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A proof-of-concept study is reported for the differentiation between microcolonies of Enterobacter sakazakii and Klebsiella pneumoniae by means of a novel sample preparation for infrared (IR) analysis. A disposable, IR-transparent, microporous (0.2-µm pores), hydrophobic, polyethylene (PE) membrane (51 µm thick) was plasma treated under an oxygen atmosphere and used to (i) filter (or print microarrays of) dilute aqueous foodborne bacterial suspensions and (ii) subsequently grow bacterial microcolonies when the treated, hydrophilic PE membrane was placed over brain heart infusion agar medium and incubated. Because this unique membrane is transparent to IR light, isolated microcolonies (200 µm) of bacterial cells grown on this PE substrate for the first time could be directly fingerprinted by IR microspectroscopy in the transmission mode. Hence, time-consuming bacterial cell transfer from culture plates to an IR sample holder for subsequent measurement by IR spectroscopy was eliminated. Multivariate analysis of the observed IR spectra for microcolonies allowed the rapid differentiation between E. sakazakii and K. pneumoniae. [ABSTRACT FROM AUTHOR]

This article presents news from the U.S. Centers for Disease Control and Prevention (CDC). This study looked at Salmonella infection outbreaks in the United States due to raw tomato consumption at restaurants in 2005-2006. There were four large outbreaks in the time period that resulted in 459 confirmed cases in 21 states. Investigation by the CDC determined that the tomatoes had been shipped from fields in Florida, Ohio and Virginia and says that these outbreaks show the need to research policies that will prevent Salmonella contamination early in the production and packing process.

The article describes the epidemiologic, environmental and laboratory investigations of the four large multistate outbreaks of Salmonella infections associated with eating raw tomatoes at restaurants in the U.S. from 2005-2006. During 2005-2006, there were 459 culture-confirmed cases of salmonellosis in 21 states. The investigations conducted by state and local health departments, national food safety agencies and CDC determined that the tomatoes had been supplied to restaurants either whole or precut from tomato fields in Florida, Ohio and Virginia.

Background: Postpartum depression (PPD) is a common mental health condition that affects women in a silent and covert way and is not clearly visible to the community or to health care providers. Untreated PPD has significant and long-term consequences on the mother and their child. This study aims to assess the risk of postpartum depression among women in the Emirate of Abu Dhabi and its determinants., Method: This is a questionnaire-based cross-sectional study conducted at primary healthcare centers in the Emirate of Abu Dhabi. The target population is women visiting the well-child vaccination clinics for their infants' vaccination. The questionnaire used consisted of socio-demographic characteristics, important histories such as obstetric, medical, and social histories, and the Edinburgh Post Partum depression scale EPPS. EPPS is a validated tool used to evaluate the probability of postpartum depression., Results: The probability of postpartum depression for women visiting the well child care clinics in the Emirate of Abu Dhabi during the study period was 35%, One-third. 10% had high risk, 7% had moderate risk, and 18% had mild risk. Using logistic and linear regression, there was an association identified between postpartum depression risk and the presence of weight concern and employment status OR 5.499(2.618-11.548) and OR 0.483 (0.246-0.951), respectively (P < 0.005). From the total sample, 3.7% responded quite often or sometimes to the question of having the intention to harm themselves., Conclusion: EPDS is recommended to be used routinely to screen women in the postnatal period. This high prevalence of risk of postpartum depression in the UAE (One in three women) calls for a well-prepared healthcare system and community. Healthcare providers need to be prepared with better knowledge, practice, and management strategies to care for these women, for early identification and management. Further studies should be undertaken to achieve effective strategies to reduce the incidence of this condition., (© 2024. The Author(s).)

