Allergic fungal sinusitis: detection of universal fungal, aspergillus and bipolaris DNA in sinus aspirate using polymerase chain reaction and evaluation of aspergillus specific IgE

Allergic fungal sinusitis [AFS] is believed to represent a hypersensitivity reaction to fungal antigens. The pathophysiology of AFS is still not clearly understood. It is believed that it is not a true fungal infection, but an allergic response to fungal organisms that have colonized the sinus mucosa and secondarily cause a hypersensitivity reaction in the host. However, some patients with AFS do not have allergy to the fungi identified in their eosinophilic mucous but may have elevated IgE levels to other fungi. The patients are usually atopic to multiple aeroallergens. Early reports noted primarily Aspergillus species in allergic mucin, but more recently, the dematiaceous fungi, which include Bipolaris, Curvularia, Alternaria and Helmenthosporium species have been identified in most AFS cases. PCR is significantly more sensitive than nasal swabs cultures in detecting the presence of fungi in nasal mucosa. In our study, 68 cases were selected and sinus aspirates were withdrawn whereas a part of the mucous was used for fungal culture and the other part was used for PCR assay for universal fungal, Aspergillus and Bipolaris DNA. Measurement of Aspergillus specific IgE in sinus aspirate and serum total IgE were done. A control group [10 cases] was included. Among the total number of AFS [68], only 42 cases gives positive fungal growth with a percentage of 61.7% while among 10 control cases, only 3 cases gives positive growth with a percentage of 30%. Regarding AFS [42 cases], Dematiaceous family was the most common as it was isolated from 30 cases [71.4%]. Bipolaris was the most common isolated species [18 cases] followed by Curvularia [11 cases] and Alternaria [1 case]. Aspergillus family was isolated from 11 cases [26.1%]. Aspergillus fumigatus was more common as it was isolated from 8 cases followed by Aspergillus niger [3 cases]. The results of PCR assay assured the detection of fungal DNA in all cases of AFS group [68 cases] and in 4 cases of control group [40%]. Aspergillus DNA was detected in 15 cases [22.05%] while Bipolaris DNA was detected in 27 cases [39.70%]. Ten patients were positive for Aspergillus fumigatus specific IgE [14.7%] out of 68 patients and the mean value was 11.32 +/- 4.12 IU/ml which was significantly higher than the mean value of this specific IgE in our control group which was 0 IU/ml. Also, only 7 patients from the above 10 patients were positive to Aspergillus fumigatus by PCR [5 only gives positive culture] and this indicates that 3 patients were negative to Aspergillus fumigatus either by culture or PCR but they showed Aspergillus fumigatus allergen specific IgE, on the other hand, 8 cases were positive to Aspergillus fumigatus by PCR and 3 cases were positive by culture failed to show any Aspergillus fumigatus specific IgE indicating that the presence of fungus is not essentially accompanied with allergic process

Bacterial DNA and its consequences in patients with cirrhosis and culture negative, non neutrocytic ascites

The detection of bacterial DNA [BactDNA] in serum and ascitic fluid [AF] from patients with liver cirrhosis and ascites is interpreted as molecular evidence of intestinal bacterial translocation [BT] and considered sufficient to activate the cellular immune response leading to greater cytokine synthesis. In the present work we study whether BactDNA and Tumor necrosis factor-alpha [TNF- alpha] in cirrhotic patients with culture negative, non neutrocytic ascites have been implicated in various complications of cirrhosis such as hepatorenal syndrome [HRS], spontaneous bacterial peritonitis [SBP] and mortality. We studied 34 patients with liver cirrhosis and culture negative, non neutrocytic ascites [22 patients without BactDNA [group I] aged [48.3 +/- 7.85y] and 12 patients with BactDNA [group II], aged [49.7 +/- 6.5y]]. Full history and complete clinical examination were done with the following investigations in the first admission and subsequent admissions during follow up for 24 weeks: complete blood picture, S. creatinine, S. bilirubin, S. albumin, S. transaminases [AIT and AST], AF and plasma TNF-alpha, AF protein and polymorphnuclear leucocytes [PMNL], both blood culture and AF aerobic and anaerobic cultivation, and detection of blood and ascitic fluid BactDNA using PCR. Plasma and ascitic TNF-alpha were significantly higher in cirrhotic patients with compared to those without BactDNA during first admission [54.5 +/- 22.56 vs 35.2 +/- 17.97; 123.2 +/- 49.32 vs 82.6 +/- 29.58 pg/ml respectively, P<0.05]. These changes became highly significant at the end of follow up of both groups [119.3 +/- 27.19 vs 40.2 +/- 16.08; 518.8 +/- 91.11 vs 97.6 +/- 17.81 pg/ml respectively, P<0.001]. There is non significant change of plasma and ascitic TNF-alpha in group I at first admission compared to those at the end of follow up [P>0.05]. However, in group II, there is highly significant increase in both plasma and ascitic TNF-alpha at the end of follow up compared to those at the first admission [P<0.001]. The relative risk of deaths, HRS and SBP were higher in patients with compared to those without BactDNA after follow up for 24 weeks [2.73, 27.37 and 18.18 respectively]. There were significant positive correlation between both plasma and ascitic TNF-alpha and each of serum creatinine and PMNL in the studied patients at the end of follow up. [r= 0.590, p= 0.002 ; r= 0.535, p= 0.005 ; r=0.499, p=0.009 ; r= 0.589, p= 0.002, respectively]. -Patients with BactDNA had more advanced liver disease after 24 weeks follow up compared to patients without BactDNA. We conclude that cirrhotic patients with culture negative, non neutrocytic ascites and BactDNA have significant higher level of AF and plasma TNF-alpha and higher risk of HRS, SBP and morality compared to those without BactDNA during follow up for 24 weeks which could suggest that both BactDNA and TNF-alpha have been implicated in these complications of liver cirrhosis

