Elevated levels of cyclophilin A secreted in milk during bovine mastitis.

Bovine mastitis is an inflammatory disease that primarily occurs when bacteria invade and proliferate in the mammary gland or such as physical trauma. Mastitis results in a decrease in milk yield and quality, causing huge economic losses. Cyclophilin A (CyPA) is a cytosolic protein known as cyclosporine binding protein. Recent studies have shown that CyPA is secreted from cells and has chemotactic activity, recruiting inflammatory cells and inducing multiple cytokines. In this study, we found that CyPA is detected in milk and is abundantly secreted at the onset of mastitis. A significant correlation was found between somatic cell counts (SCC) and the concentrations of CyPA in milk. To elucidate the relationship between mastitis and CyPA, we gave an intramammary infusion of S. aureus to cattle and investigated the attendant CyPA secretion. In S. aureus infused quarters, we observed an increased expression of CyPA on mammary epithelia and secretion into milk. The temporal profiles of CyPA in milk were synchronous with SCC, and there was a significant correlation between the concentration of CyPA in milk and SCC. These results suggest that CyPA is involved in the migration of immune cells during the onset of mastitis and may be used as a marker for the onset of mastitis.

Visualization of Recrystallization Induced by Ultraviolet Degradation of a Polypropylene Film Using Raman Imaging.

Raman images were constructed for polypropylene (PP) films before and after ultraviolet (UV) irradiation (100 mW, 248-436 nm) for 10 h using several intensity ratios of Raman bands that are sensitive to crystallization of PP. In the images of PP films before the irradiation the intensity ratios are nearly uniform for the films but for those of the PP films after the irradiation, the ratios become large with a mottled pattern, indicating that recrystallization occurs in the PP films upon the irradiation of the UV light. The UV-irradiated PP films show worm-like shaped structures in few micrometer order representing the recrystallization of PP. The temperature gradient of PP is low (273 K), and thus, it is very likely that due to UV energy and polymer fragmentation, PP molecules become more mobile and some parts of molecular chains in amorphous parts of PP molecules lead to their rearrangement and recrystallization. In this study, we demonstrate that Raman imaging clearly detects subtle changes in the crystallinity with a micrometer order structure which morphological images cannot observe.

Adjuvant multimodal treatment for spinal intradural extramedullary capillary hemangioma with subpial growth: illustrative case.

BACKGROUND: Spinal intradural extramedullary capillary hemangiomas are rare vascular lesions. Although total resection is the first treatment, the efficacy of adjuvant therapies, such as steroid or radiation therapy, has not been investigated. OBSERVATIONS: A 74-year-old man presented with progressive back pain, gait disturbance, and left chest pain. Spinal magnetic resonance imaging (MRI) revealed an intradural extramedullary lesion at the middle thoracic level. Preoperatively, systemic steroid administration improved his chest pain and reduced the tumor size. The authors performed tumor extirpation. Because the tumor strongly adhered to the pia matter, in short, the tumor accompanied with partial subpial growth, subtotal resection was performed, leaving a thin layer of the tumor. A pathological examination revealed capillary hemangioma. His gait disturbance showed rapid improvement. Stereotactic cyber knife treatment was performed for the residual tumor at 1 month after the operation. Serial MRI showed a gradual decrease in the tumor size and no recurrence at 2 years after the operation. LESSONS: When a tumor adheres to the spinal cord or nerve root, immoderate total resection should not be performed to avoid exacerbating the clinical symptoms. Adjuvant therapies, such as a systemic steroid administration or radiation therapy, may provide satisfactory control of spinal capillary hemangiomas.

Development of in-situ Raman diagnosis technique of eosinophil esophagitis.

