GRP78-targeting subtilase cytotoxin sensitizes cancer cells to photodynamic therapy.
Glucose-regulated protein 78 (GRP78) is an endoplasmic reticulum (ER)-resident chaperone and a major regulator of the unfolded protein response (UPR). Accumulating evidence indicate that GRP78 is overexpressed in many cancer cell lines, and contributes to the invasion and metastasis in many human tumors. Besides, GRP78 upregulation is detected in response to different ER stress-inducing anticancer therapies, including photodynamic therapy (PDT). This study demonstrates that GRP78 mRNA and protein levels are elevated in response to PDT in various cancer cell lines. Stable overexpression of GRP78 confers resistance to PDT substantiating its cytoprotective role. Moreover, GRP78-targeting subtilase cytotoxin catalytic subunit fused with epidermal growth factor (EGF-SubA) sensitizes various cancer cells to Photofrin-mediated PDT. The combination treatment is cytotoxic to apoptosis-competent SW-900 lung cancer cells, as well as to Bax-deficient and apoptosis-resistant DU-145 prostate cancer cells. In these cells, PDT and EGF-SubA cytotoxin induce protein kinase R-like ER kinase and inositol-requiring enzyme 1 branches of UPR and also increase the level of C/EBP (CCAAT/enhancer-binding protein) homologous protein, an ER stress-associated apoptosis-promoting transcription factor. Although some apoptotic events such as disruption of mitochondrial membrane and caspase activation are detected after PDT, there is no phosphatidylserine plasma membrane externalization or DNA fragmentation, suggesting that in DU-145 cells the late apoptotic events are missing. Moreover, in SW-900 cells, EGF-SubA cytotoxin potentiates PDT-mediated cell death but attenuates PDT-induced apoptosis. In addition, the cell death cannot be reversed by caspase inhibitor z-VAD, confirming that apoptosis is not a major cell death mode triggered by the combination therapy. Moreover, no typical features of necrotic or autophagic cell death are recognized. Instead, an extensive cellular vacuolation of ER origin is observed. Altogether, these findings indicate that PDT and GRP78-targeting cytotoxin treatment can efficiently kill cancer cells independent on their apoptotic competence and triggers an atypical, non-apoptotic cell death.


Role of nitric oxide in control of growth hormone release in the rat.
Previous experiments in this and other laboratories have revealed that nitric oxids (NO) plays a role in controlling the release of corticotropin-releasing hormone (CRH) and luteinizing-hormone-releasing hormone (LHRH). Therefore, we have investigated its role in control of growth hormone (GH) release in conscious rats by microinjecting NG-monomethyl-L-arginine (NMMA), an inhibitor of NO synthase (NOS), into the third ventricle (3V) of conscious, freely moving castrate male rats. An initial blood sample (0.3 ml) was drawn from an indwelling intra-atrial catheter just prior to injection of NMMA [1 mg in 5 microliters of 0.9% NaCl (saline)] into the 3V. To maintain the inhibitory action on NOS, a second injection of NMMA was administered into the 3V 60 min after the first. Additional blood samples (0.3 ml) were removed at 10 min intervals for 120 min. Other animals received injections of the diluent at the same times and volumes as NMMA. Interleukin (IL)-1 alpha (0.06 pmol in 2 microliters saline) was injected into the 3V immediately after the first injection of NMMA, whereas other animals received the NMMA diluent followed by IL-1 alpha. The effects of IL-1 alpha were almost identical to those of NMMA in that there was a dramatic lowering of plasma GH achieved primarily by a reduction in height of the GH pulses without a significant reduction in their number.(ABSTRACT TRUNCATED AT 250 WORDS)


Cervicogenic headache: anatomic basis and pathophysiologic mechanisms.
Cervicogenic headache is pain perceived in the head but referred from a primary source in the cervical spine. The physiologic basis for this pain is convergence between trigeminal afferents and afferents from the upper three cervical spinal nerves. The possible sources of cervicogenic headache lie in the structures innervated by the C1 to C3 spinal nerves, and include the upper cervical synovial joints, the upper cervical muscles, the C2-3 disc, the vertebral and internal carotid arteries, and the dura mater of the upper spinal cord and posterior cranial fossa. Experiments in normal volunteers have established that the cervical muscles and joints can be sources of headache.


Low back pain and illness behavior (inappropriate, maladaptive, or abnormal).
Patients with low back pain without a sufficient physical cause attract labels such as "hysteria" and "hypochondriasis," often on the basis of exclusion. The concept of "abnormal illness behavior" was introduced to clarify the classification and diagnosis of these conditions, in which the behavior of the doctor plays a particularly important part. Principles of management are discussed.


The Dry-aging and Heating Effects on Protein Characteristics of Beef Longissiumus Dorsi.
The aim of this study was to investigate the effects of dry-aging (DA) and the cooking process on the myofibril protein functionalities and in vitro digestibility of proteins in beef loin. Six sirloins from beef were dry-aged for 28 d, and the control group (n=6) was analyzed 2 d postmortem for this study. Dimensional changes (reduction of thickness and surface shrinkage) after cooking were significantly greater in the control group than the DA group, whereas the shear force of the DA group was significantly lower than that of the control. Effect of cooking on aggregation, hydrophobicity, and in vitro digestibility were significantly higher in the DA group than in the control. After cooking, the protein in DA sirloins was more oxidized than in the control samples. According to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis result, the low molecular weight bands (below 17 kDa) increased in the DA group, finding that the protein characteristics of dry-aged beef was affected by cooking.


Risk factors of digital dermatitis in feedlot cattle.
Digital dermatitis (DD) has been reported in North American feedlots, although risk factors are not well characterized. Our objectives were to analyze: (1) foot and leg conformation and (2) pen hygiene, as potential variables that predispose feedlot cattle to DD. Production parameters in DD-affected cattle were compared with healthy cattle and with those diagnosed with more commonly known infectious lesion foot rot (FR). In total, 2,854 feedlot cattle in 11 pens in 2 feedlots were assessed (bi-weekly pen walks) throughout the feeding cycle. Pen condition was categorized as: "dry," "mud present but has good bedding," "more mud than bedding," and "excessive mud." Gait scoring was competed and cattle with abnormal gait or evident foot lesions (i.e., DD or FR) were restrained in a cattle chute for a close foot inspection (n=280), including scoring of foot angle and claw set and hind and side views of rear feet and legs. Cumulative incidence of DD (present or absent) and FR was 2.5% (71/2,854) and 11.6% (331/2,854), respectively. Foot and leg conformation was not significantly different between left and right sides or between cattle with (n=71) and without DD (n=209). Lameness was diagnosed in only 22% of cattle with DD. Cattle with DD gained 0.27 kg/d less compared with healthy cattle (mean ± SD: 1.29 ± 0.29 vs. 1.56 ± 0.27, P<0.05) and 0.4 kg/d less compared with FR (1.29 ± 0.29 vs. 1.69 ± 0.25). Presence of DD was not significantly different between pens with "dry" and "mud present but has good bedding," but for pens with "more mud than bedding" or "excessive mud," the risk of cattle having DD cases increased significantly [odds ratio (OR)=8.55, confidence interval (CI): 4.0-18.4 and OR=14.1, CI: 5.9-33.8, respectively]. In conclusion, it is important to keep good pen conditions to reduce the risk of DD, which can be managed through proper stocking density and strategic bedding, irrespective of foot and leg conformation.


Bacillus xiamenensis sp. nov., isolated from intestinal tract contents of a flathead mullet (Mugil cephalus).
A taxonomic study was carried out on strain HYC-10(T), which was isolated from the intestinal tract contents of a flathead mullet, Mugil cephalus, captured from the sea off Xiamen Island, China. The bacterium was observed to be Gram positive, oxidase and catalase positive, rod shaped, and motile by subpolar flagella. The bacterium was found to grow at salinities of 0-12 % and at temperatures of 8-45 °C. The isolate was found to hydrolyze aesculin and gelatin, but was unable to reduce nitrate to nitrite. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain HYC-10(T) belongs to the genus Bacillus, with highest sequence similarity (99.3 %) to Bacillus aerophilus 28K(T), Bacillus stratosphericus 41KF2a(T) and Bacillus altitudinis DSM 21631(T), followed by Bacillus safensis DSM 19292(T) (99.5 %) and Bacillus pumilus DSM 27(T) (99.5 %), while the sequence similarities to others were all below 97.6 %. The genomic ANIm values between strain HYC-10(T) and three type strains (B. altitudinis DSM 21631(T), B. safensis DSM 19292(T) and B. pumilus DSM 27(T)) were determined to range from 89.11 to 91.53 %. The DNA-DNA hybridization estimate values between strain HYC-10(T) and the three type strains were from 36.60 to 44.00 %. The principal fatty acids identified were iso-C15:0 (39.1 %), anteiso-C15:0 (22.7 %), iso-C17:0 (13.1 %), C16:0 (6.1 %), anteiso-C17:0 (5.8 %) and iso-C16:0 (5.1 %). The G+C content of the chromosomal DNA was determined from the draft genome sequence to be 41.3 mol%. The respiratory quinone was determined to be MK-7 (100 %). Phosphatidylglycerol, diphosphatidylglycerol, aminoglycolipid, two glycolipids and two unknown phospholipids were found to be present. The combined genotypic and phenotypic data show that strain HYC-10(T) represents a novel species of the genus Bacillus, for which the name Bacillus xiamenensis sp. nov. is proposed, with the type strain HYC-10(T) (=CGMCC NO.1.12326(T) = LMG 27143(T) = MCCC 1A00008(T)).


Is Robot-Assisted Surgery Really Scarless Surgery? Immediate Reconstruction with a Jejunal Free Flap for Esophageal Rupture after Robot-Assisted Thyroidectomy.
Esophageal perforation is a rare but potentially fatal complication of robot-assisted thyroidectomy (RAT). Herein, we report the long-term outcome of an esophageal reconstruction with a jejunal free flap for esophageal rupture after RAT. A 33-year-old woman developed subcutaneous emphysema and hoarseness on postoperative day1 following RAT. Esophageal rupture was diagnosed by computed tomography and endoscopy, and immediate surgical exploration confirmed esophageal rupture, as well as recurrent laryngeal nerve injury. We performed a jejunal free flap repair of the 8-cm defect in the esophagus. End-to-side microvascular anastomoses were created between the right external carotid artery and the jejunal branches of the superior mesenteric artery, and end-to-end anastomosis was performed between the external jugular vein and the jejunal vein. The right recurrent laryngeal nerve injury was repaired with a 4-cm nerve graft from the right ansa cervicalis. Esophagography at 1 year after surgery confirmed that there were no leaks or structures, endoscopy at 1 year confirmed the resolution of vocal cord paralysis, and there were no residual problems with swallowing or speech at a 5-year follow-up examination. RAT requires experienced surgeons with a thorough knowledge of anatomy, as well as adequate resources to quickly and competently address potentially severe complications such as esophageal rupture.


