiments suggest that the cell growth rate rather than the type of carbon compound used for growth is the major variable in controlling atp gene expression. Together, these studies establish that synthesis of the F0F1 ATPase is not greatly varied by modulating atp operon transcription.