Isolation and identification of Streptomyces sp. Act4Zk, a good producer of Staurosporine and some derivatives.

In this study, strain Streptomyces sp. Act4Zk was isolated based on a method developed for the isolation of myxobacteria. Due to the low efficiency of the majority of conventional DNA extraction techniques, for molecular identification of the strain Streptomyces sp. Act4Zk, a new technique for DNA extraction of Actinobacteria was developed. In order to explore potential bioactivities of the strain, extracts of the fermented broth culture were prepared by an organic solvent (i.e. ethyl acetate) extraction method using. These ethyl acetate extracts were subjected to HPLC fractionation against standard micro-organisms, followed by LC/MS analysis. Based on morphological, physiological, biochemical and 16S rRNA gene sequence data, strain Streptomyces sp. Act4Zk is likely to be a new species of Streptomyces, close to Streptomyces genecies and Streptomyces roseolilacinus. Antimicrobial assay indicated high antifungal activity as well as antibacterial activity against Mycobacterium smegmatis and Gram-positive bacteria for the new strain. HPLC and LC/MS analyses of the extracts led to the identification of three different compounds and confirmed our hypothesis that the interesting species of the genus Streptomyces being a good producer of staurosporine and some derivatives.

Human adult cardiac autonomic innervation: Controversies in anatomical knowledge and relevance for cardiac neuromodulation.

BACKGROUND: Cardiac sympathetic blockade is a therapeutic approach for arrhythmias and heart failure and may be a beneficial effect of high thoracic epidural anesthesia. These treatments require detailed knowledge of the spatial location and distribution of cardiac autonomic nerves, however, there are controversies on this subject in humans. OBJECTIVE: To provide a systematic overview of current knowledge on human anatomy of the cardiac autonomic nervous system. RESULTS: In contrast to the often claimed assumption that human preganglionic sympathetic cardiac neurons originate mainly from thoracic spinal segments T1-T4 or T5, there is ample evidence indicating involvement of cervical spinal segment C8 and thoracic spinal segments below T5. Whether cervical ganglia besides the stellate ganglion play a role in transmission of cardiac sympathetic signals is unclear. Similarly, there is debate on the origin of cardiac nerves from different thoracic ganglia. Most human studies report thoracic cardiac nerves emerging from the first to fourth thoracic paravertebral ganglia; others report contributions from the fifth, sixth and even the seventh thoracic ganglia. There is no agreement on the precise composition of nerve plexuses at the cardiac level. After years of debate, it is generally accepted that the vagal nerve contributes to ventricular innervation. Vagal distribution appears higher in atria, whereas adrenergic fibers exceed the number of vagal fibers in the ventricles. CONCLUSION: Anatomy of the human cardiac autonomic nervous system is highly variable and likely extends beyond generally assumed boundaries. This information is relevant for thoracic epidural anesthesia and procedures targeting neuronal modulation of cardiac sympathetic innervation.

Acute kidney injury after open ventral hernia repair: an analysis of the 2005-2012 ACS-NSQIP datasets.

