The potential therapeutic effect of metformin in type 2 diabetic patients with severe COVID-19.

OBJECTIVE: Type 2 diabetes mellitus (T2DM) is regarded as a chief risk factor for(coronavirus disease 2019 (COVID-19) owing to dysregulation of the expression of angiotensin-converting enzyme 2 (ACE2) and chronic low-grade inflammatory disorders. Metformin, an insulin-sensitizing agent for managing T2DM, has pleiotropic anti-inflammatory and oxidant potentials, which may lessen the risk of diabetic complications. So, we aimed to reveal the potential role of metformin monotherapy in treating T2DM patients with COVID-19. PATIENTS AND METHODS: In this prospective cohort study, 60 hospitalized T2DM patients with COVID-19 on metformin plus standard anti-COVID-19 treatments compared to 40 hospitalized T2DM patients with COVID-19 on other diabetic pharmacotherapy like insulin and sulfonylurea, were recruited. Inflammatory and oxidative stress biomarkers and radiological and clinical outcomes were assessed at admission time and at the time of discharge. RESULTS: The results of this study illustrated that metformin treatment in T2DM patients with COVID-19 was more effective in reducing inflammatory and oxidative stress biomarkers with significant amelioration of radiological scores and clinical outcomes compared to T2DM patients with COVID-19 on another diabetic pharmacotherapy. CONCLUSIONS: Our findings highlighted that metformin efficiently managed T2DM patients with COVID-19 by reducing inflammatory and oxidative stress with mitigating effects on the radiological scores and clinical outcomes.

Effect of Vitamin D supplementation in patients with liver cirrhosis having spontaneous bacterial peritonitis: a randomized controlled study.

OBJECTIVE: Spontaneous Bacterial Peritonitis (SBP) is one of the most serious liver cirrhosis with ascites complications. Vitamin D (Vit D) deficiency has been associated with a high risk of infection and mortality in cirrhotic patients. Herein, the assessment of Vit D level as a prognostic marker in SBP patients and the impact of Vit D supplementation on their treatment plan was studied as well. PATIENTS AND METHODS: Ascetic patients with SBP and Vit D deficiency were divided randomly into treatment and control groups. The control group received standard treatment without Vit D and the treatment group received standard treatment plus Vit D. Clinical monitoring of Vit D was done over 6 months. RESULTS: At baseline, all patients in both groups revealed an elevated serum and ascetic TLC, AST, ALT, total and direct bilirubin, in addition to elevation in INR and procalcitonin (PCT) level. Univariate regression analysis confirmed that deficiency of Vit D was an independent predictor of infection and mortality (p < 0.01; Crude Hazard Ratio: 0.951). Over 6 months, the study revealed significant improvement in serum Vit D level in the treatment group (34.6 ± 9.2 and 18.3 ± 10.0 ng/mL; p < 0.001). Moreover, a statistically significant increase in survival rate (64% vs. 42%; p < 0.05) and duration (199.5 days vs. 185.5 days; p < 0.05) were recorded as well. Univariate and multivariate regression analysis confirmed that Vit D supplementation was positively correlated to survival over 6 months (p < 0.001; Adjusted Hazard Ratio: 0.895). CONCLUSIONS: Vit D deficiency is prevalent in SBP cirrhotic patients and is used as an independent predictor of infection and death. Therefore, Vit D supplementation revealed improvement in their response to treatment.

Serotypes of Streptococcus pneumoniae in Egyptian children: are they covered by pneumococcal conjugate vaccines?

In Egypt, pneumococcal vaccines have not yet been introduced as being compulsory. Identification of the circulating serotypes in Egypt is mandatory to determine whether or not the pneumococcal vaccines will be beneficial. The current study aims to identify the serotypes, vaccine coverage, and antimicrobial resistance of Streptococcus pneumoniae colonizing the nasopharynx of Egyptian children younger than 5 years old. The study was conducted in two successive winter seasons (December 2012-February 2013 and December 2013-February 2014). Two hundred children were enrolled, aged from 6 months to 5 years, excluding those with fever, signs of infection, history of antibiotic intake, and hospitalization in the preceding month. Nasopharyngeal (NP) secretions were collected, subjected to culture, and underwent antibiotic susceptibility testing if positive for pneumococci. Real-time polymerase chain reaction (PCR) and serotyping by sequential multiplex PCR for positive cases were included as well. Streptococcus pneumoniae was isolated from 62 subjects. All isolates were sensitive to vancomycin and levofloxacin, but the majority showed resistance to multiple antibiotics. PCR was positive for pneumococci in 113 subjects (56.5%). The most commonly detected serotypes (st) were 6A6B6C (n = 21, 20.8%), 19F (n = 19, 18.8%), 1 (n = 11, 10.9%), 34 (n = 8, 7.9%), and 19A (n = 6, 5.9%). The theoretical coverage of the PCV13 vaccine for the detected serotypes was 72.4%, while that of PCV10 was 65.5%. Based on these percentages, we recommend including pneumococcal conjugate vaccines in the Egyptian national vaccination program.