Event-related brain potentials in elderly dippers and nondippers with recently diagnosed hypertension.
Several studies have shown a relationship between blood pressure (BP) and cognitive function. Yet very few studies have addressed circadian BP patterns in this context, perhaps due to poor availability of suitable methods to detect slight changes in the cognitive state. Today, brain event-related potentials (ERPs) allow us to detect subclinical changes in cognitive function. We enrolled 30 elderly patients with recently diagnosed hypertension (<2 years) that had never been treated: 18 dippers and 12 nondippers. Patients underwent 24-h ambulatory blood pressure monitoring (ABPM). Careful assessment of their cognitive state was carried out using the mini mental state examination (MMSE), and the recording of P300 and N2 ERPs. No significant differences between the two groups were found. MMSE scores in dippers and nondippers were similar (29.5+/-0.71 vs. 29.3+/-1.07, respectively; p =0.611), as were P300 latency values (377.78+/-33.28 vs. 364.67+/-35.12 in the central (Cz) position, p =0.310; 379.22+/-32.94 vs. 365.25+/-35.07 in the occipital (Pz) position, p =0.277) and N2 wave latency values (253.83+/-24.9 vs. 249.17+/-24.47 in the Cz position, p =0.617; 251.56+/-25.86 vs. 246.58+/-25.46 in the Pz position, p =0.608). These data show no association between the nondipping pattern and lower cognitive function in elderly subjects with recent hypertension.


[Use of hospital discharge records to estimate the incidence of malignant mesotheliomas].
cancer registries usually adopt strategies for active case finding. Interest in using administrative sources of data is rising to assess the usefullness of Hospital discharge records (HDR) to supplement the traditional methods of case finding of the malignant mesothelioma (MM) Registry of the Piedmont Region. HDRs have been used since 1996. We assessed the number of cases identified only through HDRs and their influence on MM incidence. cases identified through HDRs were about 10% of those with histologic confirmation of the diagnosis, 34% of those with cytologic confirmation, and 72% of those without morphologic examination. Cases diagnosed in hospitals located outside the region would have been easily (50%) missed. The age-standardised (standard: Italian pop. at the 1981 census) incidence rate of pleural MM increases from 2.2 to 2.7 per 100,000 per year among men, and from 1.1 to 1.2 among women, when including all cases identified from HDRs, irrespective of their diagnostic confirmation. Peritoneal MM incidence estimates are unaffected. Overall without access to the hospital discharge files, 179 cases out of 954 would not have been registered between 1996 and 2001. In the same calendar period 59 cases identified by means of active search by the Registry have not been found in the hospital discharge files. HDRs are useful in addition, but not in substitution, to active search of MM cases.


Treatment of metastatic prostate adenocarcinoma to the calcaneus.
Metastatic skeletal adenocarcinoma is an all too common and unfortunate complication of advanced oncologic states. Mortality rates are usually elevated when bony metastasis are evident, as this signifies advanced disease. The foot and ankle are uncommon sites for metastatic deposits, but may occur. As such, the foot and ankle surgeon must be aware of the potential for such disease, and be able to proceed with an imaging and medical work-up of the patient with foot and ankle skeletal metastasis. The goal of treatment is pain relief and the preservation of functional ambulation, which may greatly enhance the quality of remaining life for patients. A team approach is mandatory to manage the patients with metastatic disease. We present a case of an elderly male with a known history of prostate cancer, who presented with unrelenting heel pain, which upon diagnostic work-up, proved to be progressive calcaneal as well as axial metastasis after a brief period of clinical remission. Operative management coupled with palliative radiation and bisphosphonate therapy provided symptomatic pain relief and maintenance of functional ambulation.


Changes in Cognitive Performance With Age in Adults With Multiple Sclerosis.
Cognitive impairment is one of the most common consequences of multiple sclerosis (MS), yet there is a shortage of data regarding how cognition changes during the life span of individuals with MS. This information is of increasing importance given the growing proportion of older adults with MS. To study possible changes in cognitive function in correlation with increasing age in individuals with MS. Participants (N=129) were recruited and a priori allocated into one of three age groups (young, middle-aged, and older). All participants completed the Brief International Cognitive Assessment for Multiple Sclerosis (BICAMS) during a single laboratory testing session. The BICAMS measures cognitive processing speed as well as verbal and visuospatial learning and memory. A multivariate analysis of variance indicated that cognitive function significantly differed by age group, and these differences were not explained by amount of physical activity, years of education, years since diagnosis, or race. Older adults displayed significantly worse cognitive processing speed than young and middle-aged adults. The older and middle-aged adults also demonstrated significantly worse visuospatial learning and memory than the younger adults. Effect sizes indicated that cognitive processing speed and verbal learning and memory were more affected in late adulthood than early adulthood, whereas visuospatial learning and memory was affected similarly in early and late adulthood. Older adults with MS demonstrated significant impairments in cognitive function compared to young and middle-aged adults with MS. Future studies should determine the predictors of cognitive decline in this age cohort.


In-vivo bacterial adherence to intracardiac prosthetic materials: a new experimental model.
Bacterial colonization of inert artificial materials is a critical variable in the appearance of foreign body-centered infections. Hence, a new experimental endocarditis model was evaluated that enables quantitative in-vivo testing of bacterial adherence to prosthetic materials. In 53 rabbits, different patches of either Dacron (DAC; n = 16), glutaraldehyde-fixed pericardium (GAP; n = 18) or cryopreserved allograft (AG; n = 19) were threaded onto a Prolene suture and led through a stitch incision at the left atrium to the ventricle, perforating the mitral valve and leaving the heart at the outside of the left ventricle. The patch became caught in the valve tissue, creating mitral insufficiency. At 6 h after surgery, 6 x 10(6) colony-forming units (CFU) of Staphylococcus aureus were administered intravenously. Postmortem examination and further analysis were carried out 48 h after bacterial inoculation. Ten animals (two DAC, four GAP, four AG) died and were excluded. Subsequently, sterile patches were found in 6/14 animals of the DAC group, in 9/14 of the GAP group, and in 9/15 of the AG group. The mean numbers of cultured organisms in the remaining patches were 2.3 x 10(6) + 7.6 x 10(6)/mg (DAC), 1.8 x 10(4)+/-6.7 x 10(4)/mg (GAP) and 4.9 x 10(5)+/-1.0 x 10(6) CFU/mg (AG). Differences in the number of CFU were not significant for all groups. The above-described model allows reliable quantitative in-vivo testing of bacterial adherence to different prosthetic materials within a short time period. The results suggest that, with regard to avoiding the development of bacterial endocarditis, biomaterials have no advantage over Dacron.


Primary structure of protein moiety of Penicillium notatum phospholipase B deduced from the cDNA.
Phospholipase B has not yet been well defined. The most important points about this enzyme are its relationships with lysophospholipase and phospholipase A1. As reported [Saito, K., Sugatani, J. & Okumura, T. (1991) Methods Enzymol. 197, 446-456], Penicillium notatum phospholipase B is a glycoprotein with a molecular mass of 95 kDa and intrinsic lysophospholipase and phospholipase B activities; however, by endogenous proteolytic modification, its phospholipase B activity is lost almost completely, whereas its lysophospholipase activity remains unchanged. A cDNA library of P. notatum was screened by hybridization with two synthetic oligodeoxyribonucleotide probes, which corresponds to two different pentapeptides of the enzyme. A hybridization-positive clone, pPLB18, was isolated and its nucleotide sequence was determined. The deduced amino acid sequence was quite different from that found previously. Therefore, we rescreened the cDNA library with a Sau3AI fragment derived from pPLB18 and isolated a new clone, pPLB15. Comparison of the nucleotide sequences of pPLB15 and pPLB18 revealed that pPLB18 contained an insertion sequence of 53 bp. Consequently, the reading frame was open downstream for 603 amino acid residues. From the assigned sequence, it was deduced that the limited proteolysis occurred between Leu175 and Asp176; eight cysteine residues and 16 potential N-glycosylation sites were also found. No amino acid sequence similarity was found with other proteins, including other phospholipases.


Elevated serum leptin may be associated with disease activity and secondary osteoporosis in Chinese patients with rheumatoid arthritis.
Rheumatoid arthritis (RA) is a systemic chronic autoimmune disease in adults that is associated with significant joint issues and systemic inflammation. One of the signs of bone damage in RA is osteoporosis (OP). Leptin is an inflammatory protein that has been reported to be related to RA. The potential relationships among leptin, disease activity, and OP in Chinese patients with RA are not well known. In total, 245 patients with RA and 120 healthy controls were included in this study. Detailed data on the clinical characteristics and laboratory features were collected. Information about physical activity and functional status was recorded using specific questionnaires. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DXA). The MECALL castor-50-hf model X-ray scanner was used for the two-hand (including wrist) photographs. Serum leptin levels differed significantly between the RA group and healthy control subjects (1.27/3.29 vs. 0.17/0.24, Z=13.29, P<0.001). The positive rate of leptin protein in RA patients was 86.35%, which was higher than that in controls (19.55%) (χ2=28.51, P<0.001). Pearson's correlation test showed that morning stiffness, disease duration, joint swelling, joint tenderness, swollen joint count (SJC), tender joint count (TJC), health assessment questionnaire (HAQ) score, and Sharp-van der Heijde method (Sharp) score were positively correlated with the level of serum leptin (r=0.212, r=0.312, r=0.322, r=0.501, r=0.291, r=0.334, P<0.05). There was a clear increasing trend in the level of serum leptin according to the different disease activity scores and in the 28 joint activity (DAS28) groups (F=13.936, P<0.001). Elevated leptin was a risk factor for increased disease activity and OP according to logistic regression analysis. The median leptin level differed significantly between the normal bone mass group, osteopenia group, and OP group (P<0.001). An increased serum leptin level was a risk factor for RA-induced osteoporosis according to logistic regression analysis (P<0.001). These results suggest that the level of serum leptin is associated with disease activity and secondary OP among Chinese patients with RA. Key Points • Serum leptin levels in RA patients are higher than those in normal control group. • Leptin was associated with disease activity. • Leptin was associated with the occurrence of systemic osteoporosis and affects bone erosion in RA patients.