Omics technologies provide useful tools for the identification of novel biomarkers in many diseases, including breast cancer, which is the most diagnosed cancer in women worldwide. We and others have reported a central role for the actin-bundling protein (fascin) in regulating breast cancer disease progression at different levels. However, whether fascin expression promotes metabolic molecules that could predict disease progression has not been fully elucidated. Here, fascin expression was manipulated via knockdown (fascin KD+NORF ) and rescue (fascin KD+FORF ) in the naturally fascin-positive (fascin pos+NORF ) MDA-MB-231 breast cancer cells. Whether fascin dysregulates metabolic profiles that are associated with disease progression was assessed using untargeted metabolomics analyses via liquid chromatography-mass spectrometry. Overall, 12,226 metabolic features were detected in the tested cell pellets. Fascin pos+NORF cell pellets showed 2510 and 3804 significantly dysregulated metabolites compared to their fascin KD+NORF counterparts. Fascin rescue (fascin KD+FORF ) revealed 2710 significantly dysregulated cellular metabolites compared to fascin KD+NORF counterparts. A total of 101 overlapped cellular metabolites between fascin KD+FORF and fascin pos+NORF were significantly dysregulated in the fascin KD+NORF cells. Analysis of the significantly dysregulated metabolites by fascin expression revealed their involvement in the metabolism of sphingolipid, phenylalanine, tyrosine, and tryptophan biosynthesis, and pantothenate and CoA biosynthesis, which are critical pathways for breast cancer progression. Our findings of fascin-mediated alteration of metabolic pathways could be used as putative poor prognostic biomarkers and highlight other underlying mechanisms of fascin contribution to breast cancer progression.

Hypertension is one of the most common and complicated disorders associated with genetic and environmental risk factors. The angiotensin-converting enzyme (ACE) is important in the renin-angiotensin-system pathway. The gene expression of ACE has been investigated as a possible hypertension marker. This study investigates the association between polymorphisms within the ACE1 and ACE2 genes and hypertension susceptibility in a Jordanian population. The study comprised a total of 200 hypertensive patients and 180 healthy controls. A polymerase chain reaction (PCR) was performed to genotype the candidate polymorphism (rs4646994) of the ACE1gene. The Luminex DNA array technique was used for genotyping SNPs (rs4359, rs4344, rs4341, rs4343, and rs2106809) of the ACE1 and ACE2 genes. Our findings suggest no association between SNPs and hypertension regarding allelic and genotypic frequencies. However, rs4359 was significantly associated with diet (pP = 0.049), know HTN (P = 0.042), and number of years DM (P = 0.003). rs4341 was associated with diet (P = 0.032), peripheral vascular disease (P = 0.005), and chronic kidney disease (p = 0.049). While rs4343 was associated with diet (P = 0.031), diabetes mellitus (P = 0.032), and other medication (P = 0.025). Furthermore, the haplotypes of four SNPs of the ACE1 gene showed no significant association with HTN patients and healthy controls. Our findings indicate no association between the polymorphisms in the ACE gene and the risk of hypertension development in the Jordanian adult population., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 AL-Eitan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Fascin expression has commonly been observed in certain subtypes of breast cancer, where its expression is associated with poor clinical outcome. However, its role in normal mammary gland development has not been elucidated. Here, we used a fascin knockout mouse model to assess its role in normal mammary gland morphogenesis and lactation. Fascin knockout was not embryonically lethal, and its effect on the litter size or condition at birth was minimal. However, litter survival until the weaning stage significantly depended on fascin expression solely in the nursing dams. Accordingly, pups that nursed from fascin -/- dams had smaller milk spots in their abdomen, suggesting a lactation defect in the nursing dams. Mammary gland whole-mounts of pregnant and lactating fascin -/- mice showed significantly reduced side branching and alveologenesis. Despite a typical composition of basal, luminal, and stromal subsets of mammary cells and normal ductal architecture of myoepithelial and luminal layers, the percentage of alveolar progenitors (ALDH + ) in fascin -/- epithelial fraction was significantly reduced. Further in-depth analyses of fascin -/- mammary glands showed a significant reduction in the expression of Elf5, the master regulator of alveologenesis, and a decrease in the activity of its downstream target p-STAT5. In agreement, there was a significant reduction in the expression of the milk proteins, whey acidic protein (WAP), and β-casein in fascin -/- mammary glands. Collectively, our data demonstrate, for the first time, the physiological role of fascin in normal mammary gland lactogenesis, an addition that could reveal its contribution to breast cancer initiation and progression., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Recent years have witnessed major progress in development of novel therapeutic agents such as chemotherapy, targeted therapy and immune checkpoint inhibitors for breast cancer. However, cancer-related death remains high especially in triple-negative breast cancer (TNBC) due limited therapeutic options. Development of targeted therapies for TNBC requires better understanding of biology and signaling networks that promote disease progression. Fascin, an actin bundling protein, was identified as a key regulator of many signaling pathways that contribute to breast cancer progression. Herein, fascin ShRNA was used to generate stable fascin knockdown (FSCN1 KD ) in the MDA-MB-231 TNBC cell line and then were subjected to comprehensive mRNA and miRNA transcriptome analysis. We identified 129 upregulated and 114 downregulated mRNA transcripts, while 14 miRNAs were differentially expressed in FSCN1 KD . Ingenuity pathway analysis (IPA) was used to predict the impact of differentially expressed transcripts on signaling pathways and functional categories and to construct miRNA-mRNA regulatory networks in the context of FSCN1 knockdown. Compared to FSCN1 KD , fascin-positive (FSCN1 CON ) breast cancer cells showed enrichment in genes promoting cellular proliferation, migration, survival, DNA replication and repair. Expression of FSCN1 high (identified in BRCA dataset from TCGA) in conjunction with elevated expression of the top 10 upregulated or decreased expression of the top 10 downregulated genes (identified in our FSCN1 CON vs. FSCN1 KD ) correlates with worst survival outcome. Taken together, these data confirmed fascin's role in promoting TNBC progression, and identified a novel opportunity for therapeutic interventions via targeting those FSCN1-related transcripts.