The use of multiplex PCR for the diagnosis of viral severe acute respiratory infection in children: a high rate of co-detection during the winter season.

Respiratory tract infection is a major cause of hospitalization in children. Although most such infections are viral in origin, it is difficult to differentiate bacterial and viral infections, as the clinical symptoms are similar. Multiplex polymerase chain reaction (PCR) methods allow testing for multiple pathogens simultaneously and are, therefore, gaining interest. This prospective case-control study was conducted from October 2013 to February 2014. Nasopharyngeal (NP) and oropharyngeal (throat) swabs were obtained from children admitted with severe acute respiratory infection (SARI) at a tertiary hospital. A control group of 40 asymptomatic children was included. Testing for 16 viruses was done by real-time multiplex PCR. Multiplex PCR detected a viral pathogen in 159/177 (89.9 %) patients admitted with SARI. There was a high rate of co-infection (46.9 %). Dual detections were observed in 64 (36.2 %), triple detections in 17 (9.6 %), and quadruple detections in 2 (1.1 %) of 177 samples. Seventy-eight patients required intensive care unit (ICU) admission, of whom 28 (35.8 %) had co-infection with multiple viruses. AdV, HBoV, HRV, HEV, and HCoV-OC43 were also detected among asymptomatic children. This study confirms the high rate of detection of viral nucleic acids by multiplex PCR among hospitalized children admitted with SARI, as well as the high rate of co-detection of multiple viruses. AdV, HBoV, HRV, HEV, and HCoV-OC43 were also detected in asymptomatic children, resulting in challenges in clinical interpretation. Studies are required to provide quantitative conclusions that will facilitate clinical interpretation and application of the results in the clinical setting.

Yogurt containing bioactive molecules produced by Lactobacillus acidophilus La-5 exerts a protective effect against enterohemorrhagic Escherichia coli in mice.

An active fraction extracted from Lactobacillus acidophilus La5 cell-free spent medium (LAla-5AF) was incorporated in a dairy matrix and tested to assess its antivirulent effect against enterohemorrhagic Escherichia coli (EHEC). Mice in experimental groups were fed for 4 days with yogurt supplemented with LAla-5AF. On the fifth day, mice were challenged with a single dose (10(7) CFU per mouse) of E. coli O157:H7. The clinical manifestations of the infection were significantly less severe in mice fed the yogurt supplemented with LAla-5AF. EHEC attachment and colonization was attenuated by LAla-5AF. Tumor necrosis factor alpha production was down-regulated, which might indicate a protective effect in the kidney during EHEC infection. To investigate the mechanisms associated with the in vivo effects observed, LAla-5AF was tested by reverse transcription real-time PCR to confirm its effects on the expression of several virulence genes of EHEC O157. The results showed that these fractions were able to down-regulate several virulence genes of EHEC, including stxB2, qseA, luxS, tir, ler, eaeA, and hlyB.

Diagnosis of human brucellosis in Egypt by polymerase chain reaction.

This study aimed to establish an accurate and sensitive polymerase chain reaction (PCR) technique for the diagnosis of active human brucellosis in Egypt. We failed to extract Brucella DNA with a commercial kit, but an extraction kit designed in-house using 2 sets of primers [B4/B5 (223 bp) and JPF/JPR (193 bp)] was successful and more economical. The technique showed high sensitivity, specificity and accuracy. The PCR positivity increased significantly with increasing seropositivity titres by the standard tube agglutination test and showed 100% positivity in patients with positive blood cultures. We recommend using PCR as an alternative to culture for diagnosis of brucellosis.

Diagnosis of human brucellosis in Egypt by polymerase chain reaction

This study aimed to establish an accurate and sensitive polymerase chain reaction [PCR] technique for the diagnosis of active human brucellosis in Egypt. We failed to extract Brucella DNA with a commercial kit, but an extraction kit designed in-house using 2 sets of primers [B4/B5 [223 bp] and JPF/JPR [193 bp]] was successful and more economical. The technique showed high sensitivity, specificity and accuracy. The PCR positivity increased significantly with increasing seropositivity titres by the standard tube agglutination test and showed 100% positivity in patients with positive blood cultures. We recommend using PCR as an alternative to culture for diagnosis of brucellosis

Genetic characterisation of the Egyptian vaccinal strain Abu-Hammad of bovine herpesvirus-1.

