Correlation study between bone metabolic markers, bone mineral density, and sarcopenia.

OBJECTIVE: To investigate the correlation between bone metabolism markers, bone mineral density (BMD), and sarcopenia. METHODS: A total of 331 consecutive patients aged ≥ 60 years who were hospitalized between November 2020 and December 2021 were enrolled. Participants were divided into sarcopenia and non-sarcopenia groups according to the Asian Working Group on Sarcopenia criteria (AWGS, 2019). The clinical data, bone metabolism markers (ß-CTX, N-MID, and TP1NP), and BMD were compared between the two groups. RESULTS: Age, ß-CTX, and N-MID of the sarcopenia group were higher than those of the non-sarcopenia group (P < 0.05), but the BMD T values were lower than those of the non-sarcopenia group (P < 0.05). Binary logistic regression analysis showed that increased femoral neck BMD (FNBMD) was a protective factor for sarcopenia, while increased ß-CTX was a risk factor. Pearson/Spearman correlation analysis showed that the diagnostic indices of sarcopenia were positively correlated with FNBMD and negatively correlated with ß-CTX and N-MID. Multiple linear regression analysis revealed that BMI and FNBMD significantly positively affected muscle strength and appendicular skeletal muscle mass (ASM). The FNBMD significantly positively affected physical performance, while ß-CTX significantly negatively affected muscle strength, ASM, and physical performance. CONCLUSION: Increased FNBMD may be a protective factor against sarcopenia, and increased ß-CTX may be a risk factor. The FNBMD significantly positively affected the diagnostic indices of sarcopenia, while ß-CTX significantly negatively affected them. BMD and bone metabolism marker levels may be considered in early screening for sarcopenia.

[The status and influencing factors of presenteeism among clinical nurses: a systematic review].

Objective: To systematically review the status and factors influencing presenteeism among clinical nurses. Methods: In December 2021, CNKI, CBM, Wanfang, VIP, Web of Science, PubMed, Embase, The Cochrane Library, CINAHL, PsyclNFO and other databases were electronically searched to cross sectional studies on the current situation and factors influencing the occurrence of presenteeism among clinical nurses. The search terms mainly included presenteeism, sick at work, Stanford Presenteeism Scale, nurse, level, risk factor, influence, et al. And the search time was from the establishment of the database to November 30, 2021. Literature screening, data extraction and evaluation of the risk of bias in the included literature were done independently by two researchers, and meta-analysis was performed using Stata 15.1 software. Results: A total of 29 studies involving 13 535 clinical nurses were included.The results of the meta-analysis showed that the score of presenteeism was 17.99 [95% CI (17.02-18.95), P =0.000]. Subgroup analysis showed that presenteeism scores were higher in articles published before 2020 (ES=19.28, 95%CI: 18.41-20.15, P=0.000) and in the group of nurses aged 36 to 40 years (ES=19.27, 95%CI: 17.35~21.19, P=0.000), female (ES= 17.04, 95%CI: 14.70-19.39, P=0.000), secondary school education (ES=21.01, 95%CI: 17.76-24.26, P= 0.007), married (ES=17.49, 95%CI: 15.13-19.85, P=0.000), working for 5 to 10 years (ES=17.78, 95%CI: 16.54-19.02, P=0.000), contract (ES=17.05, 95%CI: 15.23-18.87, P=0.000), working in pediatrics (ES= 16.65, 95% CI: 15.31-17.99, P=0.000) and European region (ES =21.21, 95% CI: 20.50-21.93, P=0.000) . Conclusion: Current evidence suggests that clinical nurses are at high risk of presenteeism, which is affected by variety of factors. The managers should pay attention to the physical and mental health of nurses, identify high-risk factors as early as possible and take measures to reduce the occurrence of presenteeism and improve the quality of nursing.

[The effectiveness of psychological intervention on nursing staff' compassion fatigue: a meta-analysis].

