OBJECTIVE: To conduct eukaryotic expression of the leucine-rich repeat containing 15 (LRRC15), a differentially expressed protein in excretory secretory antigens of Taenia solium cysticercus, and predict its antigen epitope. METHODS: The molecular weight, stability, amino acid sequence composition, isoelectric point and T lymphocyte epitope of the LRRC15 protein were predicted using the bioinformatics online softwares ExPASy-PortParam and Protean. The full-length splicing primers were designed using PCR-based accurate synthesis, and the LRRC15 gene was synthesized. The recombinant pcDNA3.4-LRRC15 plasmid was constructed and transfected into HEK293 cells to express the LRRC15 protein. In addition, the LRRC15 protein was characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. RESULTS: The recombinant pcDNA3.4-LRRC15 plasmid was successfully constructed, which expressed the target LRRC15 protein with an approximately molecular weight of 70 kDa. Bioinformatics prediction with the ExPASy-PortParam software showed that LRRC15 was a hydrophilic protein, which was consisted of 644 amino acids and had a molecular weight of 69.89 kDa and an isoelectric point of 5.6. The molecular formula of the LRRC15 protein was C₃₀₇₃H₄₉₄₂N₈₄₆O₉₅₃S₂₈ and had an instability coefficient is 50.3, indicating that LRRC15 was an instable protein. Bioinformatics prediction with the Protean software showed that the dominant T-cell antigen epitopes were located in 292 to 295, 353 to 361, 521 to 526 and 555 to 564 amino acids of the LRRC15 protein, and the T-cell antigen epitopes with a high hydrophilicity, good flexibility, high surface accessibility and high antigenicity index were found in 122 to 131, 216 to 233, 249 to 254, 333 to 343, 358 to 361, 368 to 372, 384 to 386, 407 to 412, 445 to 450, 469 to 481, 553 to 564, 588 to 594, 607 to 617 and 624 to 639 amino acids. Following transfection of the recombinant pcDNA3.4-LRRC15 plasmid into HEK293 cells, SDS-PAGE and Western blotting identified LRRC15 proteins in cell secretory culture media, cell lysis supernatants and sediments. The LRRC15-His fusion protein was purified from the cell culture medium, and SDS-PAGE identified a remarkable band at approximately 70 kDa, while Western blotting successfully recognized the band of the recombinant LRRC15 protein. CONCLUSIONS The eukaryotic expression and antigen epitope prediction of the LRRC15 protein in the excretory secretory antigens of T. solium cysticercus have been successfully performed, which provides insights into further understandings of its biological functions.
Amino Acids ; Animals ; Antigens, Helminth/genetics* ; Cysticercus/genetics* ; Epitopes/genetics* ; Eukaryota ; HEK293 Cells ; Humans ; Leucine-Rich Repeat Proteins ; Membrane Proteins ; Taenia solium/genetics*

The prevalence of human taeniasis has decreased in Korea. The stool egg positive proportion decreased from 1.9% in 1971 to 0% in 2004 in nationwide surveys. The neurocysticercosis (NCC) is also presumed to decrease. However, detailed information regarding the recent status of NCC in Korea is lacking. We retrospectively reviewed NCC cases from 1990 to 2016 at Asan Medical Center, a 2700-bed tertiary referral hospital in Korea. We identified patients based on clinical symptoms, brain imaging, pathology and serological assay. The cases were classified as parenchymal, extraparenchymal, and mixed NCC. Eighty-one patients were included in the analysis. The mean age was 54.5 years, and 79.0% were male. The number of NCC cases was highest from 1995 to 1999, and continuously decreased thereafter. Forty (49.4%) patients had parenchymal NCC, while 25 (30.9%) patients had extraparenchymal NCC, and 16 (19.8%) patients had mixed NCC. The seizure and headache were most common symptom of parenchymal NCC and extraparenchymal NCC respectively. Hydrocephalus was more common in extraparenchymal NCC, and patients with extraparenchymal NCC were more likely to require a ventriculoperitoneal shunt. Cases of NCC are decreasing accordingly with human taeniasis and lesion location was the most important determinant of clinical presentation and outcome of NCC in Korea.
Chungcheongnam-do ; Headache ; Humans ; Hydrocephalus ; Korea ; Male ; Neurocysticercosis ; Neuroimaging ; Ovum ; Pathology ; Prevalence ; Retrospective Studies ; Seizures ; Taenia solium ; Taeniasis ; Tertiary Care Centers ; Ventriculoperitoneal Shunt