Cancer stem cells (CSCs), a rare population of tumor cells with high self-renewability potential, have gained increasing attention due to their contribution to chemoresistance and metastasis. We have previously demonstrated a critical role for the actin-bundling protein (fascin) in mediating breast cancer chemoresistance through activation of focal adhesion kinase (FAK). The latter is known to trigger the β-catenin signaling pathway. Whether fascin activation of FAK would ultimately trigger β-catenin signaling pathway has not been elucidated. Here, we assessed the effect of fascin manipulation in breast cancer cells on triggering β-catenin downstream targets and its dependence on FAK. Gain and loss of fascin expression showed its direct effect on the constitutive expression of β-catenin downstream targets and enhancement of self-renewability. In addition, fascin was essential for glycogen synthase kinase 3β inhibitor-mediated inducible expression and function of the β-catenin downstream targets. Importantly, fascin-mediated constitutive and inducible expression of β-catenin downstream targets, as well as its subsequent effect on CSC function, was at least partially FAK dependent. To assess the clinical relevance of the in vitro findings, we evaluated the consequence of fascin, FAK, and β-catenin downstream target coexpression on the outcome of breast cancer patient survival. Patients with coexpression of fascin high and FAK high or high β-catenin downstream targets showed the worst survival outcome, whereas in fascin low , patient coexpression of FAK high or high β-catenin targets had less significant effect on the survival. Altogether, our data demonstrated the critical role of fascin-mediated β-catenin activation and its dependence on intact FAK signaling to promote breast CSC function. These findings suggest that targeting of fascin-FAK-β-catenin axis may provide a novel therapeutic approach for eradication of breast cancer from the root., (Copyright © 2020 Barnawi, Al-Khaldi, Bakheet, Fallatah, Alaiya, Ghebeh and Al-Alwan.)

Breast cancer remains the second cause of tumor-related mortality in women worldwide mainly due to chemoresistance and metastasis. The chemoresistance and metastasis are attributed to a rare subpopulation with enriched stem-like characteristics, thus called Cancer Stem Cells (CSCs). We have previously reported aberrant expression of the actin-bundling protein (fascin) in breast cancer cells, which enhances their chemoresistance, metastasis and enriches CSC population. The intracellular mechanisms that link fascin with its downstream effectors are not fully elucidated. Here, loss and gain of function approaches in two different breast cancer models were used to understand how fascin promotes disease progression. Importantly, findings were aligned with expression data from actual breast cancer patients. Expression profiling of a large breast cancer dataset (TCGA, 530 patients) showed statistically significant correlation between fascin expression and a key adherence molecule, β1 integrin (ITGB1). In vitro manipulation of fascin expression in breast cancer cells exhibited its direct effect on ITGB1 expression. Fascin-mediated regulation of ITGB1 was critical for several breast cancer cell functions including adhesion to different extracellular matrix, self-renewability and chemoresistance. Importantly, there was a significant relationship between fascin and ITGB1 co-expression and short disease-free as well as overall survival in chemo-treated breast cancer patients. This novel role of fascin effect on ITGB1 expression and its outcome on cell self-renewability and chemoresistance strongly encourages for dual targeting of fascin-ITGB1 axis as a therapeutic approach to halt breast cancer progression and eradicate it from the root., (© 2019 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.)