Objective: To systematically evaluate the impact of psychological intervention on nursing staff' compassion fatigue. Methods: From March to May 2020, PubMed, Cochrane Library, EMbase, Web of Science, CNKI, VIP, Wanfang and other databases were electronically searched to collect randomized controlled trials (RCTs) on the influence of psychological intervention on nursing staff' compassion fatigue with the main search terms including compassion fatigue, nurs*, psychological intervention, mental intervention, RCT and so on from inception to March 31, 2020. Screened literature, extracted data and assessed the risk of bias of included studies. The Stata 16.0 software was used to calculate the pooled effectiveness of psychological intervention on nursing staff' compassion fatigue. Results: All 13 RCTs were enrolled, including 940 nursing staff. Meta-analysis results demonstrated that the psychological intervention group was superior to the control group in the improvement of the compassion fatigue score (SMD=-0.96, 95%CI: -1.17-0.74, P=0.001) , compassion satisfaction score (SMD=0.61, 95%CI: 0.45-0.77, P=0.002) , burnout score (SMD=-0.46, 95%CI: -0.62-0.29, P=0.006) , secondary trauma score (SMD=-0.40, 95%CI: -0.68-0.12, P=0.020) , and the difference was statistically significant. Subgroup analysis found that the psychological intervention group was more effective than the control group in improving compassion satisfaction score, reducing burnout score and secondary trauma score, and the differences were statistically significant (P<0.05) in different intervention time (<8 weeks and ≥8 weeks) and intervention methods. Conclusion: The psychological intervention can improve the level of compassion satisfaction, and reduce the compassion fatigue among nursing staff, and have a certain preventive effect on compassion fatigue.

[Correlation between professional quality of life and social support of Chinese nurses: a meta-analysis].

Objective: To systematically evaluate the correlation between professional quality of life and social support of Chinese nurses based on Pearson and Spearman correlation coefficients. Methods: In databases including PubMed, Cochrane Library, CINAHL, Medline, CBM, CNKI、Wanfang, and other databases were searched by computer for the literatures on correlation between Chinese nurses' professional quality of life and social support from January 2005 to July 2020. The Chinese and English search terms are "nurse" "professional quality of life" "empathy satisfaction" "empathy fatigue" "professional quality of life" "ProQOL" "comparison satisfaction" "comparison fatigue" "social support" "competent social support" "SSRS" "PSSS", etc. Literatures were screened according to the inclusion and exclusion criteria. After evaluating quality and extracting data, meta-analysis was conducted using RevMan 5.3 software. Results: A total of 12 studies were included. The meta analysis showed that nurses' compassion satisfaction, burnout, secondary traumatic stress were related to social support, summary r were 0.35, -0.26 and -0.23 respectively. The correlation between compassion satisfaction and social support were increased with sample, the south was higher than the north, and comprehensive departments were higher than other departments (P<0.05) . The correlation between burnout and social support were increased with time and sample, and the south was higher than the north, oncology was higher than others, non-random sampling was higher than random sampling, using ProQOL and Perceived Social Support Scale (PSSS) was higher than Professional Quality of Life Scale (ProQOL) and Social Support Racting Scale (SSRS) (P<0.05) . The correlation coefficient between secondary traumatic stress and social support in oncology was higher than others, random sampling was higher than non-random sampling, using ProQOL and PSSS was higher than ProQOL and SSRS (P<0.05) . Conclusion: There is a positive and weak correlation between compassion satisfaction and social support, and a negative and weak correlation between burnout and secondary traumatic stress and social support. There are differences in different time, research design, region and department.

[Influencing factors for the use of earplugs in workers exposed to noise in a city].

OBJECTIVE: To investigate the current status of hearing loss and the use of earplugs in workers exposed to noise who have been provided earplugs in a city, as well as major influencing factors for the use of earplugs. METHODS: Cluster random sampling was used to conduct a questionnaire survey in workers exposed to noise who had been provided earplugs in 15 enterprises with noise exposure in a city from June to December, 2014. RESULTS: In the workers exposed to noise who had been provided earplugs, the rate of high-frequency anomaly in both ears was 57.8%, and the workers who kept wearing earplugs only accounted for 55.4%. The results of binary logistic regression analysis showed that the protective factors for the use of earplugs included workers' own feeling of hearing condition (OR=1.704), comfort of earplugs (OR= 1.892), enterprise's inspection of the use of earplugs (OR=1.461), workers' knowledge of the function and usage of earplugs (OR=1.581), workers' understanding of the necessity of earplugs (OR=4.482), workers' initiative to search for related data (OR=4.029), the use of earplugs by colleagues (OR=5.071), and reminders from family members or friends (OR=2.678) (all P<0.05). CONCLUSION: The workers exposed to noise in this city have a high rate of abnormal hearing, and only half of the workers keep wearing earplugs during work. The use of earplugs is related to the factors including workers' own feeling of hearing condition, comfort of earplugs, workers' knowledge of protection, the enterprise' s management of hearing protection, and environmental support.

