Cleaving Folded RNA with DNAzyme Agents.

Cleavage of biological mRNA by DNAzymes (Dz) has been proposed as a variation of oligonucleotide gene therapy (OGT). The design of Dz-based OGT agents includes computational prediction of two RNA-binding arms with low affinity (melting temperatures (Tm ) close to the reaction temperature of 37 °C) to avoid product inhibition and maintain high specificity. However, RNA cleavage might be limited by the RNA binding step especially if the RNA is folded in secondary structures. This calls for the need for two high-affinity RNA-binding arms. In this study, we optimized 10-23 Dz-based OGT agents for cleavage of three RNA targets with different folding energies under multiple turnover conditions in 2 mM Mg2+ at 37 °C. Unexpectedly, one optimized Dz had each RNA-binding arm with a Tm ≥60 °C, without suffering from product inhibition or low selectivity. This phenomenon was explained by the folding of the RNA cleavage products into stable secondary structures. This result suggests that Dz with long (high affinity) RNA-binding arms should not be excluded from the candidate pool for OGT agents. Rather, analysis of the cleavage products' folding should be included in Dz selection algorithms. The Dz optimization workflow should include testing with folded rather than linear RNA substrates.

Marker-dependent cleavage of RNA by binary (split) DNAzyme (BiDz) and binary DNA machines (biDNM).

Oligonucleotide gene therapy (OGT) can be used to suppress specific RNA in cells and thus have been explored for gene therapy. Despite extensive effort, there is no clinically significant OGT for treating cancer. Low efficiency of OGT is one of the problems. Earlier, we proposed to address this problem by suppressing most vital genes in cancer cells e.g. housekeeping genes. To achieve specific activation of the OGT agents in cancer but not in normal cells, we designed binary (split) DNAzyme (BiDz), which is activated by cancer-related nucleic acid sequences. This work is devoted to BiDz optimization using cancer marker-related sequence as an activator and three folded RNA targets. To achieve efficient binding of folded RNA, the BiDz design was equipped with RNA binding/unwinding arms, a construction that was dabbed 'BiDz machines' (BiDM). BiDM was designed to have improved both iRNA cleavage rates and RNA recognition in comparison with BiDz. Further development of DNA nanotechnology-inspired agents can advance OGT technology in treating cancer, viral infections, and genetic disorders.

Cleaving Folded RNA by Multifunctional DNAzyme Nanomachines.

Both tight and specific binding of folded biological mRNA is required for gene silencing by oligonucleotide gene therapy agents. However, this is fundamentally impossible using the conventional oligonucleotide probes according to the affinity/specificity dilemma. This study addresses this problem for cleaving folded RNA by using multicomponent agents (dubbed 'DNA nanomachine' or DNM). DNMs bind RNA by four short RNA binding arms, which ensure tight and highly selective RNA binding. Along with the improved affinity, DNM maintain the high sequence selectivity of the conventional DNAzymes. DNM enabled up to 3-fold improvement in DNAzymes catalytic efficiency (kcat/Km) by facilitating both RNA substrate binding and product release steps of the catalytic cycle. This study demonstrates that multicomponent probes organized in sophisticated structures can help to achieve the balance between affinity and selectivity in recognizing folded RNA and thus creates a foundation for applying complex DNA nanostructures derived by DNA nanotechnology in gene therapy.

RNA-Cleaving DNA Thresholder Controlled by Concentrations of miRNA Cancer Marker.

Oligonucleotide gene therapy (OGT) agents suppress specific mRNAs in cells and thus reduce the expression of targeted genes. The ability to unambiguously distinguish cancer from healthy cells can solve the low selectivity problem of OGT agents. Cancer RNA markers are expressed in both healthy and cancer cells with a higher expression level in cancer cells. We have designed a DNA-based construct, named DNA thresholder (DTh) that cleaves targeted RNA only at high concentrations of cancer marker RNA and demonstrates low cleavage activity at low marker concentrations. The RNA-cleaving activity can be adjusted within one order of magnitude of the cancer marker RNA concentration by simply redesigning DTh. Importantly, DTh recognizes cancer marker RNA, while cleaving targeted RNA; this offers a possibility to suppress vital genes exclusively in cancer cells, thus triggering their death. DTh is a prototype of computation-inspired molecular device for controlling gene expression and cancer treatment.