PURPOSE: Acute kidney injury (AKI) is a serious postoperative complication, negatively impacting mortality rates, extending length of stay, and raising hospital costs. The purpose of this study was to examine AKI following open ventral hernia repair (OVHR) using a large, heterogeneous database to determine the incidence and identify risk factors for this complication. METHODS: Using the 2005-2012 ACS-NSQIP database, patients undergoing open ventral hernia repair were identified by CPT codes. Patients with acute kidney injury within 30 days of surgery were compared to controls by multivariate logistic regression across preoperative and intraoperative characteristics. RESULTS: Of 48,629 open ventral hernia repair patients identified in the dataset, AKI developed in 1.4% (681 patients). Multivariate logistic regression determined a number of factors associated with AKI. These include WHO Class III obesity (OR = 2.57, p < 0.001), history of cardiovascular disease (OR = 1.81, p < 0.001), diabetes (OR = 1.29, p = 0.028), hypoalbuminemia (OR = 1.42, p = 0.004), and chronic kidney disease (for a baseline GFR of 60-89 mL/min/1.73 m2, OR = 1.62, p = 0.001; for 30-59 mL/min/1.73 m2, OR = 2.25, p < 0.001; for 15-29 mL/min/1.73 m2, OR = 4.96, p < 0.001). Intraoperative factors include prolonged operative time (for ≥1 SD above the mean, OR = 1.68, p = 0.002; for ≥2SD above the mean, OR = 2.76, p < 0.001) and intraoperative transfusion (OR = 2.44, p < 0.001). CONCLUSIONS: Patients with a history of obesity, chronic kidney disease, cardiovascular history, diabetes, and hypoalbuminemia are at increased risk for AKI when undergoing OVHR. Intraoperative variables such as prolonged operative times and blood transfusions may also suggest increased risk. Preoperative identification of patients with these characteristics and perioperative hemodynamic stabilization are important first steps to minimize this complication.

24 hour forecast of the surface UV for the antipsoriatic heliotherapy in Poland.

Analyses of the spectral and broad-band UV data collected at Belsk (20.79°E, 51.84°N), Poland, show that standard broad-band instrument, Solar Light (SL) 501A, could be used for measurements of both erythemal and antipsoriatic irradiance. A prognostic model is proposed for the next-day duration of outdoor exposure required to receive a dose, the so-called minimum antipsoriatic dose (MAD), equivalent to that received by standard antipsoriatic daily treatment in the phototherapy cabinet containing TL-01 fluorescent tubes. The model uses the 24 h forecast of the column amount of ozone (to predict next day clear sky UV irradiance), and low- and mid-level cloudiness (to estimate a reduction of the clear-sky UV irradiation due to clouds). The predicted duration of sunbathing required to receive a dose of 1 MAD matches the observed value, i.e. the correlation coefficients is 0.68. If the model predicts the antipsoriatic exposure over 1 MAD threshold the observed dose will be also above this threshold in 91% of cases. Thus, the model could be used for planning the next-day outdoor exposure to clear psoriasis. Hourly resolved maps, starting from 6 am up to 1 pm (GMT), showing the duration of antipsoriatic exposure over Poland are made public. The model provides a tool for a psoriatic patient to find the sunbathing starting time and its duration, which has the same healing potential as a single indoor phototherapy session.

Readmission following ventral hernia repair: a model derived from the ACS-NSQIP datasets.

BACKGROUND: Institutions are now incentivized to decrease rates of preventable readmissions. The purpose of this study was to examine readmissions following open ventral hernia repair (VHR), to ultimately create a model to preoperatively identify high-risk patients. STUDY DESIGN: Utilizing the 2011 and 2012 ACS-NSQIP datasets, patients undergoing open VHR were identified by CPT codes. Patients who were readmitted in 2011 within 30 days of the procedure were compared to those who were not with regard to preoperative and operative characteristics. A bootstrap analysis was performed to identify internally validated risk factors to be included in the final logistic regression, which was utilized to create a weighted model to predict the risk of readmission. This model was then validated with VHR patients in 2012. RESULTS: Overall, 10,745 patients were included for model generation. Of these, 850 (7.9%) patients were readmitted within 30 days. The final bootstrap analysis demonstrated that active smoking, ASA ≥ 3, a history of bleeding disorder or anemia, long operative time, inpatient status, and concurrent panniculectomy were all independently associated with readmission following ventral hernia repair. Significant variables were assigned a weighted score, ranging from 1 to 3. Each patient was then placed into one of four cohorts according to their summed score. The internally validated model [Hernia Readmission Risk (HERR) Score] demonstrated that risk increased in a linear fashion, with the highest risk cohort having a 21% risk of 30-day readmission. CONCLUSIONS: Perioperative predictors of readmission following VHR include smoking, ASA score, operative magnitude, concurrent panniculectomy, and preoperative anemia and bleeding disorders. The presented model based on these factors can aid in perioperative risk stratification for readmission.