Objectives: To determine the characteristics of patients presenting with chronic pain in a primary health care setting in Oman., Methods: A retrospective cross-sectional study was carried out including all patients aged ≥ 18 years who attended Sultan Qaboos University Health Center during 2010. Patients were identified to have chronic pain if they were prescribed an analgesic medication for at least three months. Patients were compared to a control group which consisted of age- and gender-matched patients with no chronic pain., Results: Out of 6 609 patients, 241 (3.6%) were found to have chronic pain. The mean age of patients with chronic pain was 54.0±13.0 years. The majority of patients were female (n = 174; 72.1%), and most were Omani (n = 201; 83.4%). The prevalence of chronic pain was found to be significantly higher among females compared to males (4.5% vs. 2.5%; p < 0.001) and also among Omani nationals to non-nationals (83.4% vs. 70.1%; p < 0.001). Chronic pain was significantly associated with the following comorbidities; diabetes (33.1% vs. 20.7%; p < 0.001), obesity (35.2% vs. 26.5%; p = 0.001), and hypertension (51.0% vs. 38.5%; p = 0.002). Osteoarthritis was the most common pain condition (n = 104; 43.1%). Diclofenac was the most commonly prescribed drug (n = 168; 69.7%)., Conclusions: The findings of our study point towards a higher prevalence of chronic pain in Omani females. These patients were also found to have a higher prevalence of other common comorbid conditions.

An emerging dogma shows that tumors are initiated and maintained by a subpopulation of cancer cells that hijack some stem cell features and thus referred to as "cancer stem cells" (CSCs). The exact mechanism that regulates the maintenance of CSC pool remains largely unknown. Fascin is an actin-bundling protein that we have previously demonstrated to be a major regulator of breast cancer chemoresistance and metastasis, two cardinal features of CSCs. Here, we manipulated fascin expression in breast cancer cell lines and used several in vitro and in vivo approaches to examine the relationship between fascin expression and breast CSCs. Fascin knockdown significantly reduced stem cell-like phenotype (CD44 hi /CD24 lo and ALDH + ) and reversal of epithelial to mesenchymal transition. Interestingly, expression of the embryonic stem cell transcriptional factors (Oct4, Nanog, Sox2, and Klf4) was significantly reduced when fascin expression was down-regulated. Functionally, fascin-knockdown cells were less competent in forming colonies and tumorspheres, consistent with lower basal self-renewal activity and higher susceptibility to chemotherapy. Fascin effect on CSC chemoresistance and self-renewability was associated with Notch signaling. Activation of Notch induced the relevant downstream targets predominantly in the fascin-positive cells. Limiting-dilution xenotransplantation assay showed higher frequency of tumor-initiating cells in the fascin-positive group. Collectively, our data demonstrated fascin as a critical regulator of breast CSC pool at least partially via activation of the Notch self-renewal signaling pathway and modification of the expression embryonic transcriptional factors. Targeting fascin may halt CSCs and thus presents a novel therapeutic approach for effective treatment of breast cancer. Stem Cells 2016;34:2799-2813 Video Highlight: https://youtu.be/GxS4fJ&#95;Ow-o., (© 2016 AlphaMed Press.)

A novel application of mid-infrared chemical imaging (IRCI) for the fluorophore-free detection and identification of mycoplasma species is reported for the first time. The PCR-amplified biotinylated targets hybridized to microarray probes were treated with streptavidin-gold nanoparticles followed by silver enhancement. This modification has the potential to expand the implementation of DNA microarray techniques in laboratories involved in the detection of cell substrates, other biological products, and clinical materials for the presence of mycoplasmas., (Published by Elsevier B.V.)

Francisella tularensis is a Gram-negative bacterium that can cause gastrointestinal or oropharyngeal tularemia in humans from ingestion of contaminated food or water. Despite the potential for accidental or intentional contamination of foods with F. tularensis, there are few studies on the long-term survivability of this organism in food matrices. Infant formula has previously been implicated as a vehicle for the transmission of a variety of bacterial pathogens in infants. In this study, we investigated the survival of F. tularensis in dehydrated infant formula under various storage conditions. F. tularensis was stored for up to 12 weeks in dehydrated infant formula in an ambient air, dry or nitrogen atmosphere. Viable counts of fresh F. tularensis at 12 weeks in infant formula revealed a 4.15, 3.37 and 3.72-log decrease in ambient air, dry and nitrogen atmosphere, respectively. D-values were calculated (in weeks) as 3.99, 4.68 and 4.47 in air, dry and nitrogen atmosphere, respectively.