Urinary biomarkers for early detection of platinum based drugs induced nephrotoxicity.

BACKGROUND: Nephrotoxicity is a major hazard complicating the use of platinum based drugs (PBD), which can hinder using higher doses protocols to maximize the therapeutic gain. Shortage of serum creatinine level as an accurate biomarker for acute kidney injuries (AKI) necessitates searching for novel biomarkers with better sensitivity and specificity in patients on PBD. METHODS: In a prospective cohort design, 132 patients receiving PBD were selected for the study. AKI was diagnosed by continuous follow up of serum creatinine level according to Kidney Disease: Improving Global Outcomes (KDIGO) guidelines 2012. Serum creatinine and urinary biomarkers (KIM-1, NGAL and cystatin C) was measured in the day of treatment and for 3 days after PBD cycle. RESULTS: AKI occurred in 35 patients (26.52% of patients). KIM-1, Cystatin C, and NGAL showed significant increase in samples collected in the day of AKI in comparison to their corresponding basal levels (P <  0.0001). In addition, significant increase in urinary levels of the biomarkers in samples collected 1 day before AKI in comparison to their basal levels (P <  0.0001, P <  0.0001, and P = 0.013 for KIM-1, NGAL and Cystatin C respectively). Furthermore KIM-1 data showed a significant increase 2 days before serum creatinine rise in comparison to the corresponding KIM-1 levels in patients who developed AKI (P = 0.001). CONCLUSIONS: Urinary KIM-1, Cystatin C and NGAL can predict PBD induced AKI in earlier stages than serum createnine. KIM-1 is the most sensitive biomarker for early detection of AKI in patients receiving PBD.

RNase H-dependent DNA thresholder modulated by cancer marker concentration.

Threshold antisense oligonucleotide constructs were designed to cleave mRNA within different biomarker concentrations. The mRNA cleavage is activated by 2.6, 7.5 or 39.5 nM of biomarker depending on the construct design. The constructs can be used to differentiate cancer from normal cells by the level of oncogene expression followed by silencing of a targeted gene.

Thresholding DNA Nanomachine as a Highly Cooperative and Efficient Enzyme-Like System for Controlled RNA Cleavage.

Therapeutic nucleic acid agents (TNA) can be activated by a marker RNA sequence followed by initiation of targeted RNA cleavage. This property can be used in conditional cell suppression, e. g., cancer marker-dependent cell death. However, healthy cells often express lower levels of cancer markers, thus jeopardizing TNA activation exclusively in cancer cells. Earlier, we developed a conditionally activated split deoxyribozyme construct (DNA thresholder or DTh) that can be activated by high but not by low concentrations of cancer markers. It's activity, however, was suppressed by very high marker concentrations. Herein, we combine the DTh functional units in a single DNA association (Thresholding DNA nanomachine or Th-DNM). Th-DNM maintains a high level of RNA cleavage activity in the presence of marker concentrations above the threshold level. Th-DNM differentiated fully complementary miR17 markers sequence from double base mismatched miR-20. Th-DNM can become a building block of DNA nanorobots for cancer treatment.

Effect of Ramadan fasting on chronic inflammation markers and gut bacterial endotoxins among Egyptian hemodialysis patients.

BACKGROUND: Chronic kidney disease (CKD) is associated with a state of chronic inflammation. This study aimed to investigate effects of Ramadan fasting on the markers of chronic inflammation and gut bacterial endotoxin levels in maintenance hemodialysis. METHOD: A prospective self-controlled observational study included 45 patients. Serum levels of High Sensitive CRP (hsCRP), indoxyl sulfate, and trimethylamine-n-levels were measured within a week before and a week after Ramadan fasting. RESULTS: Twenty-seven patients have fasted more than 15 days (29 ± 2.2 days). The levels of high sensitive C-reactive protein (hsCRP) (median of 62 mg/L vs. 91 mg/L), trimethylamine-n-oxide (TMAO) (median of 4.5 µmoL/L vs. 17 µmoL/L), platelet-to-lymphocyte ratio (PLR) (mean of 98.9 mg/L vs. 111.8 mg/L) and neutrophil-to-lymphocyte ratio (NLR) (median of 1.56 vs. 1.59) were significantly lower after Ramadan fasting with p < 0.001, p < 0.001, p < 0.001, and p = 0.04, respectively. CONCLUSION: A beneficial effect of Ramadan fasting on levels of bacterial endotoxins and markers of chronic inflammation in hemodialysis patients was observed.