Nosocomial infections caused by klebsiellae species with plasmids carrying bla shv-5 extended-spectrum beta-lactamase gene in Mansoura university hospitals

SHV-5 is a variant of SHV-1 and it is considered now one of the most important ESBL enzymes produced by Klebsiellae species. In this study we aimed to detect the prevalence of plasmid encoded SHV-5 among Klebsiellae strains causing nosocomial infections in Mansoura University Hospitals [MUH]. One hundred and seventy four Klebsiellae strains were isolated from overall 680 cases of nosocomial infections [25.59%] acquired within MUH over 4 months period from July to November 2004. One hundred and thirty six isolates [78.16%] of them were K. pneumoniae and 38 isolates [21.84%] were K. oxytoca. MICs [micro g/ml] of the isolated strains was done for augmentin, cefoperazone and ceftazidime using E test. One hundred and forty one [81.03%] of them were beta- lactamase producer as detected by nitrocefin discs, where 77 isolates from ?-lactamase producing strains were ESBL producers constituting 44.25% of Klebsiellae isolates. Sixty six ESBL producing strains of total 77 were isolated from cases of blood stream infections [85.71%]. Sixty five ESBL producing strains were isolated from neonatal intensive care unit [NICU] [84.42%]. All ESBL producing strains [n=77] posses at least one large plasmid > 23 kbp. SHV-5 gene was amplified by PCR after plasmid isolation from ESBL producing Klebsiellae isolates, reveal that 68 isolates [88.31%] were harbored SHV-5 gene on their large plasmids. In conclusion we found that the SHV-5-producing K. pneumoniae isolates were recovered from different wards of Mansoura University Hospital during the studied period. Thus, it was hypothesized that one clone may have persisted in that hospital. We recommend that infection control measure of endemic ESBL producers should include : the consumption of the broad-spectrum cephalosporins needs to be restricted to reduce the selection pressure which enables the proliferation of ESBL producers in hospital, continuous application of infection control program as; surveillance, hand washing and contact isolation procedure

Detection of bordetella pertussis by PCR in infants with respiratory failure

Despite being under-reported, Bordetella pertussis injection remains a severe disease of high incidence world-wide. No cases were reported in Egypt since 2001. Different immunization protocols exist in different countries with variable vaccination coverage ratios. This prospective investigation study was conducted in the PICU of Mansoura University Children Hospital. Mansoura, Egypt identifying cases of B. pertussis infection among mechanically ventilated infants presenting with respiratory failure and features compatible with pertussis [bronchopneumonia, apnoea, acute life threatening event]. Infants less than one year of age were enrolled over a period of 12 months. Sixty one specimens of endotracheal secretions were examined by PCR for the presence of a 262-bp target sequence from IS481 specific for B. pertussis. Nine specimens were positive for B. pertussis, five infants in this group did not survive. All non survivors were younger than 6 months of age. Infants in the PCR-positive group had a younger age [p = 0.038], a longer duration of illness prior to PICU admission [p < 0.01] and a higher mortality rate [p = 0.045] compared to the PCR - negative infants. It is crucial to raise awareness, among medical professionals, of clinical picture, complications and treatment of pertussis, If immunization program of Egypt was to be reviewed, there may be a need for a more accelerated primary immunization program against pertussis with booster doses for young adults

Renal impairment and mortality in cirrhotic patients with spontaneous bacterial peritonitis: role of proinflammatory cytokines and a possible role of intercellular adhesion molecule-1

This study included 32 cirrhotic patients with spontaneous bacterial peritonitis [SBP] aged 36.3 +/- 5.3 years, 15 cirrhotic patients with sterile ascitic fluid aged 37.1 +/- 4.1 years and 15 age- matched healthy control subjects. Serum bilirubin, serum albumin, serum creatinine, CBC, ascitic fluid polymorphonuclear leucocytes [PMNL], plasma and ascitic fluid levels of TNF-alpha, IL-6 and sICAM-1, abdominal X-ray and ultrasonography were assessed. It was concluded that cirrhotic patients with SBP have an abnormal inflammatory and immune responses in terms of increased ascitic fluid and plasma levels of TNF-alpha, IL-6 and sICAM-1. SICAM-1 implies a favorable condition for PMNL migration towards the peritoneum accentuating the inflammatory and immune responses which may be an important mechanism of SBP that induced renal impairment and death