Wound risk assessment in ventral hernia repair: generation and internal validation of a risk stratification system using the ACS-NSQIP.

BACKGROUND: There is a need for validated risk models to better stratify surgical site occurrences (SSO) following open ventral hernia repair (OVHR). The addition of more generalizable and validated risk models will serve to improve perioperative care in OVHR patients. METHODS: We reviewed the 2005-2011 ACS-NSQIP databases identifying encounters for OVHR. The dependent outcome measure of interest was SSO, defined as superficial surgical site infection, deep infection, organ space infection, or wound dehiscence. Multivariate logistic regression of independently associated factors was performed and internally validated using a bootstrap technique. A composite risk score, the Hernia Wound Risk Assessment Tool (HW-RAT) was created using weighted beta coefficients. The HW-RAT was compared to existing models from the literature. RESULTS: A total of 60,187 patients who met inclusion criteria were identified in the 2005-2011 ACS-NSQIP databases. The incidence of SSO in the study was 6.2% (N = 3,732). SSO risk factors were broken down based on rounded risk scores into the following groups: mild, intermediate, moderate, and severe risk. Severe risk factors related to operative time and degree of wound contamination. Moderate risk factors included class III obesity, component separation, dependent functional status, and inpatient hernia surgery. Patient stratification was performed based on total risk score into HW-RAT risk groups 1 through 5 which demonstrated significant discrimination between and across each group (P < 0.01, C-statistic = 0.71) with an incidence of SSO that ranged from 3.3 to 26.5%. CONCLUSION: We present an internally validated risk model of SSO in OVHR (HW-RAT), which complements and builds upon current risk models. LEVEL OF EVIDENCE: Prognostic/risk category, level II.

Perspectives of the antipsoriatic heliotherapy in Poland.

Statistical analysis of the daily course of exposures to TL-01 tube radiation for 93 psoriatic patients from the Medical University of Lódz during 20-day phototherapy shows that the dose of 1 J/cm(2) represents a unit of single exposure necessary for psoriasis healing. This value is converted to the antipsoriatic effective dose of 317.9 J/m(2) using the TL-01 lamp irradiance spectrum and the antipsoriatic action spectrum. It is proposed that the daily exposure of 317.9 J/m(2) serves as the standard antipsoriatic dose (SAPD) providing a link between the cabinet and the out-door exposures and it could be used for planning heliotherapy in Poland. A model is proposed to calculate ambient antipsoriatic doses for 3 h exposures around the local noon (9 am-12 am GMT) based on satellite measurements of ozone and cloud characteristics. The model constants are determined by a comparison with pertaining antipsoriatic doses measured by the Brewer spectrophotometer in central Poland. It is found that 3 h exposures to solar radiation in the period 15 May-15 September provides the mean (2005-2013) doses in the range 2.7-3.1 SAPD over Poland. Thus, heliotherapy could be treated as an alternative to the cabinet phototherapy for almost 4 months. It seems that the most effective site for antipsoriatic heliotherapy is the south/east part of Poland (the Bieszczady Mountains). The heliotherapy could be carried out in existing national health centers equipped with the standard easy-to-use biometers for on-line monitoring of UV level and controlling duration of sunbathing to avoid erythema risks. It is even possible to control the antipsoriatic heliotherapy by a patient himself, using low-cost hand-held instruments measuring UV index.

Upper thoracic epidural anaesthesia: effects of age on neural blockade and cardiovascular parameters.