Influence of time factor and albuminuria on characteristics of patients with type 2 diabetes Mellitus before, during and 1 year after COVID-19 recovery.

BACKGROUND: The potential effects of time factor and albuminuria on the morbid alterations in patients with type 2 diabetes (T2D) and COVID-19 are still unclear. We aimed to address the morbid alterations and the potential effects of time factor and albuminuria on the patients' characteristics before, during, and 1 year after COVID-19 recovery. METHODS: 83 patients with T2D were included, at Mansoura University Hospital, Egypt (July 2021-December 2021). Data of detailed history, physical examination, laboratory tests were recruited from files of the patients. Diagnosis and resolution of COVID-19 were established by Real time polymerase chain reaction (RT-PCR) test of SARS-CoV2. Complete blood count (CBC), renal and hepatic function tests, multiple measures of morning spot urine albumin to creatinine ratio (urine ACR), glycosylated hemoglobin (HBA1c), lipid profile, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), Ferritin, neutrophil to lymphocyte ratio (NLR), vitamin D3, intact parathyroid hormone (intact PTH), serum calcium were applied to all participants. RESULTS: Our participants had a mean age of 45 years, 60.2% male, 56.6% were hospitalized, and 25.3% were admitted to ICU for severe COVID-19. Albuminuria was prevalent in 71.1% before, 98.8% during, and 92.8% after COVID-19 recovery. Patients with albuminuria showed older age, longer duration of T2D, more frequent severe COVID-19 and hospitalization (p = 0.03, p < 0.001, p = 0.023& p = 0.025) respectively. Body mass index (BMI), mean arterial blood pressure, ESR, CRP, ferritin, NLR, HBA1c, triglycerides to high-density lipoprotein cholesterol (TG/HDL-C) ratio, vitamin D3, serum calcium, alkaline phosphatase (ALP), hepatic aminotransferases, and urine ACR showed significant alterations throughout the study (p < 0.001 for all). Although the interaction between time and albuminuria showed non-significant effect on all studied parameters, we noticed relevant main effects of time factor on Body mass index (BMI), HBA1c, glomerular filtration rate (eGFR), TG/HDL ratio, NLR, vitamin D3, (p < 0.001 for all). Moreover, albuminuria showed main effects on BMI, serum creatinine, and intact PTH (p = 0.019, 0.005 & <0.001), respectively. CONCLUSION: The characteristics of patients with T2D significantly altered throughout the study. Time factor and albuminuria exerted relevant main effects on the patients' characteristics without significant effect of their interaction.

Influence of sodium-glucose Co-transporter 2 inhibitors on clinical and biochemical markers of dehydration during the Holy Ramadan.

BACKGROUND AND AIMS: Alteration of the hydration status with the use of sodium-glucose co-transporter- 2 inhibitors (SGLT2i) during the Holy Ramadan has not been studied in depth. Precisely, we aimed to detect the potential alteration of hydration status in adult Muslims with type 2 diabetes mellitus (T2D) who used SGLT2i during Ramadan. METHODS: An observational non-interventional study included 245 patients with type 2 diabetes mellitus of matched age and sex. The study included 3 groups: empagliflozin group; 87 patients, dapagliflozin group; 85 patients and control group; 73 patients without the use of SGLT2i. Participants in each group were well-settled on their medications for more than 3 months before the onset of Ramadan. Clinical and biochemical parameters of hydration status were evaluated during the last week of Ramadan. RESULTS: We noticed a higher prevalence of orthostatic dizziness and postural hypotension among SGLT2i users than non-SGLT2i users (p < 0.001). The mean arterial blood pressure was significantly lowered among users of empagliflozin and dapagliflozin than non-SGLT2i users; 93.7 ± 5.1 and 93.1 ± 6.9 versus 106.2 ± 4.3, p < 0.001, respectively. Moreover, patients who used empagliflozin or dapagliflozin exhibited significantly higher values of urine specific gravity; 1029.6 ± 1.5 and 1029.1 ± 1.6 versus 1016.9 ± 4.4, p < 0.001, serum osmolality; 300.7 ± 10.2 and 297.8 ± 8.9 versus 290.9 ± 6.7, p < 0.001, and BUN/creatinine ratio; 24.1 ± 4.1 and 23.2 ± 4.6 versus 16.3 ± 4.2, p < 0.001 than non-SGLT2i users. CONCLUSION: Significant clinical and biochemical markers of dehydration were noticed among users of SGLT2i during the Holy Ramadan.