Cysticercosis, a parasitic disease caused by Taenia solium metacestode (TsM), has a major global public health impact in terms of disability-adjusted life years. The parasite preferentially infects subcutaneous tissue, but may invade the central nervous system, resulting in neurocysticercosis (NC). NC is an important neglected tropical disease and an emerging disease in industrialized countries due to immigration from endemic areas. The prevalence of taeniasis in Korea declined from 0.3%–12.7% during the 1970s to below 0.02% since the 2000s. A survey conducted from 1993 to 2006 revealed that the percentage of tested samples with high levels of specific anti-TsM antibody declined from 8.3% to 2.2%, suggesting the continuing occurrence of NC in Korea. Modern imaging modalities have substantially improved the diagnostic accuracy of NC, and recent advances in the molecular biochemical characterization of the TsM cyst fluid proteome also significantly strengthened NC serodiagnosis. Two glycoproteins of 150 and 120 kDa that induce strong antibody responses against sera from patients with active-stage NC have been elucidated. The 150 kDa protein showed hydrophobic-ligand binding activities and might be critically involved in the acquisition of host-derived lipid molecules. Fasciclin and endophilin B1, both of which play roles in the homeostatic functions of TsM, showed fairly high antibody responses against calcified NC cases. NC is now controllable and manageable. Further studies should focus on controlling late-onset intractable seizures and serological diagnosis of NC patients infected with few worms. This article briefly overviews diagnostic approaches and discusses current issues relating to NC serodiagnosis.
Antibody Formation ; Central Nervous System ; Cyst Fluid ; Cysticercosis ; Developed Countries ; Diagnosis ; Emigration and Immigration ; Glycoproteins ; Humans ; Immunologic Tests ; Korea ; Neurocysticercosis ; Parasites ; Parasitic Diseases ; Prevalence ; Proteome ; Public Health ; Republic of Korea ; Seizures ; Serologic Tests ; Subcutaneous Tissue ; Taenia solium ; Taenia ; Taeniasis

Neurocysticercosis (NCC) by Taenia solium is the most common parasitic infection of the central nervous system involving the cerebrum. However, spinal involvement of NCC is rare. Spinal NCC can cause radiculopathy, myelopathy, cauda equina syndrome, and even paraparesis, depending on its location and size. Spinal NCC may require surgical treatment as a first-line treatment because medical therapy can further aggravate the inflammation due to dead cysts, resulting in clinical deterioration. The current standard therapy for spinal NCC is surgical decompression followed by medical therapy. We experienced a case of widespread thoracolumbar intradural extramedullary cysticercosis involving the spinal canal with cerebral cysticercosis. We report this rare case with literature review.
Central Nervous System ; Cerebrum ; Cysticercosis* ; Decompression, Surgical ; Humans ; Inflammation ; Neurocysticercosis ; Paraparesis ; Polyradiculopathy ; Radiculopathy ; Spinal Canal* ; Spinal Cord Diseases ; Spine ; Taenia solium

Previous paleoparasitological studies of Joseon specimens established that the prevalence of Taenia infection was not much different from that of the early 20th century Korean population. As many of taeniases originally diagnosed as Taenia saginata in South Korea were revealed to be actually Taenia asiatica, which share a common intermediate host with T. solium (the pig), Joseon people must have ingested raw pork frequently. However, the current examination of extant Joseon documents revealed that the population ate significant amounts of beef even if the beef ban was enforced; and pork was not consumed as much as we thought. Considering the meat consumption pattern at that time, Joseon people should have been infected by T. saginata more frequently than T. asiatica. This may suggest a low prevalence of T. saginata metacestodes in cattle compared to that of T. asiatica metacestodes in pigs, possibly due to the traditional way of rearing pigs (using human feces). This letter gives us a chance to reconsider the existing preconception about parasitic infections in Korean history though we are still hard to accurately estimate the historical patterns of taeniases at this stage.
Animals ; Cattle ; Humans ; Korea* ; Meat* ; Population ; Prevalence ; Red Meat ; Swine ; Taenia saginata ; Taenia solium ; Taenia* ; Taeniasis*

Neurocysticercosis is an infection of the central nervous system caused by the larval form of the pork tapeworm Taenia solium. In the brain it occurs in two forms: parenchymal and extraparenchymal or racemose cysts. The clinical presentation of racemose cysts is pleomorphic, and is quite different from parenchymal cysticercosis. The clinical diagnosis of racemose cysts is quite challenging, with neuroimaging being the mainstay. However, the advent of newer brain imaging modalities has made a more accurate diagnosis possible. The primary focus of this article is racemose neurocysticercosis and its multitude manifestations, and includes a discussion of the newer diagnostic modalities and treatment options.
Brain ; Central Nervous System ; Cysticercosis ; Diagnosis ; Neurocysticercosis* ; Neuroimaging ; Taenia solium