Analysis of nevirapine (NVP) resistance in Ugandan infants who were HIV infected despite receiving single-Dose (SD) NVP versus SD NVP plus daily NVP up to 6 weeks of age to prevent HIV vertical transmission.
Single-dose nevirapine (SD NVP) at birth plus NVP prophylaxis for the infant up to 6 weeks of age is superior to SD NVP alone for prevention of vertical transmission of human immunodeficiency virus (HIV) through breastfeeding. We analyzed NVP resistance in HIV-infected Ugandan infants who received either SD NVP or extended NVP prophylaxis. We tested plasma HIV by using a genotyping assay (ViroSeq; Celera Diagnostics), a phenotypic resistance assay (PhenoSense; Monogram Biosciences), and sensitive point mutation assay (LigAmp, for K103N, Y181C, and G190A). When infants were 6 weeks old, ViroSeq detected NVP resistance in a higher proportion of infants in the extended NVP arm than in the SD NVP arm (21 of 25 [84%] vs. 12 of 24 [50%]; P = .01). Similar results were obtained with LigAmp and PhenoSense. In both study arms, infants who were HIV infected at birth frequently had NVP resistance detected. In contrast, infants in the extended NVP arm who were HIV infected after birth were more likely to have resistance detected at 6 weeks, compared with infants in the SD NVP arm. The use of extended NVP prophylaxis was also associated with detection of NVP resistance by ViroSeq at 6 months (7 of 7 [100%] infants in the extended NVP arm had resistance detected, compared with 1 of 6 [16.7%] infants in the SD NVP arm; P = .005). The use of extended NVP prophylaxis was associated with increased selection for and persistence of NVP resistance in HIV-infected Ugandan infants.


Metal-free heterogeneous catalysis for sustainable chemistry.
The current established catalytic processes used in chemical industries use metals, in many cases precious metals, or metal oxides as catalysts. These are often energy-consuming and not highly selective, wasting resources and producing greenhouse gases. Metal-free heterogeneous catalysis using carbon or carbon nitride is an interesting alternative to some current industrialized chemical processes. Carbon and carbon nitride combine environmental acceptability with inexhaustible resources and allow a favorable management of energy with good thermal conductivity. Owing to lower reaction temperatures and increased selectivity, these catalysts could be candidates for green chemistry with low emission and an efficient use of the chemical feedstock. This Review highlights some recent promising activities and developments in heterogeneous catalysis using only carbon and carbon nitride as catalysts. The state-of-the-art and future challenges of metal-free heterogeneous catalysis are also discussed.


Prevalence of Escherichia coli O157 in cattle and swine in central Mexico.
Escherichia coli O157:H7 is a foodborne pathogenic bacterium that can reside undetected in the gastrointestinal tract of cattle because colonization by this bacterium is asymptomatic. Recent research has indicated that swine can carry and transmit this pathogen as well. The development of more advanced and sensitive detection techniques has improved the limit of detection and increased sensitivity for this important pathogen. This study was undertaken to determine the prevalence of E. coli O157 in cattle and swine in Mexico with the more sensitive detection technique of immunomagnetic bead separation. Samples (n = 60 per farm) were taken from four cattle and four swine farms (n = 240 cattle samples, n = 240 swine samples) located throughout central Mexico in October 2001. The prevalence of E. coli O157 was found to be only 1.25% on cattle farms and 2.1% on swine farms. The prevalence in cattle in this study is lower than that reported in the United States and could be related to the lower reported prevalence of E. coli O157 in humans in Mexico. However, further research is needed to verify prevalence throughout other regions of Mexico, as well as prevalence during other seasons of the year.


Substance abuse in pregnancy.
Substance abuse in pregnancy is a major area of interest for clinicians, researchers, and public health officials. Cigarettes have long been associated with poor reproductive outcomes. Smoking increases the relative risk of placenta previa to 2.6 and of sudden infant death syndrome to 2.2 in white infants. The risk-to-benefit ratio of nicotine replacement therapy is discussed. Alcohol use by pregnant women decreased from 32% to 20% between 1985 and 1988. The long-term sequelae of fetal alcohol syndrome in adolescents and adults are presented. Marijuana is probably the most common illicit drug used in pregnancy. Long-term follow-up showed no difference in outcome until 48 months. However, at 48 months, memory and verbal measures were affected by heavy prenatal marijuana use. Self-report of illicit drug use is inaccurate. Routine urine screening for cocaine can show marked variations by locality. Analysis of meconium, urine, and infant and maternal hair along with self-report provides the most accurate information on prenatal cocaine use. The increase in cost of caring for cocaine-exposed infants was estimated. Newborns reveal decreased habituation and increased neonatal stress measures. Long-term sequelae of prenatal cocaine use indicated that smaller head circumference persist and that head size is associated with Bayley mental developmental indexes. However, the Bayley scale measured no difference among the cocaine-exposed, alcohol-exposed, or control groups.


The Closed-Can Exhalation Method for Measuring Radon.
Results from closed-can radon exhalation experiments must be interpreted bearing the time-dependent radon diffusion theory in mind. A rapid change from the free to final steady-state exhalation rate will take place for all samples that are thin compared with the radon diffusion length. The radon gas accumulating in a closed can corresponds to a free exhalation rate only if the outer volume of air is much larger than the pore volume of the enclosed sample, or the thickness of the sample is much larger than the radon diffusion length.


Analysis of Difference in Microbial Community and Physicochemical Indices Between Surface and Central Parts of Chinese Special-Flavor Baijiu Daqu.
Special-flavor Baijiu is a unique Baijiu in Jiangxi Province, China, whose uniqueness mainly depends on the unique production process of special-flavor Baijiu Daqu. However, the microbial structure and physicochemical indices of different parts of the special-flavor Baijiu Daqu are still unknown. This greatly reduces the actual value of Daqu in the production of special-flavor Baijiu. Therefore, culture-dependent and Illumina MiSeq sequencing methods were used to analyze the microbial structure of special-flavor Baijiu Daqu. The results indicated that there was a complicated microbial diversity in Chinese special-flavor Baijiu Daqu. The predominant bacterial communities were Bacillales, Lactobacillales, and Rhodospirillales, while Saccharomycetales and Eurotiales were the predominant fungal communities. Significant differences in microbial community and distribution were shown between the surface and central parts of Daqu. Acetobacter and Pichia genera were the predominant microorganisms in the surface part of Daqu, whereas Aspergillus, Kroppenstedtia, Oceanobacillus, and Bacillus genera were the predominant microorganisms in the central part of Daqu. Meantime, the different microbial distributions between the surface and central parts of Daqu caused the significant differences in the physicochemical indices. These results can provide an important theoretical basis for improving the brewing process and the quality of special-flavor Baijiu.


[Fall risk and fracture. Associated factors for falls in patients with inflammatory polyarthritis].
Patients with rheumatoid arthritis (RA) have an increased risk of falls compared to healthy controls. Previous studies reported that the incidence of fractures in RA patients was mainly caused by falls. Some factors including tender and/or swollen joint counts, low levels of physical activity and history of falls are reported to be associated with falls in the patients with RA. Control of disease activity in the patients is essential to reduce the risk of falls and the burden of fracture. Moreover, potential interventions to prevent falls such as osteoporosis therapy and balance training are necessary for the patients.


Photoinduced free radicals from chlorpromazine and related phenothiazines: relationship to phenothiazine-induced photosensitization.
Chlorpromazine and several other related phenothiazines are known to cause both phototoxic and photoallergic reactions in the skin and eyes of patients receiving these drugs. While the detailed mechanisms of photosensitization are not known, it is obvious that the first step must be the absorption of light by the drug, its metabolites, or photoproducts, or possibly an induced endogenous chemical. In this review, the free-radical photochemistry of phenothiazines is described, and the evidence for the involvement of photoinduced free radicals in photosensitization is examined. Upon irradiation chlorpromazine yields a variety of free radicals including the corresponding cation radical (via photoionization), the neutral promazinyl radical and a chlorine atom (Cl.) (via homolytic cleavage), and a sulfur-centered peroxy radical. The chlorpromazine cation radical is probably responsible for some of the observed in vitro phototoxic effects of this drug. However, it seems unlikely that the cation radical is involved in phototoxicity in vivo, since photoionization only occurs when chlorpromazine is excited into the S2 level (lambda ex less than 280 nm). The promazinyl radical is a more likely candidate for the phototoxic species both in vivo and in vitro. In addition, this radical can react covalently with proteins and other macromolecules to yield antigens which could be responsible for the photoallergic response to chlorpromazine. Neither oxygen-derived radicals nor singlet oxygen (1O2*), appear to be important in chlorpromazine photosensitization. In contrast, it would seem that promazine-induced phototoxicity may result in part from the generation of superoxide (O2-.).(ABSTRACT TRUNCATED AT 250 WORDS)


Using fluorescent chemokine uptake to detect chemokine receptors by fluorescent activated cell sorting.
Fluorescent activated cell sorting (FACS) is a powerful technique that allows rapid quantitative cell-by-cell analysis of the expression of multiple distinct proteins and the isolation of specific cell types from complex cellular mixtures. FACS-based detection of chemokine receptors is typically achieved using chemokine receptor-specific antibodies. However, this approach has some limitations, and, in our experience, many anti-chemokine receptor antibodies have poor signal-to-noise ratios and thus low sensitivity and reliability in FACS. Moreover, antibodies against some chemokine receptors are not commercially available, particularly in nonhuman species, and generating good antibodies against chemokine receptors is challenging and expensive. Here, we describe a simple alternative method of detecting chemokine receptors by FACS that relies on cellular internalization of fluorescently labelled chemokines. This approach detects chemokine receptors with high sensitivity, specificity, and reliability, and consistently outperforms antibody-based detection methods when applied to the analysis of mouse cells in our experience. It can also be used to study receptor specificity on primary cells and, since chemokines from one species often bind to receptors from other species, a single fluorescently labelled chemokine should allow reliable chemokine receptor detection and analysis across a range of species.