Binary Antisense Oligonucleotide Agent for Cancer Marker-Dependent Degradation of Targeted RNA.

Antisense oligonucleotide technology is one of the most successful gene therapy (GT) approaches. However, low selectivity of antisense agents limits their application as anticancer drugs. To achieve activation of antisense agent selectively in cancer cells, herein, we propose the concept of binary antisense oligonucleotide (biASO) agent. biASO recognizes an RNA sequence of a gene associated with cancer development (marker) and then activates RNase H-dependent cleavage of a targeted messenger RNA. biASO was optimized to produce only the background cleavage of the targeted RNA in the absence of the activator. The approach lays the foundation for the development of highly selective and efficient GT agents.

The Beneficial Impact of Intraoperative Ultrasound on Resection Margin Status during Breast Conserving Surgery.

Background: Surgical resection with clear surgical cut margins is the mainstay of managing malignant breast neoplasms. Multiple techniques have been suggested to enhance resection status during breast-conserving surgery (BCS), including intraoperative ultrasonography (IOUS). Herein, we conducted the current investigation to reveal the benefit of IOUS on the achievement of R0 resection. Patients and Methods. This retrospective investigation included 140 patients who underwent BCS. They were divided into two groups: the IOUS group (40 cases) and the control group (100 cases). Our primary objective was to determine the free resection margin status (R0). Results: Both study groups expressed statistically comparable demographic and clinical data. Additionally, histopathological examination revealed no significant difference between the two groups regarding the tumor type, stage, or grade. Nonetheless, the R0 resection margin was more frequently encountered in association with IOUS application (97.5% compared to 79% in the control group), and that difference was statistically significant (p=0.007). Conclusion: The application of IOUS has a significant beneficial impact on the outcomes of BCS. It is associated with a marked decline in positive resection margins, and its application should be encouraged in the breast oncological practice.

Controlling of Meloidgyne incognita (Tylenchida: Heteroderidae) using nematicides, Linum usitatissimum extract and certain organic acids on four peppers cultivars under greenhouse conditions.

Organic acids and plant extracts, which have a nemacidal action and may be used instead of nematicides that pollute the environment, are one way for controlling the pepper root-knot nematode. We provide in this study for a first time a new strategy for management Meloidgyne incognita (Kofoid and White) by using organic acids and plant extract compared to nematicides on four peppers cultivars (Super amarr, Super mard, Super noura and Werta) under greenhouse conditions compared to nematicides. This study aimed to evaluate 0.1% of organic acids (humic and salicylic acid) and 0.1% of Linum usitatissimum extract on plant parameters of pepper varieties (Super amarr, Super mard, Super noura and Werta) and control of M. incognita under greenhouse conditions compared to four nematicides (Oxamyl 24% SL, Fosthiazates 75% EC, Ethoprophos N40% EC and Fenamiphos 40% EC). Our data obtained four nematicides were more effectiveness than other treatments in reduced galls and egg masses of M. incognita. Whilst, humic and salicylic acids have remarkably higher nematicidal activity than L. usitatissimum in all lines of pepper. Therefore, plant extract and organic acids may be used a best alternative of nematicides to control PPNs and caused the longitudinal growth of plant. Also, ultimately reduce environmental risk from nematicide pollution.

Mite pests and their predators on seven vegetable crops (Arachnida: Acari).

