Real-time PCR versus MALDI-TOF MS and culture-based techniques for diagnosis of bloodstream and pyogenic infections in humans and animals.

AIMS: This study was applied to evaluate the usefulness of a high-throughput sample preparation protocol prior to the application of quantitative real-time PCR (qPCR) for the early diagnosis of bloodstream and pyogenic infections in humans and animals compared to matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and classical culture. METHODS AND RESULTS: Saponin-mediated selective host cell lysis combined with DNase-1 was applied for processing of whole blood and pus clinical samples collected from suspected cases of septicaemia and pyogenic infections in humans and animals. The pre-PCR processing strategy enabled the recovery of microbial cells with no changes in their colony forming units immediately after the addition of saponin. DNase-1 was efficient for removing the DNAs from the host cells as well as dead cells with damaged cell membranes. The metagenomic qPCR and MALDI-TOF MS could identify the bacterial community of sepsis at species level with a concordance of 97·37% unlike the conventional culture. According to qPCR results, Staphylococcus aureus (24·24%) was predominated in animal pyogenic infections, whereas Klebsiella pneumonia (31·81%) was commonly detected in neonatal sepsis. CONCLUSIONS: Saponin combined with DNase-1 allowed the efficient recovery of microbial DNA from blood and pus samples in sepsis using qPCR assay. SIGNIFICANCE AND IMPACT OF THE STUDY: Metagenomic qPCR could identify a broad range of bacteria directly from blood and pus with more sensitivity, higher discriminatory power and shorter turnaround time than those using MALDI-TOF MS and conventional culture. This might allow a timely administration of a prompt treatment.

[The additional criterion for the diagnosis of purulent-destructive forms of acute pyelonephritis].

AIM: to develop an additional criterion for objective diagnosis of purulent-destructive forms of acute pyelonephritis. MATERIALS AND METHODS: A total of 35 blood samples from patients with acute pyelonephritis aged from 19 to 85 years (mean age was 52 years) were studied. For the analysis, a classical bacteriological method of isolating blood culture and a modern molecular genetic method of fluorescent in situ hybridization (FISH) using DNA probes complementary to species-specific regions of the 16S rRNA gene of microorganisms were used. The obtained results were processed using the Statistica 6.0 program. RESULTS: The bacteriological study showed that only in one patient (2.9%) a blood culture was positive, whereas with FISH, polymicrobial infection consisting of two associates was observed in five patients (14.3%), and 207 specific luminescence from different DNA probes were detected in blood samples. The associations of members of the Enterobacteriaceae with S. aureus were 2.4 times more frequent than associations of the Enterobacteriaceae with S. epidermidis, indicating a development of the polymicrobial infection and, accordingly, the development of purulent stage of acute pyelonephritis. CONCLUSION: For the diagnosis of acute pyelonephritis, we suggest to use an additional criterion, which consists in applying of FISH method for the detection and simultaneous identification of bacteria that were adhered on the surface of erythrocytes and located intracellularly. This method may allow for the differential diagnosis of serous and purulent stages of acute pyelonephritis.

[Diagnosis and treatment of suppurative mediastinitis - a special chapter in the history of surgery (in Russian only)]. / Diagnostika i lechenie gnoinogo mediastinita - osobaia glava v istorii khirurgii.

It was reviewed a history of diagnosis and treatment of suppurative mediastinitis from ancient times to our time depending on inflammation type, localization and clinical features. An important role of national surgical school in the development of surgical treatment was emphasized.

Suppurative Cervical Tuberculous Lymphadenitis Mimicking a Metastatic Neck Mass.

Tuberculosis is an important public's health problem in developing countries. Although tuberculosis is commonly found in lungs, it could also be found in lymph nodes, mouth, tonsils, tounge, nose, epiglottis, larynx, and pharynx of head-neck region. The most common form of extrapulmonary tuberculosis placed outside the lungs is tuberculous lymphadenitis. Tuberculosis lymphadenitis is the most commonly seen form of the extrapulmoner tuberculosis and usually invades the lymph nodules of the cervical region. An adult patient with tuberculous lymphadenitis has been presented within the context of the literature reviewed. A 42-year-old woman, who has admitted to our clinic with a fistulized mass in the right side of her neck, has been diagnosed and treatment has been performed. No other problems were found in the systemic examination of the patient. Since there is no specific finding of pulmonary tuberculosis, detailed investigations are required in patients admitted with the sypmtomps of neck mass.

[Pustular psoriasis]. / Pustulöse Psoriasis.