Self-reported Measures of Strength and Sport-Specific Skills Distinguish Ranking in an International Online Fitness Competition.
Serafini, PR, Feito, Y, and Mangine, GT. Self-reported measures of strength and sport-specific skills distinguish ranking in an international online fitness competition. J Strength Cond Res 32(12): 3483-3493, 2018-To determine if self-reported performance measures could distinguish ranking during the 2016 CrossFit Open, data from 3,000 male (n = 1,500; 27.2 ± 8.4 years; 85.2 ± 7.9 kg; 177.0 ± 6.5 cm) and women (n = 1500, 28.7 ± 4.9 years; 63.7 ± 5.8 kg; 163.7 ± 6.6 cm) competitors was used for this study. Competitors were split by gender and grouped into quintiles (Q1-Q5) based upon their final ranking. Quintiles were compared for one-repetition maximum (1RM) squat, deadlift, clean and jerk (CJ), snatch, 400 m sprint, 5,000 m run, and benchmark workouts (Fran, Helen, Grace, Filthy-50, and Fight-Gone-Bad). Separate one-way analyses of variance revealed that all competitors in Q1 reported greater (p ≤ 0.05) 1RM loads for squat (men: 201.6 ± 19.1 kg; women: 126.1 ± 13.0 kg), deadlift (men: 232.4 ± 20.5 kg; women: 148.3 ± 14.5 kg), CJ (men: 148.9 ± 12.1 kg; women: 95.7 ± 8.4 kg), and snatch (men: 119.4 ± 10.9 kg; women 76.5 ± 7.6 kg) compared with other quintiles. In addition, men in Q1 (59.3 ± 5.9 seconds) reported faster (p ≤ 0.05) 400 m times than Q3 only (62.6 ± 7.3 seconds), but were not different from any group in the 5,000 m run. Women in Q2 (67.5 ± 8.8 seconds) reported faster (p ≤ 0.05) 400 m times than Q3-Q5 (73.5-74.8 seconds), and faster (21.3 ± 1.8 minutes, p < 0.02) 5,000 m times than Q4 (22.6 ± 2.2 minutes) and Q5 (22.6 ± 1.9 minutes). Faster (p ≤ 0.05) Fran times were reported by Q1 (men: 138.2 ± 13.3 seconds; women: 159.4 ± 28.3 seconds) compared with other groups, while the results of other workouts were variable. These data indicate that the most successful athletes excel in all areas of fitness/skill, while lower-ranking athletes should focus on developing strength and power after achieving sufficient proficiency in sport-specific skills.


Intracranial extraskeletal mesenchymal chondrosarcoma.
Intracranial extraskeletal mesenchymal chondrosarcoma is a rare malignant variant of chondrosarcomas. We present a 5-year-old Saudi male child who was brought to the Emergency Department with the complaints of headache, irritability, vomiting, and unsteadiness of gait with right hemiparesis. Radiological studies confirmed the presence of a space-occupying lesion in the left cerebral hemisphere of the brain. Through a left temporoparietal craniotomy a total macroscopic excision of the tumor was carried out, and the tumor was found attached to the dura at the base of the temporal fossa. The tumor was well circumscribed, extra axial in location, and was easily dissected from the other part of the brain. The histopathological examination revealed the tumor to be a mesenchymal chondrosarcoma. This rare tumor of the pediatric age group should be included in the differential diagnosis of all intracranial tumors with aggressive characteristics.


Clinician- and Patient-reported Endpoints in CNS Orphan Drug Clinical Trials: ISCTM Position Paper on Best Practices for Endpoint Selection, Validation, Training, and Standardization.
The International Society of CNS Clinical Trials Methodology (ISCTM) Working Group on Rare Disease/Orphan Drug Development is dedicated to improving and streamlining trials to best develop new treatments for rare diseases. The rarity of these disorders requires a drug development strategy that differs from those of nonrare conditions. Rare disease drug development programs are challenged with small sample sizes, heterogeneous clinical presentations, and few, if any, off-the-shelf endpoints. When disease-specific clinical endpoints exist, they might not be validated and are typically not well known or broadly used in clinical practice. This paper aims to provide an overview of the special issues surrounding endpoints in rare disease drug development, with guidance, practical applications, and discussion. The paper covers regulatory considerations in endpoint selection; identification of relevant measurement domains; methods of quantifying clinical meaningfulness; incorporation of patient- and clinician-reported outcomes; considerations for global clinician- and patient-rated clinical assessments; cognition assessment challenges in rare diseases; translation considerations; training, standardization, and calibration of assessors; and endpoint quality assurance. Additionally, it provides guidance and resources for those involved in drug development for rare diseases. In keeping with the mission of ISCTM and the rare disease/orphan drug development working group, this article is designed to encourage thoughtful consideration and provide insight and guidance to promote and further efforts in in central nervous system (CNS) rare disease drug development efforts.


Efficacy and safety of the new TightRail™ mechanical sheath for transvenous lead extraction: Results of a French multicenter study.
The aim of this study was to assess the safety and efficacy of the TightRail™ sheath for pacemaker/defibrillator transvenous lead extraction (TLE). Multicenter observational study including patients who underwent a TLE with the TightRail™ sheath in five French university hospitals from September 2014 to January 2020. Two hundred and twenty-five patients (76% males, 71 ± 12 years) underwent a TLE procedure with the TightRail™. A total of 438 leads were extracted using the TightRail™, and the mean age of the extracted leads was 128 ± 85 months; of these, 344 (79%) were pacing leads and 94 (21%) were implantable cardioverter defibrillator leads. The overall clinical success of the extraction procedures was 93%. Overall, 410 of the 438 leads (95%) were extracted (complete or incomplete removal). After multilevel mixed-effects logistic regression model, we found that lead age (odds ratio [OR], 95% confidence interval [CI] for a 1 year increase: 1.11 [1.07-1.15], p < .001) and number of leads extracted (OR, 95% CI: 2.09 [1.50-2.96], p < .001) were the two independent factors associated with complete lead removal failure. Finally, there were 7 (3%) cases of major complications but no per-procedural death. This is the first large-scale survey assessing the efficacy and safety profile of the Tightrail™ mechanical sheath. The clinical success rate was 93%, and the lead removal failure was dependent on the age and number of leads. We show a satisfactory safety profile in this cohort of patients from primarily low-volume centers with older leads.


Utility of osteosclerotic lesion biopsy in diagnosis of POEMS syndrome: A case report.
We report a case of successful diagnosis of POEMS (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes) syndrome based on monoclonality that was confirmed by an osteosclerotic lesion biopsy in a patient without pathognomonic symptoms or monoclonal gammopathy, probably because of comorbidities, which included systemic lupus erythematosus, rheumatoid arthritis, and Sjögren syndrome. A 57-year-old woman presented with an approximately 2-year history of numbness in the toes that had gradually spread, along with muscle weakness in both arms and legs. She had been receiving immunosuppressant and corticosteroid therapy since being diagnosed with systemic lupus erythematosus and Sjögren syndrome at the age of 31 years and rheumatoid arthritis at the age of 44 years. Neurological examination revealed predominantly distal hypoesthesia and weakness in a typical stocking-and-glove pattern. Immunoelectrophoresis revealed elevated polyclonal immunoglobulin, which was attributed to her known underlying disease. Biopsy of an osteosclerotic lesion confirmed proliferation of monoclonal plasma cells, leading to a diagnosis of POEMS syndrome. Lenalidomide therapy was started after the diagnosis and the patient had a favorable outcome. Osteosclerotic lesion biopsy can be useful for diagnosis of POEMS syndrome in difficult cases.


Optimizing sensitization processes in dinuclear luminescent lanthanide oligomers: selection of rigid aromatic spacers.
This work illustrates a simple approach for optimizing the lanthanide luminescence in molecular dinuclear lanthanide complexes and identifies a particular multidentate europium complex as the best candidate for further incorporation into polymeric materials. The central phenyl ring in the bis-tridentate model ligands L3–L5, which are substituted with neutral (X = H, L3), electron-withdrawing (X = F, L4), or electron-donating (X = OCH3, L5) groups, separates the 2,6-bis(benzimidazol-2-yl)pyridine binding units of linear oligomeric multi-tridentate ligand strands that are designed for the complexation of luminescent trivalent lanthanides, Ln(III). Reactions of L3–L5 with [Ln(hfac)3(diglyme)] (hfac– is the hexafluoroacetylacetonate anion) produce saturated single-stranded dumbbell-shaped complexes [Ln2(Lk)(hfac)6] (k = 3–5), in which the lanthanide ions of the two nine-coordinate neutral [N3Ln(hfac)3] units are separated by 12–14 Å. The thermodynamic affinities of [Ln(hfac)3] for the tridentate binding sites in L3–L5 are average (6.6 ≤ log(β(2,1)(Y,Lk)) ≤ 8.4) but still result in 15–30% dissociation at millimolar concentrations in acetonitrile. In addition to the empirical solubility trend found in organic solvents (L4 > L3 >> L5), which suggests that the 1,4-difluorophenyl spacer in L4 is preferable, we have developed a novel tool for deciphering the photophysical sensitization processes operating in [Eu2(Lk)(hfac)6]. A simple interpretation of the complete set of rate constants characterizing the energy migration mechanisms provides straightforward objective criteria for the selection of [Eu2(L4)(hfac)6] as the most promising building block.


[Animal models for retinitis pigmentosa research].
Retinitis pigmentosa (RP) is the general term given to a group of genetically determined, degenerative retinal diseases which afflict some 1.5 million humans worldwide. Molecular genetic studies in recent years have shown that RP cannot be explained by a single genetic defect but rather that the hereditary aberration responsible for triggering the onset of the disease is localized in different genes and at different sites within these genes. A fuller understanding of these processes is possible only if the pathogenesis of the diseases can be followed as they develop. Animal models are an indispensable requirement for this. Only in the animal model is it possible to observe progressive degenerations in "time-lapse photography" as it were and to examine the biochemical, functional and morphological changes which appear at specific times in the retina. This article gives an overview of the animal models presently used in RP research, with a short description of genetic backgrounds and the progression of the disease in individual animals. In addition, a look at the future is provided of new molecular techniques which make it possible to create specific animal models with gene mutations identical to those appearing in human RP. Since hereditary human retinal degenerations can be brought about by numerous different genetic defects, comparative studies using animals with clearly defined genetic defects are essential. Only in this way is it possible to arrive at conclusions concerning this specialized syndrome and to clarify the mechanisms which are common to all retinal degenerations.


Toward a Molecular Classification of Colorectal Cancer: The Role of MGMT.
O(6)-methylguanine DNA methyltransferase (MGMT) is a DNA repair enzyme with the ability to protect cells from DNA mutations by removing alkyl groups from the O(6) position of guanine. Colon mucosa is exposed to the direct effects of environmental carcinogens and therefore maintaining a proficient DNA repair system is very important to stay protected against DNA mutagenesis. Loss of MGMT expression is almost exclusively associated with methylation of CpG islands in the MGMT gene promoter region which is found in approximately 40% of colorectal cancers. The role of MGMT loss in colorectal tumorigenesis is complex but numerous studies have documented methylation of this gene even in the normal appearing mucosa as well as in aberrant crypt foci, suggesting that MGMT methylation can be regarded as an early event or "field defect" in colon cancer neoplasia. The focus of this perspective is the role of MGMT in different pathways of colorectal carcinogenesis as well as the implication of this molecule in treatment decisions in colorectal cancer patients.