During this study the frequency of occurrence and dominance of phytophagous and predatory mites harboring seven vegetable crops in Egypt, namely common bean, cowpea, eggplant, okra, squash, sweet pepper and sweet potato during 2017-2018 were investigated to identify predatory mites that might be useful for the biological control of the phytophagous mites. Three phytophagous and nine predatory mite species were surveyed. The two spotted spider mite Tetranychus urticae Koch of the family Tetranychidae was the dominant pest on these vegetables, while phytoseiids Phytoseiulus persimilis (Athias- Henriot), Typhlodromips swirskii (Athias- Henriot) and Euseius scutalis Chant were the dominant predators. The population of the native or indigenous phytoseiid mite fauna in Egypt such as Phytoseiulus persimilis could be considered as a good biocontrol agent and a part of the Integrated Pest Management (IPM) program in the future. Mite fauna of Egypt especially local populations of Phytoseiulus persimilis can be considered for implementation in future Integrated Pest Management (IPM).

A Flexible Extension of Pareto Distribution: Properties and Applications.

This paper introduced a relatively new mixture distribution that results from a mixture of Fréchet-Weibull and Pareto distributions. Some properties of the new statistical model were derived, such as moments with their related measures, moment generating function, mean residual life function, and mean deviation. Furthermore , different estimation methods were introduced for determining the unknown parameters of the proposed model. Finally, we introduced three real data sets which were applied to our distribution and compared them with other well-known statistical competitive models to show the superiority of our model for fitting the three real data sets, and we can clearly see that our distribution outperforms its competitors. Also, to verify our results, we carried out the existence and uniqueness test to the log-likelihood to determine whether the roots are global maximum or not.

In vivo and in vitro management of Meloidogyne incognita (Tylenchida: Heteroderidae) using rhizosphere bacteria, Pseudomonas spp. and Serratia spp. compared with oxamyl.

Biological control using rhizosphere bacteria, Pseudomonas spp. and Serratia spp. is a prospective alternative technique to overcome plant parasitic nematodes infection. So, the current study was conducted in vitro on five egg-masses, 100 free eggs and 100 infective juveniles (IJs) of Meloidogyne incognita as well as greenhouse treatments on Luffa aegyptiaca L. to evaluate the nematicidal potential of six strains belong to Pseudomonas spp. and Serratia spp. as compared to oxamyl. Results showed that the inhibitory effect and juvenile mortality varied according to bacteria species, strains and exposure time. All the tested bacteria significantly (P ≤ 0.05) inhibited egg hatching and increased juvenile mortality in vitro. After 3 days of treatment, Pseudomonas spp. were more effective against eggs (48.31to 55.15%) and IJs (20.98 to 25.30%) than Serratia spp. (44.55 to 49.75% with eggs) and (19.06 to 21.61% with IJs), respectively. In the pot experiment, Luffa aegyptiaca L. treated with Serratia spp. and Pseudomonas spp. displayed significantly higher (P ≤ 0.05) levels of growth (as indicated by root length, fresh roots weight and fresh shoots weight) compared to control plants and significantly (P ≤ 0.05) suppressed galling (number of galls) and reproduction (as indicated by number of egg-masses on roots and number of eggs and juveniles in pot soil). Meanwhile, among the treated plants, Serratia spp. and Pseudomonas spp. gave the best results in shoot weight of pots infected by eggs of M. incognita than those infected with IJs as compared with positive control. While, oxamyl treatment gave the best results in pots infected by eggs and IJs. The lowest galling (gall index), number of eggs and juveniles in soil was observed in the treatment with mixture of Serratia spp. and Pseudomonas spp. as well as, enhanced growth of sponge gourd more than application each of them alone. Pots treated with oxamyl overwhelmed those treated with mixture of Serratia spp. and Pseudomonas spp.

Effect of Ramadan fasting on arterial stiffness parameters among Egyptian hypertensive patients with and without chronic kidney disease.