A number of pustular skin diseases share clinical, pathogenetic, and epidemiological aspects with plaque-type psoriasis, and their classification as a separate clinical entity or as a subtype of psoriasis remains controversial, which is also reflected in the multitude of their names. They include generalized pustular psoriasis with its subtypes, acrodermatitis continua suppurativa (Hallopeau), acute pustulosis palmopantaris, palmoplantar pustular psoriasis, and pustular variants of a mostly TNF-blocker triggered paradoxical psoriasiform dermatitis. In this article, the epidemiology, clinical picture, pathogenesis, genetics, and therapy of these pustular skin diseases are described.

[HLA-DR expression on monocytes as a prognostic criterion for the development of pyoinflammatory processes at various stages of surgical treatment].

We studied relative content of CD14+HLA-DR in 214 patients including those with aseptic instability and stable implants of the knee and the hip, periprosthetic infection of the hip, chronic posttraumatic osteomyelitis of long tubular bones, and osteomyelitis generalization (sepsis). We determined HLA-DR expression on monocytes by laser flow cytofluorometry using Beckman Coulter Epics XL (USA) cytometer and monoclonal antibodies (Immunotech Company, France). It has been demonstrated, that the decrease in HLA-DR expression on monocytes can be one of the mechanisms of increasing immune disorders in patients with orthopedic pathology and monitoring HLA-DR monocytic expression can be used as a prognostic criterion of developing pyoinflammatory processes at the stages of surgical treatment.

[The value of volatile fatty acids, citrulline and malondialdehyde for diagnosis of suppurative cholangitis in obstructive jaundice and choosing of optimal surgical approach (with commentary)]. / Otsenka informativnosti pokazatelei letuchikh zhirnykh kislot, tsitrullina i malonovogo dial'degida dlya diagnostiki gnoinogo kholangita pri mekhanicheskoi zheltukhe i vybora optimal'noi khirurgicheskoi taktiki (s kommentariem).

AIM: To define the role of volatile fatty acids, citrulline and malondialdehyde for diagnosis of suppurative cholangitis in obstructive jaundice and to define optimal surgical approach. MATERIAL AND METHODS: We studied the results of examination and treatment of 87 patients with different hepatopancreatobiliary pathology complicated by obstructive jaundice. It was determined blood concentration and range of volatile fatty acids which are metabolites of facultative anaerobic and obligate anaerobic bacterial pathogens. RESULTS: In 39 patients the defined levels of acetic, propionic, butyric and isovaleric acids contributed to accurate diagnosis of suppurative cholangitis. Statistically significant threshold concentrations of volatile fatty acids were revealed. These values facilitate detection of anaerobic microflora in suppurative cholangitis. CONCLUSION: Research of contents of volatile fatty acids, citrulline and malondialdehyde contributes to early diagnosis of suppurative cholangitis in patients with obstructive jaundice and further objectifies an algorithm of surgical tactics.

[DETERMINATION OF A FATTY ACIDS CONTENT IN BILIARY LIPIDS AS A PROGNOSTIC FACTOR FOR THE COMPLICATIONS OCCURRENCE IN PATIENTS AFTER OPERATIONS FOR AN ACUTE CHOLECYSTITIS IN A HIGH OPERATIVE­ANESTHESIOLOGICAL RISK].

Investigation was conducted with objective to establish a prognostic significance of the fatty acids content in biliary lipids for the complications occurrence in patients, operated on for an acute cholecystitis, while presence of high operative­anesthesiological risk. A fatty acids content in biliary lipids was analyzed in 64 patients in presence of moderate (grade ІІ) and severe (grade ІІІ) course of the disease. Coefficients К1 and К2 for prognostication of postoperative purulent­inflammatory complications occurrence were elaborated, their threshold level (К1=0.59, К2=1.8) was established. There was established, that excess of the threshold meaning of К1 and К2 in a bile of patients, suffering an acute cholecystitis while presence of high operative­anesthesiological risk, constitutes a prognostic criterion for the postoperative purulent­inflammatory complications occurrence

[PECULIARITIES OF DIAGNOSIS AND SURGICAL TREATMENT OF PATIENTS, SUFFERING ATYPICAL FORMS OF AN ACUTE APPENDICITIS].

Results of diagnosis and surgical treatment of 852 patients, suffering an acute appendicitis (AA) with atypical course, were analyzed. Retrocecal localization of appendix was noted in 61.2% of observations, a pelvic one ­ in 24.3%, medial ­ in 11.2%, and subhepatic ­ in 3.4%. Destructive forms of atypical AA were diagnosed in 92.5% patients, and various kinds of peritonitis ­ in 77.7%. Some diagnostic (rectal thermometry, test with ethanol) and operative methods (including laparoscopic) in destructive forms of AA, complicated by typhlitis, were improved and tested. Diagnostic­treatment algorithm, permitting to optimize tactic of treatment and to reduce the early postoperative complications rate from 9.9 tо 3.5% (р<0.001), was proposed.