The incidence, clinical characteristics, and outcome of COVID-19 in a prospectively followed cohort of patients with Behçet's syndrome.
Initial case series of small number of patients at the beginning of the pandemic reported a rather guarded prognosis for Behçet's syndrome (BS) patients infected with SARS-CoV-2. In this prospective study, we describe the incidence, clinical characteristics, disease course, management, and outcome in a large cohort of BS patients with laboratory-confirmed infection of SARS-CoV-2. We defined a cohort of 1047 registered BS patients who were aged between 16 and 60 years and seen routinely before the pandemic at the multidisciplinary outpatient clinic. We followed prospectively this cohort from beginning of April 2020 until the end of April 2021. During 13 months of follow-up, of the 1047 (599 M/448 F) patients, 592 (56.5%) were tested for SARS-CoV-2 PCR at least once and 215 (20.5%; 95% CI 0.18-0.23) were tested positive. We observed 2 peaks which took place in December 2020 and April 2021. Of the 215 PCR positive patients, complete information was available in 214. Of these 214, 14 (6.5%) were asymptomatic for COVID-19. In the remaining, the most common symptoms were anosmia, fatigue, fever, arthralgia, and headache. A total of 40 (18.7%) had lung involvement, 25 (11.7%) were hospitalized, 1 was admitted to the intensive care unit while none died. Favipiravir was the most prescribed drug (74.3%), followed by colchicine (40.2%), and hydroxychloroquine (20.1%) in the treatment of COVID-19. After COVID-19, 5 patients (2.3%) were given supplemental O2 and 31 (14.5%) antiaggregant or anticoagulants. During COVID-19, of the 214 PCR positive patients, 116 (54.2%) decreased the dose of their immunosuppressives or stopped taking completely; 36 (16.8%) experienced a BS flare which was mostly oral ulcers (10.3%). None of the patients reported a thrombotic event. A total of 93 (43.5%) patients reported BS flares after a median 45 days of COVID-19 infection and this was found to be significantly associated with immunosuppressive drug discontinuation. Multiple regression analysis adjusted for age and gender indicated that smoking and using interferon-alpha decreased the likelihood of getting COVID-19. The incidence and severity of COVID-19 did not differ between those who were using colchicine or not. The cumulative incidence of COVID-19 in this prospectively followed cohort of BS patients was almost two folds of that estimated for the general population living in Istanbul, Turkey, however, the clinical outcome of COVID-19 was not severe and there was no mortality. The protective effect of smoking and interferon deserves further investigation. On the other hand, colchicine did not have any positive or negative effect against COVID-19. Significant number of patients flared after COVID-19, however, this was significantly associated with immunosuppressive discontinuation during the infection. Contrary to our previous observations, COVID-19 did not seem to exacerbate thrombotic events during or after the infection.


Review of Sepsis in Burn Patients in 2020.
Background: Severe burn injury results in substantial damage to the skin, inhibiting its ability to perform as the primary barrier to infection. Additionally, severe burn injury can lead to critical illness and extensive time in the intensive care unit (ICU). These two factors work to increase the risk of sepsis in the burn patient compared with other hospitalized patients. The increased risk of sepsis is compounded by the difficulty of diagnosing sepsis in severely burned patients because the pathophysiology of large burns mimics sepsis, leading to possible delay in diagnosis and initiation of treatment. Methods: A literature review was performed to discuss and review the diagnostic difficulties and criteria used to identify patients with sepsis. Additionally, the most current management of sepsis was reviewed and described in caring for burn patients with sepsis. Results: The incidence of sepsis in patients with more than 20% total body surface area (TBSA) burns is between 3% and 30% and is the most common cause of death in the burn patient, with pneumonia being the most common etiology. Several different diagnostic criteria for diagnosing sepsis in burn patients exist, however, none of these criteria have proven to be superior to clinical diagnosis by an experienced burn surgeon. As with sepsis in other patient populations, prompt diagnosis, initiation of antibiotic agents, and source control remain the standard management of sepsis in the burn patient. Conclusions: Because of the loss of the primary infection barrier function of the skin after a substantial burn injury, this patient population is at increased risk for sepsis. Because of the pathophysiology of burn injuries, diagnosing sepsis in the burn population remains challenging. Understanding the most common etiologies of sepsis in burn patients may help with more expedient diagnosis and initiation of treatment.


Chemotaxis-like regulatory systems: unique roles in diverse bacteria.
Bacteria sense the chemical world using a variety of mechanisms that include the frequently described two-component system (TCS), which comprises a sensor kinase and response regulator, to regulate gene expression in response to environmental cues. One of the best and most widely studied versions of the TCS is the system that controls chemotaxis in Escherichia coli. The chemotaxis machinery includes components not found in other TCS to regulate motility and is therefore an exception to the rule for two-component signaling. The hallmark feature of the chemotaxis system is the presence of an adaptation module in which the sensor receptor protein is posttranslationally modified to attenuate ligand-induced signaling, a mechanism not yet identified for the more widely distributed prototypical TCS. More recently, variations on the chemotaxis system itself have been identified and they are termed chemosensory systems and are the subject of this review. Extensive research has provided a perspective on TCS signaling and indicates that variation and diversity for the standard two-component system are predominant in the microbial world.


Semester-long inquiry-based molecular biology laboratory: Transcriptional regulation in yeast.
A single semester molecular biology laboratory has been developed in which students design and execute a project examining transcriptional regulation in Saccharomyces cerevisiae. Three weeks of planning are allocated to developing a hypothesis through literature searches and use of bioinformatics. Common experimental plans address a cell process and how three genes that encode for proteins involved in that process are transcriptionally regulated in response to changing environmental conditions. Planning includes designing oligonucleotides to amplify the putative promoters of the three genes of interest. After the PCR, each product is cloned proximal to β-galactosidase in a yeast reporter plasmid. Techniques used include agarose electrophoresis, extraction of DNA from agarose, plasmid purification from bacteria, restriction digestion, ligation, and bacterial transformation. This promoter/reporter plasmid is then transformed into yeast. Transformed yeast are cultured in conditions prescribed in the experimental design, lysed and β-galactosidase activity is measured. The course provides an independent research experience in a group setting. Notebooks are maintained on-line with regular feedback. Projects culminate with the presentation of a poster worth 60% of the grade. Over the last three years, about 65% of students met expectations for experimental design, data acquisition, and analysis. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(2):145-151, 2017.


Effects of Different Ligands in the Notch Signaling Pathway on the Proliferation and Transdifferentiation of Primary Type II Alveolar Epithelial Cells.
Background: Transdifferentiation of type II alveolar epithelial cells (AECII) into type I alveolar epithelial cells (AECI) is involved in neonatal respiratory distress syndrome (NRDS). Different ligands of the Notch pathway could have different effects on AECII transdifferentiation. Objective: To investigate the effects of Dlk1 and Jagged1 on the proliferation and transdifferentiation of AECII. Methods: Fetal AECIIs (19 days of gestation) were divided: control group, Dlk1 group, rhNF-κB group. Proliferation was tested using the MTT assay. Expression of surfactant protein C (SP-C) and aquaporin 5 (AQP5) was examined by immunofluorescence. mRNA and protein levels of SP-C, AQP5, Nortch1, Dlk1, Jagged1, and Hes1 were examined by RT-PCR and western blot. Results: In response to Dlk1, cell number and proliferation were increased (P < 0.05), and mRNA and protein levels of SP-C, Dlk1, Notch1, and Hes1 were up-regulated, while AQP and Jagged1 were decreased. In response to rhNF-κB, the cell number and proliferation were reduced, and mRNA and protein levels of Jagged1 and Notch1 were up-regulated, while Dlk1, and SP-C were downregulated. In the Dlk1 group, SP-C, and AQP5 expression patterns suggested that the cells were still transdifferentiating by 96 h, while in the rhNF-κB group, most cells had transdifferentiated by 72 h and were close to apoptosis by 96 h. Conclusion: These results suggest that Dlk1 promoted proliferation of AECIIs and inhibited cell transdifferentiation, while Jagged1 treatment inhibited proliferation of AECIIs and promoted transdifferentiation to AECIs. These results provide some clue for the eventual management of NDRS.


DOT-1.1-dependent H3K79 methylation promotes normal meiotic progression and meiotic checkpoint function in C. elegans.
Epigenetic modifiers are emerging as important regulators of the genome. However, how they regulate specific processes during meiosis is not well understood. Methylation of H3K79 by the histone methyltransferase Dot1 has been shown to be involved in the maintenance of genomic stability in various organisms. In S. cerevisiae, Dot1 modulates the meiotic checkpoint response triggered by synapsis and/or recombination defects by promoting Hop1-dependent Mek1 activation and Hop1 distribution along unsynapsed meiotic chromosomes, at least in part, by regulating Pch2 localization. However, how this protein regulates meiosis in metazoans is unknown. Here, we describe the effects of H3K79me depletion via analysis of dot-1.1 or zfp-1 mutants during meiosis in Caenorhabditis elegans. We observed decreased fertility and increased embryonic lethality in dot-1.1 mutants suggesting meiotic dysfunction. We show that DOT-1.1 plays a role in the regulation of pairing, synapsis and recombination in the worm. Furthermore, we demonstrate that DOT-1.1 is an important regulator of mechanisms surveilling chromosome synapsis during meiosis. In sum, our results reveal that regulation of H3K79me plays an important role in coordinating events during meiosis in C. elegans.


The Link between the Marital Bond and Future Triadic Family Interactions.
This study examined how the marital bond, as indexed through the Oral History Interview (OHI), is related to future triadic family interactions. Families (N = 108), with a 7 - 9 year old child, participated in a longitudinal study (the Family Health Project) examining children's emotional development throughout the transition to adolescence. Parental cohesion and family cohesion, warmth, structure, and problem solving were assessed via behavioral observation during family problem solving discussions and parent-child teaching interactions 18 - 24 months after the OHI. Results indicated that the marital bond was predictive of parental cohesion, family cohesion, warmth, and structure during teaching interactions. The marital bond was not significantly predictive of family problem solving or parental cohesion in problem solving interactions.


Between a Rock and a Hard Polytomy: Phylogenomics of the Rock-Dwelling Mbuna Cichlids of Lake Malaŵi.
Whole genome sequences are beginning to revolutionize our understanding of phylogenetic relationships. Yet, even whole genome sequences can fail to resolve the evolutionary history of the most rapidly radiating lineages, where incomplete lineage sorting, standing genetic variation, introgression, and other factors obscure the phylogenetic history of the group. To overcome such challenges, one emerging strategy is to integrate results across different methods. Most such approaches have been implemented on reduced representation genomic data sets, but whole genomes should provide the maximum possible evidence approach. Here, we test the ability of single nucleotide polymorphisms extracted from whole genome resequencing data, implemented in an integrative genomic approach, to resolve key nodes in the phylogeny of the mbuna, rock-dwelling cichlid fishes of Lake Malaŵi, which epitomize the phylogenetic intractability that often accompanies explosive lineage diversification. This monophyletic radiation has diversified at an unparalleled rate into several hundred species in less than 2 million years. Using an array of phylogenomic methods, we consistently recovered four major clades of mbuna, but a large basal polytomy among them. Although introgression between clades apparently contributed to the challenge of phylogenetic reconstruction, reduction of the data set to nonintrogressed sites still did not help to resolve the basal polytomy. On the other hand, relationships among six congeneric species pairs were resolved without ambiguity, even in one case where existing data led us to predict that resolution would be difficult. We conclude that the bursts of diversification at the earliest stages of the mbuna radiation may be phylogenetically unresolvable, but other regions of the tree are phylogenetically clearly supported. Integration of multiple phylogenomic approaches will continue to increase confidence in relationships inferred from these and other whole-genome data sets. [Incomplete lineage sorting; introgression; linkage disequilibrium; multispecies coalescence; rapid radiation; soft polytomy.].