Arterial stiffness (AS) increases progressively in patients with chronic kidney disease (CKD). It is a strong predictor of cardiovascular and all-cause mortality. This study aims at evaluate of the effect of Ramadan fasting on AS parameters, augmentation index (AIx), and pulse wave velocity (PWV) in hypertensive patients with and without CKD. A cohort of 71 patients (mean age = 57.14 ± 14.5 years, 42 females and 29 males) were enrolled in this study; 34 with CKD and 37 without CKD. All patients had hypertension, while 25 patients had diabetes mellitus. Serum creatinine (Cr), serum urea, estimated glomerular filtration rate (eGFR) by CKD-EPI formula, brachial and central systolic blood pressure (BSP and CSP respectively), brachial and central diastolic blood pressure (BDP, CDP, respectively), AIx and PWV (measured by cuff based oscillometric method) were assessed before and after Ramadan fasting. In patients without CKD BSP, BDP, CSP, and CDP significantly decreased (P = 0.0001, 0.0001, 0.0001, and 0.0001, respectively). In patients with CKD BSP and CSP significantly decreased (P = 0.005 and 0.005), while BDP and CDP decreased, but the change was not statistically significant. AIx significantly decreased in patients without CKD (P = 0.0001, mean 36.24 before and 26.22 after Ramadan fasting), but did not significantly change in patients with CKD (P 0.381 mean 25.94 before and 25 after Ramadan fasting). PWV decreased in both groups, but the change was not significant. Serum Cr significantly increased (P = 0.03 mean 1.06 mg/dL before and 1.11 mg/dL after Ramadan fasting), while eGFR did not significantly decrease (P = 0.072, mean 69.73 mL/ min/1.73 m2 before and 67.3 mL/min/1.73 m2 after Ramadan fasting) in patients without CKD. Serum Cr significantly decreased (P 0.028 mean 1.93 mg/dL before and 1.87 mg/dL after Ramadan fasting) and eGFR significantly increased (P 0.006 mean 32.65 mL/min/1.73 m2 before and 34.68 mL/min/1.73 m2 after RF) in patients with CKD. Ramadan fasting is associated with improved peripheral and central blood pressure control in hypertensive patients with and without CKD. It is also associated with improved arterial compliance (decreased AIx) in hypertensive patients without CKD.

Insulin Resistance and Hepatitis C Virus-Associated Subclinical Inflammation Are Hidden Causes of Pruritus in Egyptian Hemodialysis Patients: A Multicenter Prospective Observational Study.

BACKGROUND: Pruritus is one of the most common and disturbing symptoms in hemodialysis (HD) patients. The pathogenesis of pruritus in HD patients is multifactorial; however, a little progress in understanding this pathogenesis has been achieved. OBJECTIVES: To evaluate the frequency and risk factors of pruritus among HD patients in Dakahlia, Egypt. METHODS: A total of 193 patients from 4 HD centers were included in this study. Pruritus intensity was assessed by visual analog scale. All patients were assessed by clinical and laboratory parameters, such as age, sex, duration of dialysis, dialysis adequacy, urea, calcium, phosphorus, parathyroid hormone, fasting insulin, fasting sugar, HOMA-IR, serum ferritin, high-sensitive C-reactive protein (hs-CRP) and hemoglobin. They were also investigated for hepatitis C virus (HCV) infection by HCV enzyme-linked immunosorbent assay, HCV-PCR for plasma and buffy coat for further detection of occult HCV infection. RESULTS: Male gender, dialysis duration, inadequate dialysis, anemia, high ferritin, hyperphosphatemia, hypocalcemia, hs-CRP, and insulin resistance were characteristic features in pruritic patients (p = 0.01, 0.006, 0.0001, 0.047, 0.01, 0.0001, 0.024, 0.000, and 0.0001, respectively). Significant positive correlations were found between pruritus score and each of age (p = 0.002, r = 0.222), duration of dialysis (p = 0.03, r = 0.151), serum ferritin (p = 0.001, r = 0.213), serum phosphorus (p = 0.0001, r = 0.59), fasting insulin (p = 0.001, r= 0.273), and HOMA-IR (p = 0.0001, r = 0.349), while there was a negative correlation with Kt/V (p= 0.0001, r = -0.459). Linear multivariate regression analysis showed that age, duration of dialysis, serum phosphorus, Kt/v, and hs-CRP were good predictors for pruritic score in HD patients. All HCV-infected patients (who were positive for both plasma and buffy coat HCV-PCR) had pruritus with -significantly higher pruritus score than non-infected patients (p = 0.009), they also showed significantly higher fasting insulin, HOMA-IR, and hs-CRP levels (p = 0.0001). CONCLUSIONS: Uremic pruritus (UP) is a serious problem in HD patients. hs-CRP, male gender, dialysis duration, insulin resistance, dialysis inadequacy, and hyperphosphatemia are positively correlated with the intensity of UP. HCV infection is associated with severe UP, insulin resistance, and inflammation.