Neonatal suppurative parotitis over the last 4 decades: report of three new cases and review.

BACKGROUND: Neonatal suppurative parotitis is a rare disease. Only 32 cases were reported in the English-language literature between 1970 and 2004. METHODS: We searched Medline for acute, neonatal, bacterial, suppurative, parotitis, facial, preauricular swelling starting from 1970, limiting our search to the English-language literature. We reviewed all the reported cases together with three more managed in our department. RESULTS: We identified nine new cases since 2004. The total number of patients reviewed was 44, including our patients. Most of them were male (77%). The majority developed unilateral inflamed parotid swelling (77%) and exuded pus from the ipsilateral Stensen duct. Fever was seen in fewer than half of them (47%). Premature babies constituted a third of the patients. Staphylococcus aureus was the leading causative agent (61%). Most patients responded well to conservative treatment with antibiotics (77%). The most frequently used combination of antibiotics was an anti-staphylococcal agent with either an aminoglycoside or a third-generation cephalosporin. A minority required surgical drainage. No deaths were reported in the group studied after 1970. CONCLUSION: Neonatal suppurative parotitis is rare but easy to diagnose and if readily treated with appropriate antibiotics the outcome is excellent.

[The improvement of complex treatment of odontogenous periostitis in elderly patients].

The paper presents the results of clinical examination of 114 patients aged 60-88 years with acute odontogenous periostitis receiving treatment in in-patient maxillofacial surgery unit. The dynamic of clinical symptoms is used to carry out the comparative effectiveness study of several peroral antibiotics in elderly patients.

[Indices of tissue necrosis markers in acute pyo-destructive diseases of organs of the abdominal cavity].

Investigation of indices of lactate dehydrogenase (LDG) and creatine phosphokinase (CPK) in a group of patients with extensive necrotic alterations in organs of the abdominal cavity and in a group of patients without extensive necroses has shown that in the first group there were higher levels of LDG and CPK. It shows that the LDG and CPK indices may be used as markers of tissue necrosis of the abdominal organs. In addition, in patients who died against the background of growing polyorganic insufficiency the LDG and CPK level by the end of treatment was statistically reliably higher than on admission of the same patients that may be an indicator of growing phenomena of dystrophy and micronecroses in vitals, so the indices of LDG and CPK may show the degree of severity of polyorganic insufficiency and determine its prognosis.

[Significance of soluble DLL1 in diagnosis of intracranial infectious diseases in children].

OBJECTIVE: To investigate the significance of soluble DLL1 (Delta-like-1) levels of cerebrospinal fluid (CSF) and serum in the diagnosis of intracranial infection in children. METHODS: Fifty children with intracranial infection, including 20 cases of tuberculous meningitis (TM), 20 cases of viral meningitis (VM) and 10 cases of purulent meningitis (PM), and 20 children without intracranial infection (control group) were enrolled. The levels of soluble DLL1 in CSF and serum were measured using ELISA. RESULTS: The level of CSF soluble DLL1 in the TM group was significantly higher than that in the VM, PM and control groups (2.89 ± 1.72 ng/mL vs 0.14 ± 0.14 ng/mL, 0.27 ± 0.21 ng/mL, 0.13 ± 0.12 ng/mL; P<0.01). The level of serum soluble DLL1 in the TM group was also significantly higher than that in the VM, PM and control groups (12.61 ± 6.45 ng/mL vs 2.28 ± 2.27 ng/mL, 2.38 ± 1.79 ng/mL, 2.26 ± 2.10 ng/mL; P<0.01). The levels of soluble DLL1 in the CSF and serum in the VM and PM groups were not significantly different from those in the control group. CONCLUSIONS: Soluble DLL1 as a novel indicator might have potentially important value in the diagnosis of TM.

[Retropharyngeal abscess in an adult patient complicated by purulent meningoencephalitis].

The authors report a case of successful treatment of an adult woman presenting with retropharyngeal abscess complicated by purulent meningoencephalitis. Peculiarities of diagnostics and treatment of retropharyngeal abscess in adult patients are discussed.

Multidisciplinary Diagnostic Approach in Intraocular Lymphoma Featuring Pseudo-hypopyon: Case Series and Literature Review.