On the optimization principle in phylogenetic analysis and the minimum-evolution criterion.
This paper discusses the optimization principle in phylogenetic analysis, in the case of distance data. We argue that the use of this principle cannot be called into question, except for computing time reasons. We show that the minimum-evolution criterion is not perfectly suited for distance data estimated from sequences, and we present another approach, implemented in the BIONJ algorithm, which allows the data features to be taken into account, while being less demanding in computing time. Simulations show that BIONJ significantly outperforms NJ.


Learning-induced biases in the ongoing dynamics of sensory representations predict stimulus generalization.
Sensory stimuli have long been thought to be represented in the brain as activity patterns of specific neuronal assemblies. However, we still know relatively little about the long-term dynamics of sensory representations. Using chronic in vivo calcium imaging in the mouse auditory cortex, we find that sensory representations undergo continuous recombination, even under behaviorally stable conditions. Auditory cued fear conditioning introduces a bias into these ongoing dynamics, resulting in a long-lasting increase in the number of stimuli activating the same subset of neurons. This plasticity is specific for stimuli sharing representational similarity to the conditioned sound prior to conditioning and predicts behaviorally observed stimulus generalization. Our findings demonstrate that learning-induced plasticity leading to a representational linkage between the conditioned stimulus and non-conditioned stimuli weaves into ongoing dynamics of the brain rather than acting on an otherwise static substrate.


Medical and surgical devices in the emergency and trauma patient: what the radiologist should know, and how they can add value.
A variety of different external and internal medical devices are used in the acute setting to maintain life support and manage severely injured and unstable trauma or emergency patients. These devices are inserted into the acutely ill patient with the specific purpose of improving outcome, but misplacement can cause additional morbidity and mortality. Consequently, meaningful interpretation of the position of devices can affect acute management. Some devices such as nasopharyngeal, nasogastric and endotracheal tubes and chest and surgical drains are well known to most clinicians, however, little formal training exists for radiologists in composing their report on the imaging of these devices. The novice radiologist often relies on tips and phrases handed down in an aural tradition or resorts to phrases such as: "position as shown". Furthermore, radiologists with limited experience in trauma might not be familiar with the radiological appearance of other more specific devices. This review will focus on the most common medical devices used in acute trauma patients, indications, radiological appearance and their correct and suboptimal positioning.


Postoperative changes at the lower end of cervical laminoplasty: for preservation of the C7 spinous process in laminoplasty.
C3-C7 laminoplasty has been the standard treatment for cervical myelopathy, although several recent reports described C3-C6 laminoplasty for preserving the muscles inserting in C7 and reducing postoperative axial symptoms. However, postoperative changes at C6/C7 of the lower end of C3-C6 laminoplasty, especially regarding a possibility of postoperative spinal canal narrowing have not been measured. The purpose of this study was to clarify postoperative changes at the lower end of laminoplasty. Pre and postoperative spinal dura diameter at the lower end of conventional C3-C7 laminoplasty using MRI, and the related factors for spinal dura diameter and the causes of postoperative dura narrowing were investigated. At the last follow-up after C3-C7 laminoplasty, dura diameter at C7/T1 was significantly wider after operation than before operation, and postoperative narrowing of dura diameter, which was found in 20% of patients, was a maximum amount of one millimeter. No pre and postoperative factor significantly correlated with dura diameter at C7/T1. The causes of postoperative narrowing at the lower end of laminoplasty were disc protrusion and/or posterior scar, or segmental angulation of the spinal cord. In conclusion, the presence of preoperative subarachnoid space over one millimeter at C6/C7 may be able to be one of the radiological indications for C3-C6 laminoplasty.


Radiation induces epithelial-mesenchymal transition in colorectal cancer cells.
Radiotherapy remains a major approach to adjuvant therapy for patients with advanced rectal cancer. Nevertheless, the effects of radiation on malignant processes have yet to be clarified. The aim of this study was to assess the biological effects of radiation on colorectal cancer (CRC) cells with special reference to epithelial-mesenchymal transition (EMT), a key developmental program often activated during cancer invasion and metastasis. We investigated the effect of radiation on two colorectal cancer cell lines, CaR1 and DLD1, assessing cell morphology, motility, migration and invasive ability. Expression of molecules associated with EMT was determined using RT-PCR, Western blotting, and immunofluorescence staining in control and irradiated cells. We also used real-time RT-PCR to examine the expression of molecules associated with EMT before and after chemoradiotherapy. Thus, we studied 26 rectal cancer patients who received preoperative chemoradiotherapy followed by radical surgery. In addition, we examined the relationship between disease recurrence and the expression of a number of proteins. Irradiation caused CRC cells to undergo phenotypic changes characteristic of EMT: spindle-cell shape, loss of polarity, intercellular separation and pseudopodia formation. Irradiation enhanced cell migration and invasiveness. In irradiated CRC cells, molecular changes consistent with EMT were observed. In clinical samples, we observed molecular changes consistent with EMT, and those changes were significantly enhanced in patients with recurring disease. These results indicate that irradiation induces an alteration to a malignant phenotype consistent with EMT in colorectal cancer cells.


Quality assessment by external bodies: intended and unintended impact on healthcare delivery.
External quality assessment programs in the form of pay for performance, report cards and national rankings are rapidly overtaking more traditional, internal quality assessment efforts and external clinical practice guidelines. Although such initiatives are designed to improve healthcare quality by promoting competition and increasing transparency, review of their efficacy and unintended effects is just coming to the national spotlight. Critical evaluation of external quality assessment programs remains limited despite their scope, speed and breadth of implementation. Recent publications, however, suggest that external quality assessment efforts may have major unintended consequences. These include effects on patient decision-making, the 'dynamic equilibrium' of patient care, healthcare disparities, medical innovation and practice patterns. In their early years of implementation, external quality assessment programs have already had significant consequences in the healthcare system. As new tools become available, their full impact on care and caregivers must be thoroughly evaluated. Careful consideration of clinical practice implications and an understanding of the risks are critical before accepting and implementing new assessment paradigms. The substantial and widespread effects of these programs should prompt further evaluation from the medical community.


Partial nephrectomy for renal cortical tumors: pathologic findings and impact on outcome.
To review the experience with partial nephrectomy for the diagnosis and treatment of renal cortical tumors at the Memorial Sloan-Kettering Cancer Center with a particular focus on the histologic findings at the time of surgery. The results of 292 consecutive partial nephrectomies were reviewed. Demographic data, perioperative complications, and disease status were recorded. The specimens were reviewed for histologic subtyping in accordance with the Heidelberg classification for renal cortical tumors. The probabilities of recurrence and disease-specific death were assessed with the cumulative incidence estimate. The median follow-up was 25.3 months (range 1 to 155). A total of 246 patients (87%) were diagnosed with a renal cortical tumor, 1 patient (less than 1%) was diagnosed with another cancer, and 34 (12%) were diagnosed with benign lesions. The most frequent histologic finding among the renal cortical tumor subgroup was conventional clear cell carcinoma in 148 cases (51%), followed by papillary carcinoma in 54 (18%), oncocytoma in 32 (11%), and chromophobe carcinoma in 21 (7%). Bilateral disease and multifocality were most prevalent in the conventional and papillary subtypes, respectively. The 5-year probability of recurrence and disease-specific death was 12% and 8%, respectively, for the conventional clear cell type. No local or distant disease recurrence was observed in any other renal cortical tumor subtypes. Partial nephrectomy should be considered a diagnostic and therapeutic surgical approach for renal cortical masses. Conventional clear cell carcinoma is the most frequent histologic subtype and is associated with a less favorable outcome compared with papillary carcinoma and chromophobe carcinoma. Radical nephrectomy for renal lesions that could be removed by partial nephrectomy will risk renal impairment in a substantial proportion of patients with benign disease.


Plasma glucose and pancreatic insulin content in the early phase of streptozotocin-induced diabetes (preliminary results).
The susceptibility of female and male C57Bl/KsJ-mice to multiple low-dose streptozotocin applications was investigated. Both sexes of the strains studied developed hyperglycemia. In the early phase of streptozotocin-applications we failed to observe a strong parallelism between increase of plasma glucose and decrease of pancreatic insulin content.


MDM2 does not influence p53-mediated sensitivity to DNA-damaging drugs.
MDM2 inhibits transactivation properties of the tumor suppressor protein p53 by binding to and facilitating proteasomal degradation of p53. Because MDM2 targets p53 for degradation, it was anticipated that cells that overexpress MDM2 would not contain functional wild-type p53 (wtp53). However, p53 and MDM2 in cells with damaged DNA can become phosphorylated, and their binding to each other can become inhibited. Thus, p53 remains functional and induces apoptosis of damaged cells. Here we report the results of experiments designed to investigate whether MDM2 amplification and overexpression can inhibit p53-mediated chemosensitivity to DNA-damaging drugs. Two cell lines in which MDM2 is amplified, NB-1691 and Rh18, were transduced with an adenoviral expression vector for p53 (Ad.p53). Although functional wtp53 was detected, no change in chemosensitivity was observed, suggesting that endogenous wtp53 may have been active in the MDM2-amplified cells. The adenoviral vector Ad.MDM2 was used to generate MDM2 expression in a rhabdomyosarcoma cell line, Rh30-CI.27, engineered to express inducible wtp53. When p53 expression was induced, cells became chemosensitive to actinomycin D in the presence or absence of MDM2 expression; this result suggests that MDM2 cannot inhibit p53-mediated chemosensitivity. There was no evidence of a reduced amount of MDM2-p53 binding after drug exposure, but the remaining unbound wtp53 may be functional and capable of potentiating cytotoxicity. In conclusion, MDM2 expression is important in inhibiting p53 function during tumor development but not during the DNA damage-mediated cytotoxic response.