Cyclosporine A and tacrolimus combined with enteric-coated mycophenolate sodium influence the plasma mycophenolic acid concentration - a randomised controlled trial in Chinese live related donor kidney transplant recipients.

OBJECTIVE: To observe the different concentrations of mycophenolic acid (MPA) during the early post-transplant phase for various combinations of cyclosporine A (CsA) and tacrolimus (Tac) with enteric-coated mycophenolate sodium (EC-MPS). METHOD: A total of 42 Chinese adults receiving live related donor kidney transplants were studied. All received a triple immunosuppressive regimen of EC-MPS, CsA/Tac and corticosteroids and were divided randomly into CsA (n = 21) and Tac (n = 21) combination groups. The dosage of EC-MPS was the same (1440 mg/day) in the two groups. The MPA concentration was evaluated with an enzyme-multiplied immunoassay technique (EMIT) and the pharmacokinetic characteristics were investigated in both groups. RESULTS: The mean maximum plasma concentrations (Cmax ) of MPA in the CsA and Tac groups were 11.365 ± 9.522 and 9.748 ± 7.523 µg/ml, respectively (p = 0.137). The maximum times to Cmax (Tmax ) were 2.54 ± 1.53 and 2.67 ± 1.08 h, respectively (p = 0.341). The mean MPA 12-h areas under the curve (MPA-AUC0-12 h ) were 59.463 ± 16.252 and 77.535 ± 33.615 µg h/ml (p = 0.003) and the mean residence times (MRT) were 3.71 ± 0.829 and 3.928 ± 0.923 h (p = 0.038). CONCLUSION: Combined with the same EC-MPS dosage (1440 mg/day), the MPA-AUC0-12 of the Tac group was higher than that of the CsA group, and the Tac group had a longer MRT after kidney transplantation. Our data indicate that the concentration of MPA should be monitored in clinical therapy when EC-MPS is combined with different calcineurin inhibitors to reduce acute allograft rejection and avoid adverse events.

[Establishment of leukemia cell model with inducible AML1-ETO expression and its effect on fatty acid metabolism in leukemia cells].

Objective: To investigate the effect of the AML1-ETO (AE) fusion gene on the biological function of U937 leukemia cells by establishing a leukemia cell model that induces AE fusion gene expression. Methods: The doxycycline (Dox) -dependent expression of the AE fusion gene in the U937 cell line (U937-AE) were established using a lentivirus vector system. The Cell Counting Kit 8 methods, including the PI and sidanilide induction, were used to detect cell proliferation, cell cycle-induced differentiation assays, respectively. The effect of the AE fusion gene on the biological function of U937-AE cells was preliminarily explored using transcriptome sequencing and metabonomic sequencing. Results: ①The Dox-dependent Tet-on regulatory system was successfully constructed to regulate the stable AE fusion gene expression in U937-AE cells. ②Cell proliferation slowed down and the cell proliferation rate with AE expression (3.47±0.07) was lower than AE non-expression (3.86 ± 0.05) after inducing the AE fusion gene expression for 24 h (P<0.05). The proportion of cells in the G(0)/G(1) phase in the cell cycle increased, with AE expression [ (63.45±3.10) %) ] was higher than AE non-expression [ (41.36± 9.56) %] (P<0.05). The proportion of cells expressing CD13 and CD14 decreased with the expression of AE. The AE negative group is significantly higher than the AE positive group (P<0.05). ③The enrichment analysis of the transcriptome sequencing gene set revealed significantly enriched quiescence, nuclear factor kappa-light-chain-enhancer of activated B cells, interferon-α/γ, and other inflammatory response and immune regulation signals after AE expression. ④Disorder of fatty acid metabolism of U937-AE cells occurred under the influence of AE. The concentration of the medium and short-chain fatty acid acylcarnitine metabolites decreased in cells with AE expressing, propionyl L-carnitine, wherein those with AE expression (0.46±0.13) were lower than those with AE non-expression (1.00±0.27) (P<0.05). The metabolite concentration of some long-chain fatty acid acylcarnitine increased in cells with AE expressing tetradecanoyl carnitine, wherein those with AE expression (1.26±0.01) were higher than those with AE non-expression (1.00±0.05) (P<0.05) . Conclusion: This study successfully established a leukemia cell model that can induce AE expression. The AE expression blocked the cell cycle and inhibited cell differentiation. The gene sets related to the inflammatory reactions was significantly enriched in U937-AE cells that express AE, and fatty acid metabolism was disordered.

