Investigation of Pneumocystis jirovecii in Lung Cancer Patients with the Nested PCR Method

Objective: Pneumocystis jirovecii (P. jirovecii) is an opportunistic pathogen in humans. Early diagnosis and optimal treatment of patients with P. jirovecii pneumonia (PJP) remains a key priority. This study investigated P. jirovecii in patients with lung cancer using the nested-polymerase chain reaction (PCR) method and examined the relationship between P. jirovecii and clinical findings. Methods: The study included 60 patients with lung cancer and 30 patients without lung cancer. The bronchoalveolar lavage (BAL) fluid samples of these 90 individuals were taken for diagnostic purposes in the University of Health Sciences Turkey, Van Training and Research Hospital, Clinic of Chest Diseases. Patient information was recorded. After DNA isolation from the BAL fluid samples taken from patients, the nested-PCR protocol for amplification of mtLSUrRNA in P. jirovecii was performed. Results: P. jirovecii DNA was detected in 40 (66.67%) of the lung cancer patients included in the study and in six (20%) patients without lung cancer, that is, in 46 (51.11%) patients. The rate of nested-PCR positivity in the lung cancer group was significantly higher than that in the non-lung cancer group (p=0.0001). Additionally, a statistically significant correlation was found between anorexia and weight loss, fever and sputum P. jirovecii positivity in patients with lung cancer (p<0.005). Conclusion: These findings suggest that lung cancer patients should be evaluated for PJP.

Genetic Diversity of Echinococcus granulosus Isolated from Humans: A Comparative Study in Two Cystic Echinococcosis Endemic Areas, Turkey and Iran.

Cystic echinococcosis (CE) is one of the most important zoonotic parasitic diseases caused by the larval stage of Echinococcus granulosus. Based on molecular studies and DNA sequencing, E. granulosus has been classified into 10 different genotypes (G1 to G10). Two neighboring countries, Turkey and Iran, are considered the two main foci of CE in the Middle East. The current study is aimed at examining the genotype diversity of E. granulosus isolated from human clinical samples in Turkey and Iran. Surgically removed human hydatid cysts were collected from East Azerbaijan and Fars provinces in Iran and Van province in Turkey. After extracting DNA, performing PCR, targeting the cox1 gene, the PCR products were purified from the gel and were sequenced from both directions. The sequences were aligned and compared, using BioEdit and also the BLAST program of GenBank. The maximum likelihood tree was constructed based on the Tamura-Nei model, using the MEGAX software. Phylogenetic analysis showed that the human isolated samples were classified into two major clades: G1 (from Iran and Turkey) and G3 (5 samples from northwestern Iran and one sample from Turkey). The mean and degree of genetic divergence (K2P) between the two major clades, G1 and G3, were 0.2% and 0.7 ± 0.4%, respectively. The findings of the current study revealed that the sheep strain (G1) and the less important strain G3 have major roles in the transmission cycle of CE in two neighboring countries, Iran and Turkey. Therefore, it is necessary to interpose the life cycle of this parasite and reduce the disease burden in livestock and humans by adopting common regional preventive and control policies.

Molecular characterization of human lung and liver cystic echinococcosis isolates in Van Province, Turkey.

Cystic echinococcosis (CE) is a zoonotic infection and could lead to significant public health problems. The genetic diversity of CE includes five species: E. granulosus sensu stricto (s.s.) (G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5), Echinococcus canadensis genotypic cluster (G6, G7, G8 and G10, with the doubtful G9) and the Echinococcus felidis (lion strain). The species are important in epidemiology, pathology, control, prevention measures and vaccine/drug designs. The aim of the present study was to determine the E. granulosus genotypes in humans in the Van province in east of Turkey. In total, 102 echinococcal cysts were collected from operated patients. Genomic analyses were conducted with PCR-RFLP of the rDNA internal transcribed spacer 1 (ITS1) fragment and partial PCR sequencing of the cytochrome c oxidase subunit 1 (cox1) mitochondrial DNA gene region. In total, DNAs of 96 isolates could be extracted, unfortunately six extractions failed. The PCR-RFLP analysis findings were identical in all isolates. Two bands were observed at approximately 300 bp and 600 bp. All profiles corresponded to the G1-G3 strain. Also, 446 bp amplified gene regions were observed for cox1. Out of 20 samples, alignment of 16 sequences exhibited a total identification (100%) of granulosus sensu stricto (G1/G3). Of 16 samples, 8 were obtained in the lung and 12 were obtained in the liver; 8 belonged to male and 12 belonged to female patients. Other four samples exhibited one nucleotide substitution at different positions. Four samples had one nucleotide substitution at different positions. We detected single nucleotide variations in TRH1, TRH67, TRH85 and TRH89 isolates at the positions C240T; G330T; G211A and T157C, respectively. In conclusion, the present study was the first comprehensive molecular investigation on genetic characterization of human CE isolates in Van region. The findings demonstrated that E. granulosus s.s. was the dominant species, which indicated that the sheep-dog cycle was the source in human infections. And, probably, it would be possible to describe these mutations as "Turkey" or "lung" variants. In addition to contributing molecular epidemiological data, the present results should be considered when designing and implementing E. granulosus control programs.

A Rare Case of Secondary Hydatid Cyst: Uterus and Colon Locations in the Same Patient

The aim of the present study was to present a case with secondary hydatid cysts in both uterus and colon. The patient was a 71-year-old female living in Hakkari, Turkey. She was admitted to the Van Yuzuncu Yil University Faculty of Medicine Medical Center with complaints of chronic abdominal and pelvic pain, and swelling in the abdomen. First, the sagittal T2 weighted magnetic rezonance imaging (MR) showed a type-3 cyst hydatid with daughter vesicles located at the posterior of uterus. Later, MR revealed a type-2 cystic lesion with detached membrane adhered to the anterior wall of colon and it was reported to be associated with abdomen. When the previous liver surgery history of the patient was kept in mind, the new finding was suggestive of a secondary cystic hydatid . In conclusion, it is possible to diagnose secondary cystic echinococcosis in patients with a history of primary cyst surgery in liver or any other organ by combining the symptoms and imaging findings.