Objectives: To evaluate the prevalence of hypertension, its control and management at Sultan Qaboos University (SQU) Health Centre, Oman., Methods: This was a retrospective cross-sectional study, in which were enrolled all the subjects (≥18 years), with the diagnosis of essential hypertension, who attended the SQU Health Centre between 1998 and 2002. The systolic and diastolic blood pressure (BP) values of the last three visits were used for analysis. BP control was defined using the Joint National Committee (JNC-7) criteria, <140 mmHg and <90 mmHg for systolic and diastolic BPs, respectively. Analyses were performed using univariate statistics., Results: Among the 7,702 medical records reviewed, the prevalence of hypertension was 2.4% (n = 187). The overall mean age of the cohort was 55±11 years, 54% (n = 101) were females, and majority of the subjects were Omanis (n = 123; 66%). The proportion of subjects who had their BP controlled was 41% (n = 77) with Omanis significantly less likely to have their BP controlled compared to non-Omanis (53% versus 35%; p = 0.017). The majority of the subjects were on mono (n = 131; 70%) followed by dual (n = 50; 27%) anti-hypertensive therapies. The most frequent mono anti-hypertensive therapies were B-blockers (n = 64; 34%) and angiotensin-converting enzyme (ACE) inhibitors (n = 47; 25%). Among the dual combination therapies, the most common prescribed regimens were ACE inhibitor plus B-blocker (n = 14; 28%) and B-blocker plus diuretic (n = 12; 24%)., Conclusion: The prevalence of hypertension in this patient population was low compared to the national average. This study shows that control of hypertension is not optimal, but higher than those reported elsewhere.

A DNA microarray chip of four virulence genes and 16S ribosomal DNA gene conserved region among all Gram negative species, including Yersinia, as a positive control was developed and evaluated using 22 Yersinia enterocolitica isolates. Eight different oligonucleotide probes (oligoprobes) with an average size of 22 bp, complementary to the unique sequences of each gene, were designed and immobilized on the surface of chemically modified slides. Multiplex PCR was used to simultaneously amplify DNA target regions of all five genes, and single stranded DNA (ssDNA) samples for microarray analysis were prepared by using a primer extension of amplicons in the presence of one primer of all genes. The presence of genes in Y. enterocolitica was established by hybridization of the fluorescently labeled ssDNA representing different samples of the microarray gene-specific oligoprobes and confirmed by PCR. Results of the study showed specificity of genotyping Y. enterocolitica using multiple microarray-based assays. Final validation of the chip's ability to identify Y. enterocolitica genes from adulterated pasteurized whole milk was confirmed and successful. The limit of chip detection of virulence genes in pasteurized whole milk was found to be 1000 CFU per hybridization.

A microarray-based method for characterization of six Clostridium perfringens toxin genes: iA (iota toxin), cpa (alpha toxin), cpe (enterotoxin E), etxD (epsilon toxin), cpb1 (beta toxin 1),and cpb2 (beta toxin 2) was developed and evaluated using 17 C. perfringens isolates. Three individual oligonucleotide probes (oligoprobes), complementary to the unique sequences of each toxin gene, were designed and immobilized on a surface of aldehyde-coated glass slides. Multiplex PCR was used to simultaneously amplify DNA target regions of all six genes. Single-stranded DNA (ssDNA) samples for microarray analysis were prepared by following a primer extension of amplicons in the presence of one primer. Fluorescent moieties (Cy3) were incorporated into the ssDNA by chemical modification of guanine bases. The presence of toxin genes in C. perfringens was established by hybridization of the fluorescently labeled ssDNA representing different samples to the microarray gene-specific oligoprobes. Results of the study showed sensitivity and specificity of genotyping C. perfringens using multiple microarray-based assays.

This study describes the application of filtration, infrared spectroscopy, and multivariate analysis to the identification of 10 foodborne bacterial species. The bacteria were applied by filtration to a disposable optical membrane that is transparent to infrared radiation. The filtration step was rapid (2 min). Observed cellular infrared spectra were unique and were used to discriminate among the different species. A dataset for the 10 bacterial species investigated was successfully used to correctly identify unknowns included in the dataset.