PURPOSE: Diagnosis of intraocular lymphoma (IOL) is usually achieved by histopathological analysis. However, it may lead to inconclusive results due to the scarcity of malignant cells obtained by biopsy, hence leading to delayed diagnosis. We report two cases of IOL with pseudo-hypopyon, a rare feature of IOL, as their initial ocular feature, diagnosed using a multidisciplinary diagnostic approach. Common clinical features of IOL with pseudo-hypopyon were also investigated. METHODS: Retrospective case series and literature review. RESULTS: Two cases of IOL, a 78-year-old female and a 59-year-old male, whom had been diagnosed with systemic B-cell lymphoma developed pseudo-hypopyon and visual impairment during the course of their chemotherapy. Diagnosis of IOL was achieved from anterior chamber aspiration samples with supplementary diagnostic tools including flow cytometric immunophenotyping, interleukin and IgH gene rearrangement analysis in addition to the conventional histopathological analysis. Generally, pseudo-hypopyon was more commonly seen in secondary IOL and may associate with hyphema and high intraocular pressure. CONCLUSION: Pseudo-hypopyon is a rare feature of IOL, more commonly seen in secondary IOL, which can be accompanied by hyphema and high intraocular pressure. Supplementary diagnostic tools such as flow cytometric immunophenotyping, interleukin analysis, and immunogloblin H gene rearrangement analysis are useful for supporting the diagnosis of IOL with pseudo-hypopyon.

Primary Ciliary Body Marginal Zone Lymphoma Presenting as Hemorrhagic Hypopyon.

PURPOSE: Our purpose is to report a patient with primary unilateral ciliary body marginal zone lymphoma who initially presented with hemorrhagic hypopyon. METHODS: Retrospective review of the clinical, imaging, and immunohistopathological features of the case was performed. RESULTS: A 59-year-old man was referred with right anterior uveitis of unknown etiology which was unresponsive to systemic treatment. Slit-lamp biomicroscopy showed normotensive hemorrhagic hypopyon in that eye. Anterior segment ultrasound biomicroscopy revealed an iridociliary mass lesion. Because an anterior chamber paracentesis was noncontributory, a diagnostic cyclectomy was performed. Histopathological evaluation showed that the neoplastic cells were positive for CD20, lambda light chain, and BCL 2. BCL 6, CD10, CD5, SOX11, kappa, and Cyclin D1 stains were negative. The final diagnosis was extranodal marginal zone lymphoma of the ciliary body. CONCLUSIONS: Although rare, ciliary lymphoma may be a cause of intractable anterior uveitis. Repeat biopsies could be carried out when there is a high level of clinical suspicion.

Hypopyon in patients with uveitis.

PURPOSE: To evaluate the risk of and risk factors for hypopyon among patients with uveitis and to evaluate the risk of visual changes and complications after hypopyon. DESIGN: Retrospective cohort study. PARTICIPANTS: Patients with uveitis at 4 academic ocular inflammation subspecialty practices. METHODS: Data were ascertained by standardized chart review. MAIN OUTCOME MEASURES: Prevalence and incidence of hypopyon, risk factors for hypopyon, and incidence of visual acuity changes and ocular complications after hypopyon. RESULTS: Among 4911 patients with uveitis, 41 (8.3/1000) cases of hypopyon were identified at the time of cohort entry. Of these, 2885 initially free of hypopyon were followed over 9451 person-years, during which 81 patients (2.8%) developed hypopyon (8.57/1000 person-years). Risk factors for incident hypopyon included Behçet's disease (adjusted relative risk [RR]=5.30; 95% confidence interval [CI], 2.76-10.2), spondyloarthropathy (adjusted RR=2.86; 95% CI, 1.48-5.52), and human leukocyte antigen (HLA)-B27 positivity (adjusted RR=2.04; 95% CI, 1.17-3.56). Patients with both a spondyloarthropathy and HLA-B27 had a higher risk than either factor alone (crude RR=4.39; 95% CI, 2.26-8.51). Diagnosis of intermediate uveitis (+/- anterior uveitis) was associated with a lower risk of hypopyon (with respect to anterior uveitis only, adjusted RR=0.35; 95% CI, 0.15-0.85). Hypopyon incidence tended to be lower among patients with sarcoidosis (crude RR=0.22; 95% CI, 0.06-0.90; adjusted RR=-0.28; 95% CI, 0.07-1.15). Post-hypopyon eyes and eyes not developing hypopyon had a similar incidence of band keratopathy, posterior synechiae, ocular hypertension, hypotony, macular edema, epiretinal membrane, cataract surgery, or glaucoma surgery. Post-hypopyon eyes were more likely than eyes not developing hypopyon to gain 3 lines of vision (crude RR=1.54; 95% CI, 1.05-2.24) and were less likely to develop 20/200 or worse visual acuity (crude RR=0.41; 95% CI, 0.17-0.99); otherwise, visual outcomes were similar in these groups. CONCLUSIONS: Hypopyon is an uncommon occurrence in patients with uveitis. Risk factors included Behçet's disease, HLA-B27 positivity, and spondyloarthropathy. Intermediate uveitis cases (+/- anterior uveitis) had a lower risk of hypopyon. On average, post-hypopyon eyes were no more likely than other eyes with uveitis to develop structural ocular complications or lose visual acuity.