Realization of effective light trapping and omnidirectional antireflection in smooth surface silicon nanowire arrays.

We have successfully fabricated well-ordered silicon nanowire (SiNW) arrays of smooth surface by using a low-cost and facile Ag-assisted chemical etching technique. We have experimentally found that the reflectance can be significantly suppressed (<1%) over a wide solar spectrum (300-1000 nm) in the as-grown samples. Also, based on our bundled model, we have used rigorous coupled-wave analysis to simulate the reflectance in SiNW arrays, and found that the calculated results are in good agreement with the experimental data. From a further simulation study on the light absorption in SiNW arrays, we have obtained a photocurrent enhancement of up to 425% per unit volume of material as compared to crystalline Si, implying that effective light trapping can be realized in the as-grown samples. In addition, we have demonstrated experimentally and theoretically that the as-grown samples have an omnidirectional high-efficiency antireflection property.

[Short-term outcomes of single incision plus one port laparoscopic surgery for colorectal cancer].

Objective: Although single port laparoscopic surgery has achieved good clinical results, many surgeons are discouraged by the difficulties of operation, conflict of instruments, lack of antagonistic traction, and straight-line perspective. Therefore, some surgeons have proposed a single incision plus one hole laparoscopic surgery (SILS+1) surgical method. This study explored the safety and feasibility of SILS+1 for radical resection of colorectal cancer. Methods: A descriptive cohort study was carried out. The clinical data, including the operation, pathology and recovery situation, of 178 patients with colorectal cancer undergoing SILS+1 at Department of General Surgery, Nanfang Hospital, Southern Medical University from March 2018 to January 2019 were prospectively collected and retrospectively analyzed. Clavien-Dindo criteria was used for postoperative complication evaluation and visual analog scale was used for pain standard. Follow-up studies were conducted through outpatient service or telephone and the follow-up period was up to May 2019. Results: A total of 178 patients with colorectal cancer underwent SILS+1, including 111 male patients (62.4%) with an average age of 59 years. Eleven (6.2%) patients received added 1-3 operation ports during operation, and 1 patient was converted to open surgery due to ileocolic artery hemorrhage. The operative time was (135.2±42.3) minutes. The intraoperative blood loss was (34.6±35.5) ml. The number of harvested lymph nodes was 33.1±17.6. The distal margin was (4.7±17.8) cm. The proximal margin was (10.2±5.3) cm. Operation-related complications were observed in 16 patients (9.0%) within 30 days after the operation, of whom 6 had Clavien-Dindo III complications (3.4%). The postoperative pain scores were lower than 3. The average postoperative hospital stay was (5.6±2.6) days. Three patients (1.7%) returned to hospital within 30 days after operation due to intestinal obstruction and infection around stoma. The cosmetic evaluation of all the patients was basically satisfied. Conclusion: SILS+1 is safe and feasible in the treatment of colorectal cancer, and can reduce the postoperative pain.

Domains of the rat rDNA promoter must be aligned stereospecifically.

Efficient transcription from the rat rDNA promoter results from an undefined interaction between the core (CPE) and upstream (UPE) promoter elements or the protein complexes which form on them. These interactions were demonstrated by the behavior of promoters that contained either linker-scanning or deletion mutations of the UPE in combination with point mutations of the CPE (bidomain mutants). In vivo transcription experiments using point mutations within the CPE (G----A mutation at either -16 or -7) demonstrated that the CPE may in fact consist of two domains. Whereas both of these mutants were rescued by the addition of UBF to in vitro transcription reactions, the CPE mutant -7A/G was inactive in vivo. Experiments with these bidomain mutants demonstrated that the UPE was required for the rescue of the CPE mutants. We also examined the hypothesis that this interaction might require a stereospecific alignment of the promoter elements. Our results indicate that the promoter consists of several domains with differing responses to mutations that alter the distance between, or within, the promoter elements. For example, the insertion or deletion of half-multiples of the helical repeat distance between -167 and -147 had no significant effect on transcription. On the other hand, some sites were sensitive to deletions of any size but not to insertions of up to 20 bp. The analyses of two sites yielded results suggesting that they lay between domains of the promoter that must be on the same side of the DNA helix for promoter activity. The first of these sites mapped between -106 and -95.(ABSTRACT TRUNCATED AT 250 WORDS)

[The relationship between type ⠡ diabetes mellitus and hearing loss：a Meta-analysi].