Six Selenomonas ruminantium strains (132c, JW13, SRK1, 179f, 5521c1, and 5934e), Streptococcus bovis JB1, and Bacteroides ovatus V975 were examined for nuclease activity as well as the ability to utilize nucleic acids, ribose, and 2-deoxyribose. Nuclease activity was detected in sonicated cells and culture supernatants for all bacteria except S. ruminantium JW13 and 179f sonicated cells. S. ruminantium strains were able to utilize several deoxyribonucleosides, while S. bovis JB1 and B. ovatus V975 showed little or no growth on all deoxyribonucleosides. When S. ruminantium strains 5934e, 132c, JW13, and SRK1 were incubated in medium that contained 15 mm ribose, the major end products were acetate, propionate, and lactate. S. ruminantium 5521c1 and S. bovis JB1 did not grow on ribose, and none of the S. ruminantium strains or S. bovis JB1 grew on 15 mm 2-deoxyribose. In contrast, B. ovatus V975 was able to grow on ribose and 2-deoxyribose. In conclusion, all S. ruminantium strains, S. bovis JB1, and B. ovatus V975 had nuclease activity. However, not all bacteria were able to utilize deoxyribonucleosides, ribose, or 2-deoxyribose.

To investigate the interaction between the manganese-stabilizing protein (MSP) and cytochrome c-550 (cyt. c-550) of the photosystem II (PSII) complex in the cyanobacterium Synechocystis sp. PCC6803, three site-directed amino acid substitution mutants in MSP (MSP-D159N, MSP-R163L, MSP-D 159N/R 163L) were created by single and double amino acid substitution mutagenesis. The modified psbO genes encoding the mutants forms of MSP were used to transform a single-deletion mutant deltapsO strain lacking MSP as well as a double-deletion strain deltapsbO:deltapsbV lacking both MSP and cyt. c-550. The mutant forms of MSP were expressed in each case and all permitted autotrophic growth in strains expressing cyt. c-550. However, when the MSP mutations were introduced into a strain which lacks cyt. c-550 (deltapsbV), the two single amino acid substitution mutants (deltapsbV:MSP-D159N and deltapsbV:MSP-R 163L) failed to grow photoautotrophically. These strains exhibited coupled O2-evolving activity of 68-77% compared to the wild-type control using CO2 as an electron acceptor and maximal uncoupled O2-evolution rates of 42-57% using 2,6-dichloro-p-benzoquinone (DCBQ) as an artificial electron acceptor. Interestingly, when the two amino acid substitutions were together in the absence of cyt. c-550 (deltapsbV:MSP-D159N/R163L), the mutant grew photoautotrophically and the oxygen-evolving activities were higher than in the single mutants. This indicates that the MSP-D159N mutant suppresses the non-autotrophic phenotype of MSP-R163L (or vice versa) in the absence of cyt. c-550. The possibilities of a direct (ionic) or indirect interaction between D159 and R163 of MSP are discussed.

The objective of this study was to evaluate the ability of the human colonic bacterium Bacteroides ovatus V975 to ferment fenugreek fiber (Fenufibers), psyllium husk (Metamucil), and wheat bran (Wheat Chex). Strain V975 was incubated in basal medium that contained 0.1 g of each fiber source for 0, 24, or 48 h. Little digestion of either fiber source was detected over 48 h, and little acetate or succinate was produced. From the lack of significant fiber digestion and fermentation by B. ovatus, it seems that all three fiber sources could be used as dietary supplements to increase roughage in the human diet.

A cryptic plasmid (pSR1) isolated from Selenomonas ruminantium HD4 was previously cloned into the HindIII site of pBR322 and a restriction map was constructed using HindIII, ClaI, BamHI, and PvuII (S. A. Martin and R. G. Dean, Appl. Environ. Microbiol. 55(12), 3035-3038, 1989). Analysis of the nucleotide sequence of pSR1 revealed two major open reading frames (ORFs) located in the minus strand at different frames. Analysis of ORF-1 revealed that it has 325 amino acids with a predicted MW of 36,588, and ORF-2 has 379 amino acids with a predicted MW of 42,651. The ORF-1 amino acids showed 30 to 32% sequence homology to the hypothetical protein YtqA in Bacillus subtilis and another hypothetical protein in the thermophilic bacterium Aquifex aeolicus. ORF-2 showed limited homology (23%) to the hypothetical protein ICFG in the photosynthetic cyanobacteria Synechocystis PCC6803., (Copyright 1999 Academic Press.)