Homocysteine: a new diagnostic marker in spontaneous bacterial peritonitis.

BACKGROUND AND AIMS: The diagnosis of spontaneous bacterial peritonitis (SBP) depends primarily on a polymorphonuclear leukocyte cell count more than 250/mm. This method is invasive, and not diagnostic in all variants of SBP; we aimed to assess serum homocysteine as a precise indicative marker for the diagnosis of all variants of SBP. PATIENTS AND METHODS: A total 323 consecutive ascitic patients were registered in this prospective work. Serum and ascitic fluid of homocysteine were evaluated utilizing an enzyme-linked immunosorbent assay. RESULTS: Participants were classified into a non-SBP group, including 262 participants and 61 patients with SBP. Serum and ascitic homocysteine were considerably elevated in the SBP group than in the non-SBP group (17.94±7.57 vs. 11.75±5.68 µmol/l; P<0.001 and 14.70±5.45 vs. 9.75±4.55 µmol/l; P<0.001). At a cutoff value of 17.79 µmol/l, serum homocysteine had 89.3% specificity and 95.1% sensitivity for distinguishing SBP (area under the curve: 0.932) and, at a cutoff value of 16.1 µmol/l, ascitic homocysteine had 84.4% specificity and 92.7% sensitivity for distinguishing SBP (area under the curve: 0.901). Both were positively correlated with the polymorphonuclear count, C-reactive protein, Child-Pugh score, and Model For End-Stage Liver Disease score as well as negatively correlated with the protein content in the ascitic fluid and estimated glomerular filtration rate. After SBP therapy, there was a marked reduction in serum and ascitic homocysteine levels. CONCLUSION: This study demonstrates that serum and ascitic homocysteine are considerably higher in SBP participants versus non-SBP patients. Serum homocysteine may provide a reliable and noninvasive diagnostic marker for all variants of SBP.

Rifaximin in nonalcoholic fatty liver disease: hit multiple targets with a single shot.

BACKGROUND/AIMS: The pathogenesis of nonalcoholic fatty liver disease (NAFLD) may include increased insulin resistance, upregulation of proinflammatory cytokines, lipopolysaccharide, and BMI. Rifaximin is a minimally absorbable antibiotic that might act against a broad spectrum of gut bacteria. This study aimed to investigate the effects of rifaximin on NAFLD. PATIENTS AND METHODS: Fifty participants with biopsy-proven nonalcoholic steatohepatitis (NASH) were registered in this multicentric, double-blind, randomized, placebo-controlled study. BMI, alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transferase, lipid profile, serum endotoxin, homeostatic model assessment, toll-like receptor-4, interleukin-10 (IL-10), IL-6, tumor necrosis factor-α, and cytokeratin-18 (CK-18) levels were evaluated at baseline and at 1, 3, and 6 months of rifaximin therapy (1100 mg/day). RESULTS: Patients were randomized into two groups (rifaximin group; n=25 and placebo group; n=25). After 6 months of rifaximin therapy, patients with NASH showed a significant reduction in homeostatic model assessment, alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transferase, endotoxin, toll-like receptor-4, IL-6, tumor necrosis factor-α, CK-18, and NAFLD-liver fat score (all P<0.05), but no changes in the lipid profile; moreover, there was a mild nonstatistically significant reduction of BMI. However, in the placebo group, there was no significant difference in these variables at baseline and after therapy. CONCLUSION: Rifaximin therapy appears to be effective and safe in modifying NASH through reduction of serum endotoxin and improvement of insulin resistance, proinflammatory cytokines, CK-18, and NAFLD-liver fat score.