Objective:Analysis of the relationship between type 2 diabetes and hearing loss through the available literature.Method:According to the main keyword retrieval databases at home and abroad,and in accordance with the inclusion and exclusion criteria for the literature,the listening incidence rate,OR value,the frequency of pure tone hearing threshold,tests the average standard deviation were chose as the evaluation index. Meta analysis was carried out using Rev Man5.3 software. Result:According to the inclusion and exclusion criteria,the articles were screened and included in the final to meet the requirements of the 17 studies. The prevalence of hearing loss in patients with type II diabetes mellitus was 21.0%-74.6%,which was 2.54 times higher than the control group. The difference was statistically significant between the two groups(OR=2.54,95% CI 1.85-3.49,P<0.000 01). In addition,the hearing threshold of all frequencies in type Ⅱ diabetes group was higher than that of control group(Z=5.03,P<0.000 01). Conclusion:Type 2 diabetes can affect hearing loss,especially at the high frequencies.

[Screening on key techniques used for surveillance and disposal of public health emergencies].

Objective: To explore the key techniques used for surveillance and disposal of infectious diseases, food poisoning and hospital infection to improve the ability of surveillance and disposal on public health emergency. Methods: Framework on surveillance and disposal of infectious diseases, food poisoning and hospital infection was set up, based on literature review and expert group discussion. Delphi method and technique for order preference by similarity to ideal solution comprehensive evaluation method were used for ordering preference by similarity, to screen key techniques set for surveillance and disposal of the above said events. Results: Framework to be used for selecting key techniques was designed, based on the classification of emergency events, processing cycle of emergency events and level of techniques. Twenty six public health experts were selected for a 2-round consultation, with their authority as 0.796. Ten key techniques with important significance for surveillance and disposal of infectious diseases, food poisoning and hospital infection were selected from each event. Among these key techniques, the early-warning system was recognized as the key technique, important for the surveillance and disposal of all three emergency events. Items as technology used for unknown pathogenic microorganism detection, personal protection, gene sequencing and tracing technology, microorganism molecular typing technology, nucleic acid detection technology etc. were the key techniques and need to develop for the surveillance and disposal of infectious diseases and iatrogenic infection. Data regarding key technologies on security and privacy, early warning and forecasting, field rapid detection were sorted out that all in need to improve the surveillance programs on disposal of infectious diseases and food poisoning. Data exchange appeared another key technique on infectious diseases, with toxin detection and other 5 techniques the key techniques for food poisoning. Data collection, standardization and other 3 techniques appeared as the key techniques on iatrogenic infection. Conclusions: Key techniques were selected in this study to meet the requirements of surveillance, disposal of infectious disease, food poisoning and iatrogenic infection. Authority of the experts got improved in the two-round consultation, ensuring the reliability of the results of screening and providing a scientific basis for the further research and priority development of the key techniques.

Rapid, small-scale RNA isolation from tissue culture cells.

A rapid and simple protocol for the isolation of RNA from transfected tissue culture cells is described. The protocol employs a guanidinium thiocyanate/phenol mixture to lyse cells directly from tissue culture plates and extract the total RNA. A total of six simple steps, which can be accomplished within 2.5 hours, are required. The protocol reproducibly yields 20-40 micrograms RNA from 0.5 x 10(6) - 1 x 10(6) cells per sample. The quality of the RNA obtained is sufficient for reverse transcriptase assays such as oligonucleotide-directed primer extension and random-primed cDNA synthesis.

Gene cluster rpoBC1C2 in cyanobacteria does not constitute an operon.

The core enzyme of the cyanobacterial DNA-dependent RNA polymerase contains a unique component, gamma, which is absent from the corresponding enzymes of other eubacteria. In the heterocystous cyanobacterium Nostoc commune the gene encoding gamma, rpoC1, is immediately adjacent to, and downstream of, rpoB. The rpoC1 gene, and a 3' adjacent gene, rpoC2, correspond to the single rpoC gene found in Escherichia coli with respect to those domains conserved within their translational products. Northern analysis and primer extension assay show that in N. commune, rpoC1 and rpoC2 are transcribed separately from rpoB. The promoter of rpoC1C2 can direct the expression of a promotorless lacZ gene in E. coli. As a consequence, cyanobacterial rpo gene expression is distinct from the mode of cotranscription described for the equivalent sequences found in other eubacteria, archaebacteria, and plant chloroplasts. Also in this paper, a simple protocol for RNA isolation, which should be applicable for RNA isolation from plant cells, is presented.