The Egyptian Abu-Hammad vaccinal strain of bovine herpesvirus-1 (BHV-1) was genetically characterised by comparing HindIII endonuclease genomic fingerprints of the Egyptian BHV-1 with reference strain Cooper 1 of BHV-1 subtype 1 (BHV-1.1). Analyses of nucleotide (nt) and deduced amino acid (aa) sequences and phylogeny of the major viral immunogen, glycoprotein D (gD), were used to compare the Egyptian BHV-1 with related alphaherpesviruses. HindIII restriction digests revealed close identity between the Egyptian BHV-1 and reference BHV-1.1. Both nt and aa sequence alignments revealed variable degrees of sequence similarity with other alphaherpesviruses. Possible mutational frameshifts were observed at nt 509 and 615 of the Egyptian BHV-1 gD. The Egyptian vaccinal BHV-1 was grouped with BHV-1.1 in a distinct branch of the phylogenetic tree. Conservation of five cysteine residues and glycosylation domains emphasised the importance of the amino terminus for immunological and biological function of alphaherpesvirus gD. The most divergent domain of 17 residues at positions 168-184 and an additional cysteine residue at position 178 distinguish the Egyptian BHV-1 from other herpesviruses. This work demonstrated that HindIII genomic fingerprinting and sequencing of the gD gene are useful for genetic characterisation of BHV-1. They may also be applied to epidemiological studies and development of BHV-1 vaccines.

Use of polymerase chain reaction to simultaneously detect and type bovine viral diarrhoea viruses isolated from clinical specimens.

The techniques of indirect immunofluorescence (IF), immuno-peroxidase (IP) staining and the one-step reverse transcriptase polymerase chain reaction (RT-PCR) were compared for detection of 102 isolates of bovine viral diarrhoea virus (BVDV) in infected cell cultures. The BVDV was obtained from bovine clinical specimens, including sera, buffy coats and tissues, submitted from farms located in the States of Iowa and Wisconsin, United States of America. The IF technique detected 88/102 (86.3%) of the viral isolates, whereas IP staining detected an additional 4 isolates (92/102; 90%). The one-step RT-PCR using primers derived from the 5' untranslated region of the BVDV genome detected 102/102 (100%) of the BVDV isolates. A second-round PCR utilising another pair of PCR primers from the 5' untranslated region, allowed rapid genotyping of BVDV. The procedure used showed that the PCR assay based on the 5' untranslated region of the virus genome is the most sensitive indicator for BVDV detection in cell culture, and is also of considerable epidemiological importance since it allowed rapid genotyping of BVDV isolated from clinical specimens. In addition to detection and genotyping of BVDV isolated from clinical specimens, the RT-PCR procedure can be used for routine screening of locally produced and imported biologicals for BVDV contamination. However, the procedure requires further refinement to enable direct application on the clinical specimen.

Septal mucoperichondrial flap for closure of nostril in atrophic rhinitis.

The aetiology of primary atrophic rhinitis is still unknown. Treatment of this disease is conservative in the first place. Surgery is indicated if the medical treatment fails. The aim of surgery is either to narrow the nasal cavity or in special cases to close the nostril. Closure of the nostril (Young's operation), is achieved by raising a circular skin flap. Raising the skin flap is difficult, the suture line may break down and an excessive scar tissue may form resulting in vestibular stenosis. We therefore developed a septal mucoperichondrial flap to close the nostril. This new and easy technique has been used to treat 17 patients with excellent results. The description of this technique and the results of surgery will be discussed.

Arterial and venous paragangliomas: the value of a preoperative pacemaker and a multidisciplinary approach.

Non-chromaffin paragangliomas are very rare tumours. They arise from chemoreceptor cells at multiple sites throughout the body. They are usually very vascular and most arise in intimate contact with vital structures making their excision a surgical challenge. We describe in this paper four cases of paragangliomas in the carotid body and the jugular bulb. Their presentation was unusual. Two cases underwent excisional surgery and two had radiation treatment because they were not fit for surgery. We had to use a temporary pacemaker for preoperative preparation of the carotid body because of an associated hypersensitive carotid sinus syndrome. The clinical presentation and the surgical management will be described in this paper.

Vocal cord dysfunction after cardiac surgery: an overlooked complication.