Immunophenotypic profile of acute leukemia: critical analysis and insights gained at a tertiary care center in India.
To analyze the spectrum of various types and subtypes of acute leukemia. Two thousand five hundred and eleven consecutive new referral cases of acute leukemia (AL) were evaluated based on WHO classification. It included 1,471 cases (58%) of acute lymphoblastic leukemia (ALL), 964 cases (38%) of acute myeloid leukemia (AML), 45 cases (1.8%) of chronic myelogenous leukemia in blast crisis (CMLBC), 37 cases (1.5%) of biphenotypic acute leukemia (BAL), 1 case of Triphenotypic AL, and 2 cases of acute undifferentiated leukemia (AUL). Common subtypes of ALL were B-cell ALL (76%), which comprised of intermediate stage/CALLA positive (73%), early precursor/proBALL (3%). T-cell ALL constituted 24% (351 cases) of ALL. Common subtypes of AML included AMLM2 (27%), AMLM5 (15%), AMLM0 (12%), AMLM1 (12%), APML (11%), and AML t(8;21) (9%). CMLBC was commonly of myeloid blast crisis subtype (40 cases). B-cell ALL was the commonest subtype in children and AML in adults. Overall incidence of AML in adults was low (53% only). CD13 was most sensitive and CD117 most specific for determining myeloid lineage. A minimal primary panel of nine antibodies consisting of three myeloid markers (CD13, CD33, and CD117), B-cell lymphoid marker (CD19), T-cell marker (CD7), with CD45, CD10, CD34, and HLADR could assign lineage to 92% of AL. Cytogenetics findings lead to a change in the diagnostic subtype of myeloid malignancy in 38 (1.5%) cases.


Translational diffusion across a free-standing smectic film above the bulk smectic-A-isotropic transition temperature.
Calculations of translational self-diffusion coefficient in free-standing smectic films during a series of layer-thinning transitions as the temperature is raised above the bulk smectic-A-isotropic transition have been carried out. A molecular model based upon the random walk theory is applied for calculating the translational diffusion coefficient (TDC) D_{∥} across the smectic film both in the bulk of the film, as well as in the vicinity of the bounding surfaces. Calculations of D_{∥} require the set of the orientational and translational order parameters (OPs) which have been obtained by using the extended McMillan approach with anisotropic forces. The effect of E on the orientational and translational OPs, as well as on the TDC of smectic films has been investigated. A reasonable agreement between the theoretically predicted and the experimentally obtained data on the TDC in the bulk of the partially fluorinated H10F5MOPP film has been obtained. We also found, in agreement with the experimentally observed behavior of D_{∥}(N)(N=25,13,11,10), that the translational diffusion coefficient in the bulk of the film gradually increases as the film thickness N is decreased.


[Effect of nedocromil sodium on the hyperreactivity of the bronchial system in young asthmatic patients].
The protective effect of nedocromil sodium on asthmatic children was investigated by exercise provocation test in a placebo-controlled, double-blind, randomized crossover trial. Lung function impairment during exercise was significantly less (p less than 0.05) with nedocromil sodium than with placebo. The maximum impairment of lung function after exercise challenge was significantly less (p less than 0.05) with nedocromil sodium than with placebo. No side-effect were observed.


Plasmonic hot carrier-driven photoelectrochemical water splitting on antenna-reactor Pt/Ag/TiO2 Schottky nanodiodes.
Plasmonic photoelectrochemical (PEC) water splitting has excited immense interest, as it can overcome the intrinsic limitations of semiconductors, in terms of light absorption, by the localized-surface plasmon resonances effect. Here, to get insight into the role of plasmonic hot carriers in plasmonic water splitting, a rational design of an antenna-reactor type Pt/Ag/TiO2 metal-semiconductor Schottky nanodiode was fabricated and used as a photoanode. Using the designed PEC cell system combined with the Pt/Ag/TiO2 nanodiode, we show that the plasmonic hot carriers excited from Ag were utilized for the oxygen (O2) evolution reaction and, consequently, had a decisive role in the enhancement of the photocatalytic efficiency. These results were supported by finite-difference time-domain simulations, and the faradaic efficiency was measured by the amount of actual gas produced. Therefore, this study provides a deep understanding of the dynamics and mechanisms of plasmonic hot carriers in plasmonic-assisted PEC water splitting.


Therapeutic potential of CRF receptor antagonists: a gut-brain perspective.
Activation of the corticotropin-releasing factor (CRF) family of neuropeptide receptors in the brain and periphery appears to mediate stress-related changes in a variety of physiological and functional domains. Comparative pharmacology of CRF receptor agonists suggests that CRF, urocortin, sauvagine and urotensin consistently mimic, and conversely peptide CRF receptor antagonists lessen, the functional consequences of stressor exposure. Together with the development of novel non-peptide CRF receptor antagonists, a growing number of CRF receptor selective ligands are available to elucidate the neurobiology and physiological role of CRF systems. The present review considers available preclinical evidence as well as results from one Phase II clinical trial which address the hypothesis that CRF receptor antagonists may represent a new option for pharmacotherapy of stress-related disorders.


The Papilla Presence Index (PPI): a new system to assess interproximal papillary levels.
The present article describes a classification system used to assess the interdental papillary height on natural teeth; the index can also be used in cases of lack of contact points and presence of diastemata. This system is particularly useful to define the loss of the interdental papilla because of periodontal disease and papilla reconstruction after periodontal treatment, particularly if tooth repositioning is involved. The index, based on esthetic evaluations, measures interproximal soft tissue height in relation to the CEJ, adjacent teeth, and the point corresponding to the ideal contact point. The four index scores are useful for an easy clinical assessment with respect to patient esthetics.


Different contractile effects of alpha1- and alpha2-adrenergic agonists on horse isolated common digital artery smooth muscle ring preparations in vitro.
Despite assays on ring preparations in vitro confirmed that the vasoconstrictor sympathetic control in the horse common digital artery mainly depends on alpha(1)-adrenoceptors stimulation, selective alpha(2)-adrenoceptor agonists were investigated under the same experimental conditions. Both detomidine (DET) and UK 14304 differed from noradrenaline (NA) and phenylephrine (PHE) in provoking contractile effects which were slowly onsetting, concentrations-unrelated and unremovable by repeated washings. While prazosin (PRA) clearly antagonized the effects of NA and PHE, neither pre- nor post-treatments of the preparations with alpha(2)-antagonists succeeded in antagonizing or removing the effects of the two alpha(2)-agonists tested, which moreover were unaffectable either by lowering the organ bath temperature or by depriving the nutritive medium of Ca(2+). To explain this unusual behavior of alpha(2)-adrenoceptors stimulation it has been hypothesized that a Ca(2+) mobilization from the endoplasmic reticulum of the smooth muscle cell occurs which is followed by a hindered reuptake of them.


Early Gut Microbiota Colonisation of Premature Infants Fed with Breastmilk or Formula with or without Probiotics: A Cohort Study.
Premature infants have a fragile ecology of the gut microbiota, which is associated with many health problems and may be influenced by formula versus breast feeding. The present study investigated differences in the process of gut microbiota colonisation in preterm infants fed with breastmilk or formula with or without probiotics before 12 weeks. This cohort study recruited 138 premature infants; 31 in the breastmilk (BM) group, 59 in the probiotics formula (PF) group and 48 in the non-probiotics formula (NPF) group, according to the feeding practice they received at birth. Gut bacterial composition was identified with 16S rRNA gene sequencing in faecal samples collected at 1 week, 6 weeks and 12 weeks after birth. The alpha diversity was higher in the PF group compared to the other groups at week 1 and 6 (both p < 0.01) but showed no difference at week 12. The beta diversity of the three groups showed a trend towards similarity at the first two stages (p < 0.001 and p = 0.009, respectively) and finally showed no difference at week 12. Canonical redundancy analysis showed that feeding type could explain the difference in gut microbiota composition at week one and six (both p < 0.01). At genus level, Bifidobacterium was enriched in the PF group, while the Enterococcus and Streptococcus was enriched in the NPF group. In summary, formula with probiotics feeding after birth can affect gut microbiota colonisation and lead to a bacterial community with less potential pathogens.


Compression Stocking Compliance Does Not Impact Reintervention or Symptom Change after Iliac Vein Stenting in Patients with Moderate to Severe Lower Extremity Edema.
Medical compression stocking (MCS) therapy remains a core treatment for chronic venous disease, particularly in patients with lower extremity edema. This study investigated the impact of postoperative MCS adherence on patients with moderate to severe edema who underwent iliac vein stenting (IVS). Patients at a large tertiary care center who underwent IVS from August 2011 to June 2021 were analyzed. Symptoms were quantified using the venous clinical severity score (VCSS) and clinical assessment score (CAS). The criterion for inclusion was moderate or severe lower extremity edema at the time of IVS. Patients were stratified into two cohorts: complete postoperative compression therapy compliance (MCS+) and no postoperative compression therapy use (MCS-). Among 376 patients fitting our study criteria, we identified 168 MCS+ and 208 MCS- patients. The VCSS edema score did not significantly differ between groups (P = 0.179). Postoperatively, the mean changes in VCSS edema at the first postoperative visit, the one-year follow-up, and the two-year follow-up were not significantly different between the MCS+ and MCS- cohorts (P = 0.123, 0.296, and 0.534, respectively). An analysis of CAS for edema revealed that the MCS+ cohort had a modestly better improvement in edema at the 90-day follow-up visit versus the MCS- cohort (P = 0.018), but this difference was not observed in the 30-day (P = 0.834) or six-month follow-up visit (P = 0.755). A multivariate analysis revealed no difference in the need for major intervention (OR 0.93, 95% CI 0.44 to 1.50, P = 0.504). A Kaplan-Meier analysis via log-rank test revealed no difference in reintervention-free survival between groups (P = 0.77). Many patients with moderate to severe lower extremity edema experience a reduction in their edema after IVS. In our study, compression stocking compliance after surgery had a little impact on edema relief in this population.