BACKGROUND: Segmental dose reduction with increasing age after thoracic epidural anaesthesia (TEA) has been documented. We hypothesised that after a fixed loading dose of ropivacaine at the T3-T4 level, increasing age would result in more extended analgesic spread. In addition, other aspects of neural blockade and haemodynamic changes were studied. METHODS: Thirty-five lung surgery patients were included in three age groups. Thirty-one patients received an epidural catheter at the T3-T4 interspace followed by an injection of 8-ml ropivacaine 0.75%. Analgesia was assessed with pinprick and temperature discrimination. Motor block was tested using the Bromage and epidural scoring scale for arm movements score. An arterial line was inserted for invasive measurement of blood pressure, cardiac index (CI) and stroke volume (SV). RESULTS: There was no influence of age on quality of TEA except for the caudal border of analgesia being somewhat lower in the middle and older age group compared with the young age group. Heart rate (6.0 ± 5.9, P < 0.001), mean arterial pressure (16.1 ± 15.6, P < 0.001), CI (0.55 ± 0.49, P < 0.001) and SV (9.6 ± 14.6, P = 0.001) decreased after TEA for the total group. Maximal reduction in heart rate after TEA was more extensive in the young age group compared with the other age groups. There was no effect of age on other cardiovascular parameters. CONCLUSION: We were unable to demonstrate an effect of age on the maximal number of spinal segments blocked after TEA; however, the caudad spread of analgesia increased with advancing age. In addition, reduction of heart rate was greater in the youngest group.

Molecular dissection of the chromosome band 7q21 amplicon in gastroesophageal junction adenocarcinomas identifies cyclin-dependent kinase 6 at both genomic and protein expression levels.

Amplification of chromosome band 7q21 has been frequently detected in various types of cancer including gastroesophageal junction (GEJ) adenocarcinomas. At present, no gene has been disclosed that can explain this frequent amplification of 7q21 in GEJ carcinomas. Therefore, a detailed genomic analysis of the 7q21 region was performed on a selected series of GEJ adenocarcinomas, i.e., 14 primary adenocarcinomas and 10 cell lines, by array comparative genomic hybridization (aCGH) with a 7q11.22-q31.2 contig array. A distinct peak of amplification was identified at 92.1 Mb in 7q21.2, precisely comprising cyclin-dependent kinase 6 (CDK6), a gene involved in cell cycle regulation. A smaller peak was seen at 116.2 Mb in 7q31.2, the locus of the MET proto-oncogene. No distinct peak was detected for the hepatocyte growth factor (HGF) at 81.3 Mb in 7q21.11. An immunoprofile of HGF, CDK6 and MET revealed a strong correlation between aCGH and immunohistochemical protein expression for CDK6 (P = 0.002). Furthermore, immunohistochemistry did not show expression of CDK6 in Barrett's dysplasia and carcinoma in situ, correlating expression of CDK6 with a malignant phenotype. We conclude that high-resolution genomic analysis and immunoprofiling identify CDK6 as the main candidate target for the recurrent amplification of 7q21 in GEJ adenocarcinomas.

Cyclipostins: novel hormone-sensitive lipase inhibitors from Streptomyces sp. DSM 13381. I. Taxonomic studies of the producer microorganism and fermentation results.

The cyclipostins are a group of hormone-sensitive lipase inhibitors produced by a Streptomyces species. Having verticillate spore chains this strain exhibits significant differences to the known species of the former genus Streptoverticillium. Taxonomic studies and fermentation results are presented.

Pluraflavins, potent antitumor antibiotics from Saccharothrix sp. DSM 12931.

The new pluramycin-type antibiotics pluraflavin A, C43H54N2O14, pluraflavin B, C43H56N2O15, and pluraflavin E, C36H41NO14 were isolated from cultures of the Saccharothrix species DSM 12931. The structures of the novel compounds were elucidated with the aid of 2D NMR and mass spectrometric investigations. The characteristic structural element of pluraflavins A and B is an additional 4-epi-vancosamine unit at position 13 of the anthraquinone-gamma-pyrone ring system. Pluraflavin E has a carboxyl group in this position. Pluraflavin A has a reactive dimethyl epoxide side chain at position 2 of the anthraquinone-gamma-pyrone aglycon, which may explain the high activity of the antibiotic. The outstanding biological characteristic of pluraflavin A is its powerful, organ-dependent cytostatic action: the IC50 in the colon carcinoma proliferation assay is in the subnanomolar range.

Nonomuraea dietzii sp. nov..