OBJECTIVE: To evaluate the complication of vocal cord dysfunction following open heart surgery and its short- and long-term effects. MATERIALS AND METHODS: Five cases (1.9%) of vocal cord dysfunction which were diagnosed among 270 cases of adult open heart surgery performed at this centre between February 1993 and May 1995. RESULTS: All five patients developed respiratory insufficiency following tracheal extubation. They required reintubation and ventilation. Diagnosis was delayed in three cases because of unawareness of the problem. Tracheostomy was performed in three cases and minitracheostomy in two. In one case, shortness of breath on exertion persisted due to partial laryngeal obstruction from bilateral cord paresis and required arytenoidectomy 11 months postoperatively. CONCLUSIONS: Vocal cord dysfunction can be an easily overlooked complication after open heart surgery. It can be the cause of respiratory insufficiency following tracheal extubation and may lead to reintubation and reventilation. The cause of the problem cannot always be traced but it may be due to direct trauma of the vocal cords during tracheal intubation, or trauma of the recurrent laryngeal nerve from the cuff of the endotracheal tube. A less likely possibility is that it may result from nerve injury due to central venous cannulation, or from cold. The condition may resolve within months, but, in rare cases, may lead to permanent morbidity.

Rheumatic fever prophylaxis using benzathine penicillin G (BPG): two- week versus four-week regimens: comparison of two brands of BPG.

OBJECTIVE: This prospective study was aimed at answering two important questions: 1) Is a biweekly schedule of 1.2 million U intramuscular benzathine penicillin G (BPG) superior to a 4-week one in the prevention of upper respiratory Group A beta-hemolytic streptococcal (GABHS) infections and rheumatic fever (RF) recurrences? 2) Is there a difference in the bioavailability of BPG obtained from different manufacturers? METHODOLOGY: Three hundred sixty rheumatic patients aged 4 to 20 years were randomly assigned to either a biweekly (190 patients) or 4-week (160 patients) BPG prophylactic schedule and were followed-up monthly for 2 years by clinical examination, throat swab culture for GABHS and measurement of antistreptolysin O titer to detect GABHS infection and/or recurrences of RF (according to revised Jones' Criteria). Thereafter, 34 rheumatic subjects, aged 8 to 16 years were randomly assigned to receive a 4-week injection of 1.2 million U of either a locally manufactured BPG brand (22 patients) or an imported one (12 patients). Sera of all patients were tested for penicillin level by plate diffusion method on days 1, 2, 3, 4, 5, 6, 7, 14, 21, and 28 after the intramuscular injection of BPG. RESULTS: The GABHS infection rate was found to be 0.2% and 0.3% for patients on the biweekly and 4-week BPG schedules, respectively, with no significant differences between them. However, the RF recurrence rate/patient/year for the 4-week schedule patients (0.12) was double that for the biweekly schedule ones (0.06). Estimation of the bioavailability of the two different brands of BPG demonstrated a difference in their pharmacokinetics and a decrease in the serum penicillin concentration below the minimum inhibitory concentration 3 weeks after the injection of either brand. CONCLUSION: Although a biweekly schedule may not be superior in preventing upper respiratory GABHS infection, it may play a role in preventing the sequelae of such infections. The short duration of penicillinemia explains the superiority of the 2-week schedule in RF prophylais. The difference in the pharmacokinetics of penicillin brands might contribute to the high recurrence rate of RF reported in Egypt.

Cloning and expression of the nucleoprotein of peste des petits ruminants virus in baculovirus for use in serological diagnosis.

Peste des petits ruminants (PPR) is a viral disease of goats and sheep characterized by erosive stomatitis, enteritis, and pneumonia. The virus is a member of the family Paramyxoviridae and the genus Morbillivirus. The disease has high morbidity and mortality rates and has a substantial economic impact in developing countries. We have cloned and sequenced the cDNA of the nucleocapsid (N) gene of the Nigeria 75/1 strain of PPR virus (PPRV). A comparison of its nucleotide and deduced amino acid sequence with those of the N gene of the tissue culture-attenuated strain of PPRV was performed. A divergence of 8.9 and 5.0% was found at the nucleotide and amino acid level, respectively. A recombinant baculovirus that expresses the N protein in insect cells and larvae (Spodoptera frugiperda) was generated. The recombinant protein, characterized by Western blot analysis, was shown to have a molecular weight of 58 kDa and was recognized by anti-PPRV antibodies. The recombinant protein was used successfully as a coating antigen in an enzyme-linked immunosorbent assay for the serological diagnosis of PPRV.

Alterations of plasma free amino acids in nephrotic syndrome.