Quick screening of plasmid deletion clones carrying inserts of desired sizes for DNA sequencing.

Plasmids pWQX001 and pWQX005, constructed from pGEM-4 with an insert of 6.5 kb, were unidirectionally digested with exonuclease III and exonuclease VII. The DNA digests were ligated and used to transform competent cells of Escherichia coli DH5-alpha. The size of the deletion plasmid carried by each transformant was estimated through agarose gel electrophoresis of crude lysates without any purification of the plasmid DNA. Colonies carrying plasmid DNAs with different deletions of the insert were grown and their DNAs were purified through a miniprocedure. The size of each purified plasmid DNA was determined accurately after linearization of the plasmid with an appropriate restriction endonuclease. The remainder of the DNA preparation was sufficiently pure to be sequenced using Sanger's dideoxynucleotide chain termination method. An easy, quick procedure is described for the preliminary selection of templates for DNA sequencing after construction of deletion clones of recombinant plasmid DNA using exonuclease III and exonuclease VII. This procedure permits a rapid screening of large numbers of colonies and selection of those carrying plasmid DNAs with inserts of the desired sizes for sequencing. This procedure does not require purification of the deletion plasmid DNA.

Cyanobacterial RNA polymerase genes rpoC1 and rpoC2 correspond to rpoC of Escherichia coli.

The DNA-dependent RNA polymerase (ribonucleoside triphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) of cyanobacteria contains a unique core component, gamma, which is absent from the RNA polymerases of other eubacteria (G. J. Schneider, N. E. Tumer, C. Richaud, G. Borbely, and R. Haselkorn, J. Biol. Chem. 262:14633-14639, 1987). We present the complete nucleotide sequence of rpoC1, the gene encoding the gamma subunit, from the heterocystous cyanobacterium Nostoc commune UTEX 584. The derived amino acid sequence of gamma (621 residues) corresponds with the amino-terminal portion of the beta' polypeptide of Escherichia coli RNA polymerase. A second gene in N. commune UTEX 584, rpoC2, encodes a protein which shows correspondence with the carboxy-terminal portion of the E. coli beta' subunit. The rpoBC1C2 genes of N. commune UTEX 584 are present in single copies and are arranged in the order rpoBC1C2, and the coding regions are separated by short AT-rich spacer regions which have the potential to form very stable secondary structures. Our data indicate the occurrence of divergent evolution of structure in the eubacterial DNA-dependent RNA polymerase.

Complementary in vivo and in vitro analyses of the interactions between the cis-acting elements of the rat rDNA promoter.

Two transcription factors, rat UBF (rUBF) and rat SL-1 are required for the efficient transcription of the rat promoter in vitro. In vitro studies have established that two broadly defined cis-acting domains, the core promoter element and the upstream promoter element, cooperate to direct correct transcription by RNA polymerase I. The ability of UBF to bind to two linker-scanning mutants of the upstream promoter element, which did not respond to the addition of UBF in in vitro transcription assays, was assessed by DNase footprinting. UBF protected the same region of the promoter in the linker-scanning mutant in BSM 129/124 as it did in the wild-type, but did not yield a typical footprint over the promoter in the linker-scanning mutant BSM 106/101. Previously we reported that promoters with mutant core promoters elements, either the guanine at -16 or -7 substituted by an adenine, were inactive in vitro unless the assays were supplemented with UBF. Those results suggested that the binding of UBF upstream of the core was required for the promotion of transcription. The interactions between the core and upstream promoter elements were assessed by constructing double mutants of the promoter. In two constructs the conserved guanines at either -16 or -7 were altered in a deletion mutant (-86) that did not respond to UBF. In a third construct the guanine at -16 in BSM 129/124 was changed to an adenine. These bidomain mutant constructs did not respond to the addition of UBF in an in vitro transcription assay, confirming that the rescue of the core promoter mutants requires an intact and functional upstream promoter element.(ABSTRACT TRUNCATED AT 250 WORDS)