The taxon 'Sebekia benihana', an actinomycete of industrial significance, has been subjected to a polyphasic taxonomic study. 16S rDNA sequence analyses of strain DSM 44320 (= NRRL 11111) revealed 100% similarity to the 16S rDNA sequence of Nonomuraea roseola DSM 43767T and high values (>98.5%) to some other species of this genus. DNA-DNA similarity values of less than 31% observed between strain DSM 44320 and N. roseola DSM 43767T, Nonomuraea recticatena DSM 43937T and Nonomuraea africana DSM 43748T indicate that this strain represents an individual genospecies. Differences between strain DSM 44320T and the 16 validly described Nonomuraea species occur in the primary structure of the 16S rDNA, the physiological test profile, the morphology of mycelium and the spore surface and the colour of substrate mycelium. The only strain of 'S. benihana', DSM 44320T (= NRRL 11111T), is proposed as the type strain of a new species of the genus Nonomuraea, Nonomuraea dietzii sp. nov..

Cephaibols, new peptaibol antibiotics with anthelmintic properties from Acremonium tubakii DSM 12774.

Two groups of new peptaibol-type antibiotics termed cephaibols have been isolated from the fungus Acremonium tubakii, DSM 12774. These 16- or 17-unit straight-chain peptides, whose structures were characterized by amino acid analyses, 2-D NMR experiments, and by mass spectrometric sequencing, have a high content of the unusual amino acids aminoisobutyric acid and isovaline. The principal constituent of the novel peptaibol mixture is cephaibol A, which is formed in abundance in cultures of the wild strain. The striking biological property of cephaibol A is its pronounced anthelmintic action and activity against ectoparasites.

Memnopeptide A, a novel terpene peptide from Memnoniella with an activating effect on SERCA2.

The terpene peptide memnopeptide A (1), C76H108N16O18S, MW 1564, was isolated from a culture of the fungus Memnoniella echinata FH 2272 on casein peptone. The structure of the novel compound was elucidated with the aid of 2D NMR experiments and from amino acid analysis and mass spectrometric sequencing of the peptide. The compound consists of a known phenylspirodrimane subunit linked to the decapeptide Met-His-Gln-Pro-His-Gln-Pro-Leu-Pro-Pro. This proline-rich peptide is a subsequence of beta-casein. From the observed absence in the literature of any other highly significant sequence homologues, memnopeptide A can be assumed to arise from metabolic products of the fungus with direct incorporation of constituents of the nutrient medium. The formation of memnopeptide A suggests this may be a mechanism for storage of amines by the fungus. Memnopeptide A has weak antibacterial activity against gram-positive bacteria and effects half-maximal activation of sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA2) at a concentration of 12.5 microM.

Friulimicins: novel lipopeptide antibiotics with peptidoglycan synthesis inhibiting activity from Actinoplanes friuliensis sp. nov. I. Taxonomic studies of the producing microorganism and fermentation.

A strain that produces new lipopeptide antibiotics is a new species of the genus Actinoplanes for which we propose the name Actinoplanes friuliensis (type strain: HAG 010964). The strain is an actinoplanete actinomycete having cell wall II composition and forming sporangia. Comparisons with Actinoplanes spp. which have similarities with our isolate, including fatty acid analysis, showed that the isolate belongs to a new species. Taxonomic studies and fermentation are presented.

Molecular cytogenetic evaluation of virus-associated and non-viral hepatocellular carcinoma: analysis of 26 carcinomas and 12 concurrent dysplasias.