In order to assess the plasma amino acid pattern in nephrotics, we studied 19 nephrotic patients in good general and nutritional conditions. The results were compared with those of 8 healthy controls of matched age and sex. A highly significant rise of plasma glutamic acid and hydroxyproline was detected in nephrotics. When the latter group is divided into 2 subgroups according to the serum creatinine level, the rise in plasma values of both amino acids was detected in both subgroups compared to the controls, while an insignificant rise was detected in cases with elevated serum creatinine compared to those with normal serum level. So we conclude that there is an alteration in the amino acid pattern in nephrotics, and that this alteration occurs early, before the rise in serum creatinine.

Comparative study of distribution and biochemical characterization of brain calcitonin gene-related peptide receptors in five different species.

Calcitonin gene-related peptide receptors are abundant in the brain, and the pattern of distribution of receptors is similar in the rat, pig, cow, sheep and in man. In comparison to the cerebellum (100%), only a small number of receptors were found in the ventral spinal cord (20%), cerebral white matter (20%), pituitary (27%) and hypothalamus (33%). In contrast, cerebral cortex (70%), thalamus (50%) and dorsal spinal cord (50%) contained a higher number of calcitonin gene-related peptide receptors. Overall, the highest number of receptors was observed in the pig brain followed by the rat, human, sheep and cow. IC50 values of alpha- and beta-calcitonin gene-related peptide for cerebellar membranes were 200 pM, while its antagonist calcitonin gene-related peptide(8-37) and (9-37) had an IC50 approximately 1 nM. The recently discovered 37-amino acid peptide amylin (46% homology with calcitonin gene-related peptide) displaces the membrane bound [125I]calcitonin gene-related peptide with 50-fold molar excess (IC50 = 10 nM). The only other peptide able to compete for calcitonin gene-related peptide receptor binding is salmon calcitonin, at a > 1000-fold molar excess (IC50 = 250 nM). Dissociation of [125I]calcitonin gene-related peptide from cerebellar membranes was biphasic, suggesting that calcitonin gene-related peptide receptors in the brain were heterogeneous.(ABSTRACT TRUNCATED AT 250 WORDS)

Regulation of antigen-induced blastogenesis and interleukin-2 expression by adherent mononuclear cells in humans infected with Schistosoma mansoni.

The immune response and its regulation by adherent mononuclear cells were investigated in 39 Egyptians aged 13-30 (mean +/- SD, 16.2 +/- 3.6) with chronic Schistosoma mansoni infection (fecal egg excretion, 40-2480 eggs/g; mean, 709 +/- 612). Blastogenesis and production of interleukin-2 (IL-2) by peripheral blood mononuclear cells showed a significant correlation when streptolysin O but not when soluble worm antigenic preparation (SWAP) was used as stimulus. This suggested independent regulation of these responses to SWAP. Thirteen subjects showed SWAP-restricted suppression of blastogenesis but not IL-2 production by adherent cells. Compared with 24 other S. mansoni-infected subjects, they had depressed blastogenic responses to SWAP but comparable SWAP-induced IL-2 production and responsiveness to exogenous IL-2. This study indicates that IL-2 production is not the site of action of parasite antigen-restricted suppressor adherent cells in chronic S. mansoni infection.

Dissociation of immunoregulatory function of blood monocytes from maturational state and expression of interleukin-1 in humans chronically infected with Schistosoma mansoni.

We investigated the immunoregulatory function and properties of monocytes in 54 S. mansoni infected Egyptians (40-3,840 S. mansoni eggs excreted per gram of stool) 13-35 years of age. Adherent cell-mediated suppression was found in 11 of 36 patients. Cytochemical studies and reactivity with monoclonal antibodies directed at differentiation markers failed to show alterations in the distribution or maturation of monocytes. Peripheral blood mononuclear cells (PBMC) from these individuals produced lower levels of interleukin-1 (IL-1) in response to bacterial lipopolysaccharide and Staphylococcus albus stimulation as compared to the other S. mansoni infected individuals. Overall, IL-1 production by PBMC stimulated with LPS or Staphylococcus albus was higher in infected individuals compared to uninfected controls and correlated with intensity of infection (r = 0.41, P = 0.002 for LPS; r = 0.45, P = 0.006 for S. albus). IL-1 expression by monocytes from individuals with heavy S. mansoni infection exceeded that of subjects with light infection. The intensity-related increase in IL-1 expression did not correlate with the maturational state or the immunoregulatory properties of the monocytes.

Manipulation of the fractured nose using topical local anaesthesia.

Simple fracture of the nasal bone can be reduced under local or general anaesthesia. A combination of topical anaesthetic (EMLA cream) and cocainization of the nasal mucosa was used successfully in manipulation of fractured nasal bones in twelve patients without discomfort.