Overlapping loss of heterozygosity by mitotic recombination on mouse chromosome 7F1-ter in skin carcinogenesis.
A significant role for mouse chromosome 7 abnormalities during chemically induced skin carcinogenesis has been advanced based on previous cytogenetic and molecular studies. To determine the frequency of allelic losses at different loci of chromosome 7 in skin tumors induced in the outbred SENCAR mouse stock by a two-stage initiation-promotion protocol, we compared the constitutional and tumor genotypes of premalignant papillomas and squamous cell carcinomas for loss of heterozygosity at different informative loci. In a previous study, these tumors had been analyzed for their allelic composition at the Harvey ras-1 (Ha-ras-1) locus and it was found that 39% of squamous cell carcinomas had lost the normal Ha-ras-1 allele exhibiting 3 or 2 copies of the mutated counterpart or gene amplification. In the present study, by combining Southern blot and polymerase chain reaction fragment length polymorphism analyses, we detected complete loss of heterozygosity at the beta-globin (Hbb) locus, distal to Ha-ras-1, in 15 of 20 (75%) skin carcinomas. In addition, 5 of 5 informative cases attained homozygosity at the int-2 locus, 27 centimorgans distal to Hbb. Polymerase chain reaction analysis of DNA extracted from papillomas devoid of stromal contamination by fluorescence-activated sorting of single cell dispersions immunolabeled with anti-keratin 13 antibody revealed loss of heterozygosity at the Hbb locus, demonstrating that this event occurs during premalignant stages of tumor development. Interestingly, loss of heterozygosity was only detected in late-stage lesions exhibiting a high degree of dysplasia and areas of microinvasion. Analysis of allelic ratios by densitometric scanning of tumors that had become homozygous at Hbb but retained heterozygosis at Ha-ras-1 indicated mitotic recombination as the mechanism underlying loss of heterozygosity on mouse chromosome 7 during chemically induced skin carcinogenesis. These findings are consistent with the presence of a putative tumor suppressor gene linked to the Hbb locus in the 7F1-ter region of mouse chromosome 7, the functional inactivation of which may constitute a critical event in skin tumor progression, possibly during the malignant conversion stage.


Challenges and opportunities for treatment and research of AIDS-related malignancies in Africa.
Following our review of AIDS-associated cancer in developing nations in 2004, we sought to update recent publications and review data on the challenges and opportunities for the treatment and research of AIDS malignancies in Africa. It is apparent that the burden of AIDS-related malignancies and other virus-associated tumors is significant and increasing in Africa. Several recent studies report findings on conjunctival squamous cell carcinoma and there is a report that Hodgkin's disease, a non-AIDS-defining neoplasm, is increasing in incidence. International collaborative partnerships dedicated to AIDS malignancies in developing countries are feasible and invaluable for clinical strategies to address this aspect of the pandemic. A departure point is the ongoing work of the East Africa - Case Western Reserve University Collaboration in AIDS malignancies. The burden of neoplastic complications of HIV infection and endemic virus-associated tumors are assuming increasing significance in Africa. There is a need to develop nonmyelotoxic therapies and approaches that are hypothesis-driven and pathogenesis-based. The scarcity and shortages in this region demand that our scientific and therapeutic strategies are both suitable and pragmatic for testing in this setting. It is also imperative that African investigators lead us in this important endeavor.


[Regenerative medicine for pancreatic beta cells].
Regenerative medicine involves varying degrees of interaction among many research domains. Regenerative medical therapies based on research into organogenesis and the regeneration of injured or dysfunctional tissue using cell therapy are being developed rapidly. For the treatment of diabetes mellitus (DM), pancreatic transplantation and islet (pancreatic endocrine) cell transplantation are considered to be one form of regenerative medicine to overcome pancreatic tissue dysfunction. Recently, the effective islet cell transplantation Edmonton protocol has been established, ushering in a new era in regenerative therapy for DM. However, unresolved problems remain, including a severe donor shortage and unexpected side effects with the longterm use of some immunosuppressive agents. With continuing advances and the clinical application of fundamental therapy for DM, a pancreatic islet cell transplantation or bioartificial pancreatic transplantation system, consisting of islet (pancreatic endocrine) cell purification, pancreatic cell proliferation techniques, immunoisolative membrane technology, and an appropriate transplantation procedure, will be effective. This paper focuses on applied research on human and/or porcine pancreatic cell purification, embryonic stem cell differentiation, and pancreatic stem cell differentiation into functional insulin-producing cells.


Bacteriology of acute otitis media: a new perspective.
Pathogenic bacteria were isolated from 90% of patients with acute otitis media. This higher-than-expected rate of positive cultures was probably related to the meticulous bacteriologic techniques used.


Predictive value of specific risk factors, symptoms and signs, in diagnosing obstructive sleep apnoea and its severity.
A positive diagnosis of obstructive sleep apnoea (OSA) is based on a combination of characteristic symptoms and polysomnographic findings. The present study evaluated the specificity and sensitivity of several risk factors, signs and symptoms in predicting an Apnoea Index in 86 patients referred to the sleep laboratory with suspected OSA. All 86 subjects completed a detailed questionnaire, were interviewed, underwent a brief physical examination, and then a whole-night polysomnographic study. Stepwise multiple regression analysis revealed that self reporting on apnoeas, neck circumference index (NCI), age, and a tendency to fall asleep unintentionally, were all significant positive predictors of apnoea index (AI), explaining 41.8% of the variability. The sensitivity of the model for predicting OSA (taking OSA as AI > 10) was 92.2%, specificity was 18.2% and the positive predictive value was 76.6%. Raising the cut-off AI values resulted in decreased sensitivity and increased specificity. Applying the predicting equation of AI to another group of 50 patients referred to the sleep laboratory with suspected OSA revealed similar results. However, running the equation on 105 offspring of OSA patients who did not complain of OSA-associated symptoms resulted in 32% sensitivity and 94% specificity in predicting OSA. It is concluded that questionnaires, interviews and physical examination, can only vaguely predict AI, and cannot replace polysomnographic recordings. However, the low rates of false negative in predicting AI > 10, and the low rates of false positive in predicting AI > 50, can be used for specific purposes.


Eukaryotic single-stranded DNA binding proteins: central factors in genome stability.
The single-stranded DNA binding proteins (SSBs) are required to maintain the integrity of the genome in all organisms. Replication protein A (RPA) is a nuclear SSB protein found in all eukaryotes and is required for multiple processes in DNA metabolism such as DNA replication, DNA repair, DNA recombination, telomere maintenance and DNA damage signalling. RPA is a heterotrimeric complex, binds ssDNA with high affinity, and interacts specifically with multiple proteins to fulfil its function in eukaryotes. RPA is phosphorylated in a cell cycle and DNA damage-dependent manner with evidence suggesting that phosphorylation has an important function in modulating the cellular DNA damage response. Considering the DNA-binding properties of RPA a mechanism of "molecular counting" to initiate DNA damage-dependent signalling is discussed. Recently a human homologue to the RPA2 subunit, called RPA4, was discovered and RPA4 can substitute for RPA2 in the RPA complex resulting in an "alternative" RPA (aRPA), which can bind to ssDNA with similar affinity as canonical RPA. Additional human SSBs, hSSB1 and hSSB2, were recently identified, with hSSB1 being localized in the nucleus and having implications in DNA repair. Mitochondrial SSBs (mtSSBs) have been found in all eukaryotes studied. mtSSBs are related to prokaryotic SSBs and essential to main the genome stability in eukaryotic mitochondria. Recently human mtSSB was identified as a novel binding partner of p53 and that it is able to stimulate the intrinsic exonuclease activity of p53. These findings and recent results associated with mutations in RPA suggest a link of SSBs to cancer.


The management of lower back pain using pilates method: assessment of content exercise reporting in RCTs.
To evaluate the quality of the reporting of exercise interventions with Pilates method for the treatment of lower back pain (LBP) in adults. Two independent evaluators selected randomized controlled trials (RCTs) of moderate and high methodological quality included in a Cochrane Systematic Review (SR) and from an additional updated search in the following databases: CENTRAL, MEDLINE, EMBASE, CINAHL, PEDro and SPORTDiscus. Three assessment tools (Consensus on Therapeutic Exercise Training (CONTENT) scale, Template for Intervention Description and Replication (TIDieR) checklist and Consensus on Exercise Reporting Template (CERT) checklist) were utilized by three pairs of two independent researchers trained. The scales' concordance was measured using the Kappa coefficient. Ten RCTs were included. The CONTENT scale score was 5.3 (± 1.33) out of 9 points; the TIDieR checklist was 8.5 (± 1.71) out of 12 points and the CERT checklist was 9.5 (± 3.62) out of 19 points. The CONTENT and CERT had moderate concordance, while there was fair concordance between the other tools. The overall reporting quality for the Pilates exercises in ten moderate-to-high quality RTCs for the management of LBP was low according to CONTENT scale and CERT checklist and high according to TIDieR checklist.Implications for RehabilitationReporting of Pilates exercise program in moderate-to-high quality RCTs for the management of lower back pain remains incomplete.Pilates exercise program should be personalized and contextualized to individual participants.There may be a need to consider adding to or combining the information available from various trials.


Noninvasive estimation of dynamic pressures in vitro and in vivo using the subharmonic response from microbubbles.
The purpose of this study was to develop and validate a noninvasive pressure estimation technique based on subharmonic emissions from a commercially available ultrasound contrast agent and scanner, unlike other studies that have either adopted a single-element transducer approach and/ or use of in-house contrast agents. Ambient pressures were varied in a closed-loop flow system between 0 and 120 mmHg and were recorded by a solid-state pressure catheter as the reference standard. Simultaneously, the ultrasound scanner was operated in pulse inversion mode transmitting at 2.5 MHz, and the unprocessed RF data were captured at different incident acoustic pressures (from 76 to 897 kPa). The subharmonic data for each pulse were extracted using band-pass filtering with averaging, and subsequently processed to eliminate noise. The incident acoustic pressure most sensitive to ambient pressure fluctuations was determined, and then the ambient pressure was tracked over 20 s. In vivo validation of this technique was performed in the left ventricle (LV) of 2 canines. In vitro, the subharmonic signal could track ambient pressure values with r(2) = 0.922 (p < 0.001), whereas in vivo, the subharmonic signal tracked the LV pressures with r(2) > 0.790 (p < 0.001) showing a maximum error of 2.84 mmHg compared with the reference standard. In conclusion, a subharmonic ultrasound-based pressure estimation technique, which can accurately track left ventricular pressures, has been established.


[Magnesium or proajmaline (neogilurytmal) in treatment of recurrences of paroxysmal supraventricular tachyarrhythmias?].
In order to compare the effectiveness of magnesium with that of proajmaline in preventing of recurrences of paroxysmal tachyarrhythmias. 60 consecutive patients were studied who had been previously treated for paroxysmal arterial fibrillation (PAF) or paroxysmal supraventricular tachycardia (PSVT). For the studies the patients were qualified who had been effectively treated during acute phase with intravenous magnesium (magnesium sulphate) and ajmaline (gilurytmal). The patients were randomly divided into three groups of 20 persons in each, every group received different oral treatment: in the first group--magnesium (magnesium carbonate) 3 x 0.6, in the second--proajmaline (neogilurytmal) in dose 3 x 20 mg, and in the third--neogilurytmal 2 x 20 mg and magnesium 2 x 0.6. During three months of observation the effectiveness of treatment and incidence of adverse effects were assessed. The following was found: neogilurytmal showed higher effectiveness (60%) than magnesium (30%), while the combined treatment demonstrated similar effectiveness (65%) as neogilurytmal alone with lower drug doses and lower incidence of bad tolerance manifestations (10% vs 20%).