The worldwide incidence of hepatocellular carcinoma (HCC) is approximately one million cases a year. This makes HCC one of the most frequent human malignancies, especially in Asia and Africa, although the incidence is increasing also in the western world. HCC is a complication of chronic liver disease, with cirrhosis as the most important risk factor. Viral co-pathogenesis makes cirrhosis due to hepatitis B (HBV) and hepatitis C virus (HCV) infection a very important factor in the development of HCC. As curative therapy is often ruled out due to the late detection of HCC, it would be attractive to find parameters which predict malignant transformation in HBV- and HCV-infected livers. This study has used comparative genomic hybridization (CGH) to analyse 26 HCCs (11 non-viral, nine HBV, six HCV) and 12 concurrent dysplasias (five non-viral, five HBV, two HCV). Frequent gain (> or =25% of all tumours) was detected, in decreasing order of frequency, on 8q (69%), 1q (46%), 17q (46%), 12q (42%), 20q (31%), 5p (27%), 6q (27%), and Xq (27%). Frequent loss (> or =25% of all tumours) was found, in decreasing order of frequency, on 8p (58%), 16q (54%), 4q (42%), 13q (39%), 1p (35%), 4p (35%), 16p (35%), 18q (35%), 14q (31%), 17p (31%), 9p (27%), and 9q (27%). Minimal overlapping regions could be determined at multiple locations (candidate genes in parentheses). Minimal regions of overlap for deletions were assigned to 4p14-15 (PCDH7), 8p21-22 (FEZ1), 9p12-13, 13q14-31 (RB1), 14q31 (TSHR), 16p12-13.1 (GSPT1), 16q21-23 (CDH1), 17p12-13 (TP53), and 18q21-22 (DPC4, DCC). Minimal overlapping amplified sites could be seen at 8q24 (MYC), 12q15-21 (MDM2), 17q22-25 (SSTR2, GH1), and 20q12-13.2 (MYBL2, PTPN1). A single high level amplification was seen on 5q21 in an HBV-related tumour. Aberrations appeared more frequent in HBV-related HCCs than in HCV-associated tumours (p=0.008). This was most prominent with respect to losses (p=0.004), specifically loss on 4p (p=0.007), 16q (p=0.04), 17p (p=0.04), and 18q (p=0.03). In addition, loss on 17p was significantly lower in non-viral cancers than in HBV-related HCC (p<0.001). Furthermore, loss on 13q was more prevalent in HCCs in non-cirrhotic livers (p=0.02), thus suggesting a different, potentially more aggressive, pathway in neoplastic progression. A tendency (p=0.07) was observed for loss on 9q in high-stage tumours; no specific changes were found in relation to tumour grade. A subset of the HCC-associated genetic changes was disclosed in the preneoplastic stage, i.e. liver cell dysplasia. This group of dysplasias showed frequent gain on 17q (25%) and frequent loss on 16q (33%), 4q (25%), and 17p (25%). The majority of the dysplasias with alterations revealed genetic changes that were also present in the primary tumour. In conclusion, firstly, this study has provided a detailed map of genomic changes occurring in HCC of viral and non-viral origin, and has suggested candidate genes. Loss on 17p, including the TP53 region, appeared significantly more prevalent in HBV-associated liver cancers, whereas loss on 13q, with possible involvement of RB1, was distinguished as a possible genetic biomarker. Secondly, CGH analysis of liver cell dysplasia, both viral and non-viral, has revealed HCC-specific early genetic changes, thereby confirming its preneoplastic nature. Finally, genes residing in these early altered regions, such as CDH1 or TP53, might be associated with hepatocellular carcinogenesis.

Ala(0)-actagardine, a new lantibiotic from cultures of Actinoplanes liguriae ATCC 31048.

The actagardine-producing strain Actinoplanes liguriae ATCC 31048, forms an additional lantibiotic when it is cultured on mannitol and soya meal. The new compound, Ala(0)-actagardine (1), has been isolated by solid-phase extraction followed by a two-step chromatographic separation. The molecular formula of 1 is C84H129N21O25S4. Its chemical structure was determined by 2D-NMR analysis and was further confirmed by an amino acid analysis, Edman degradation, and partial synthesis from actagardine. 1 exhibits a slightly higher biological activity than the parent compound actagardine. The synthetic analogs Lys(0)-actagardine (2) and Ile(0)-actagardine (3) demonstrate also antibacterial activities and emphasize the importance of the N-terminus for further derivatization.