The tapeworm Taenia solium is an important human zoonotic parasite that causes great economic loss and also endangers public health. At present, an effective vaccine that will prevent infection and chemotherapy without any side effect remains to be developed. In this study, codon usage patterns in the T. solium genome were examined through 8,484 protein-coding genes. Neutrality analysis showed that T. solium had a narrow GC distribution, and a significant correlation was observed between GC12 and GC3. Examination of an NC (ENC vs GC3s)-plot showed a few genes on or close to the expected curve, but the majority of points with low-ENC (the effective number of codons) values were detected below the expected curve, suggesting that mutational bias plays a major role in shaping codon usage. The Parity Rule 2 plot (PR2) analysis showed that GC and AT were not used proportionally. We also identified 26 optimal codons in the T. solium genome, all of which ended with either a G or C residue. These optimal codons in the T. solium genome are likely consistent with tRNAs that are highly expressed in the cell, suggesting that mutational and translational selection forces are probably driving factors of codon usage bias in the T. solium genome.
Animals ; Base Sequence ; Codon/*genetics ; Evolution, Molecular ; *Genome, Helminth ; Helminth Proteins/*genetics ; Molecular Sequence Data ; Taenia solium/*genetics

BACKGROUND: Cysticercosis is the most frequent parasitic infection of the nervous system. Most lesions are intracranial, and spinal involvement is rare. We describe here in two cases of neurocysticercosis (NCC) in the brain and one in the spinal cord that illustrate three distinct mechanisms leading to symptomatic acute hydrocephalus. CASE REPORT: Hydrocephalus was related to intracranial NCC in two of them. In the first case the hydrocephalus was due to an extensive arachnoiditis to the craniocervical junction, while in the second it was caused by obstruction of Magendie's foramen in the fourth ventricle by the scolex of Taenia solium. For the third patient, hydrocephalus revealed cysticercosis of the cauda equina due to the scolex. CONCLUSIONS: NCC should be considered as a possible diagnosis for patients suffering from hydrocephalus when they originate from or have traveled in endemic areas, MRI of the spine is mandatory to search for intraspinal lesions.
Arachnoid ; Arachnoiditis ; Brain ; Cauda Equina ; Cysticercosis ; Diagnosis ; Fourth Ventricle ; Humans ; Hydrocephalus* ; Magnetic Resonance Imaging ; Nervous System ; Neurocysticercosis* ; Spinal Cord ; Spine ; Taenia solium

Wnt proteins are a family of secreted glycoproteins that are evolutionarily conserved and considered to be involved in extensive developmental processes in metazoan organisms. The characterization of wnt genes may improve understanding the parasite's development. In the present study, a wnt4 gene encoding 491amino acids was amplified from cDNA of metacestodes of Taenia solium using reverse transcription PCR (RT-PCR). Bioinformatics tools were used for sequence analysis. The conserved domain of the wnt gene family was predicted. The expression profile of Wnt4 was investigated using real-time PCR. Wnt4 expression was found to be dramatically increased in scolex evaginated cysticerci when compared to invaginated cysticerci. In situ hybridization showed that wnt4 gene was distributed in the posterior end of the worm along the primary body axis in evaginated cysticerci. These findings indicated that wnt4 may take part in the process of cysticerci evagination and play a role in scolex/bladder development of cysticerci of T. solium.
Animals ; Base Sequence ; Cysticercosis/pathology ; Cysticercus/enzymology/*genetics ; DNA, Helminth/*genetics ; Gene Expression Regulation ; Humans ; In Situ Hybridization ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sus scrofa ; Swine ; Swine Diseases ; Taenia solium/embryology/enzymology/*genetics ; Wnt4 Protein/*genetics

We describe a rare case of intradural-extramedullary primary spinal cysticercosis. A 42-year-old man visited our institute for lower back pain. He denied having consumed raw meet. Magnetic resonance (MR) images revealed an intradural pure cystic mass at the L3-L4 level. A radiologic diagnosis of spinal arachnoid cyst was established. Three years later, he complained of aggravated back pain, and follow-up MR examination showed a markedly expanded cyst, occupying the subarachnoid space from the T11 to the S1 level. L2 hemilaminectomy was performed, and a yellowish infected cyst bulged out through the dural opening. The cyst was removed en bloc. The histopathological findings of the cyst were consistent with parasitic infection. Serum enzyme-linked immunosorbent assay (ELISA) confirmed the presence of spinal cysticercosis. As there was no intracranial lesion, the final diagnosis was primary spinal cysticercosis, which is very rare. MR imaging is a sensitive diagnostic tool for detecting cystic lesions in the spine; however, it is difficult to distinguish cysticercosis from non-infectious cysts such as an arachnoid cyst without using gadolinium enhancement. Clinicians treating spinal cysts with an unusual clinical course should include cysticercosis as a differential diagnosis. We recommend contrast-enhanced MR imaging and serum ELISA in the diagnostic work-up of such cases.
Adult ; Arachnoid* ; Back Pain ; Cysticercosis* ; Diagnosis ; Diagnosis, Differential ; Enzyme-Linked Immunosorbent Assay ; Follow-Up Studies ; Gadolinium ; Humans ; Low Back Pain ; Magnetic Resonance Imaging ; Neurocysticercosis ; Spine ; Subarachnoid Space ; Taenia solium