The spectra of the live tissue with blood flow measured with 785 nm-excitation light showed a very weak signal due to hemoglobin (Hb). It suggested the possibility to detect eosinophil accumulation in the tissue with the 785 nm-excitation light. The excitation wavelength of 633 nm induced strong fluorescence of sapphire glass that is a material of the ball lens of BHRP (Ball lens top hollow optical fiber Raman probe). On the other hand, the previous study suggested that eosinophil including eosinophil peroxidase (EPO) that showed a strong resonance Raman effect with 633 nm-excitation light. The purpose of the present study is to collect basic information and to evaluate the viability of Raman spectroscopic analysis for the detection of eosinophil accumulation in the live esophagus. BHRP with a sapphire ball lens with 500 µm diameter was applied for measurement of live esophagus tissue of a mouse. In this study, Raman spectra of eosinophil were measured with 633 and 785 nm-excitation. The Raman spectra of eosinophil showed a strong contribution of EPO, suggested that a heme chromophore in EPO had pre-resonance enhancement via Q band with the 785 nm-excitation light. Principal component analysis (PCA) is applied for the analysis of Raman spectra of eosinophil, erythrocyte and other granulocytes. Eosinophil was successfully discriminated from other blood cells in the PCA score plots built for the datasets of the spectra measured with 633 and 785 nm-excitation wavelengths. Consequently, our study demonstrates that Raman spectroscopy with 785 nm-excitation had high viability for in situ analysis of eosinophilic esophagitis (EoE).

Prevention of mastitis in multiparous dairy cows with a previous history of mastitis by oral feeding with probiotic Bacillus subtilis.

Mastitis is a very common inflammatory disease of the mammary gland of dairy cows, resulting in a reduction of milk production and quality. Probiotics may serve as an alternative to antibiotics to prevent mastitis, and the use of probiotics in this way may lessen the risk of antibiotic resistant bacteria developing. We investigated the effect of oral feeding of probiotic Bacillus subtilis (BS) C-3102 strain on the onset of mastitis in dairy cows with a previous history of mastitis. BS feeding significantly decreased the incidence of mastitis, the average number of medication days and the average number of days when milk was discarded, and maintained the mean SCC in milk at a level substantially lower than the control group. BS feeding was associated with lower levels of cortisol and TBARS and increased the proportion of CD4+ T cells and CD11c+ CD172ahigh dendritic cells in the blood by flow cytometry analysis. Parturition increased the migrating frequency of granulocytes toward a milk chemoattractant cyclophilin A in the control cows, however, this was reduced by BS feeding, possibly indicating a decreased sensitivity of peripheral granulocytes to cyclophilin A. These results reveal that B. subtilis C-3102 has potential as a probiotic and has preventative capacity against mastitis in dairy cows.

Infantile-Onset Isolated Neurohypophyseal Langerhans Cell Histiocytosis with Central Diabetes Insipidus: A Case Report.

Central diabetes insipidus (CDI) is a rare disease in children and has a variety of etiologies. The major causes of CDI with pituitary stalk thickening (PST) are germinoma, Langerhans cell histiocytosis (LCH), and Lymphocytic infundibulo-neurohypophysitis, which are difficult to differentiate by imaging and require pathological diagnosis. We report a case of infantile-onset isolated neurohypophyseal LCH diagnosed by pathological findings. A 2-year-old girl presented with polydipsia and polyuria. CDI was diagnosed and treatment with oral desmopressin was initiated. Magnetic resonance imaging (MRI) of the head showed PST and absence of high-signal intensity of posterior pituitary on T1-weighted images. Follow-up MRI scans showed that the tumor mass was gradually increasing and extending posteriorly toward the area near the mamillary body. Simultaneously, anterior pituitary dysfunction was observed. She underwent a biopsy of the PST and LCH was diagnosed by immunohistochemical analysis. DNA analysis showed no BRAF V600E mutation. Monotherapy with 2-Chlorodeoxyadenosine reduced the tumor size but did not improve pituitary function. Isolated neurohypophyseal LCH should be considered in infantile-onset cases of CDI with PST. 2-CdA treatment resulted in rapid PST shrinkage. Further cases are needed to determine whether early diagnosis and treatment can prevent anterior pituitary dysfunction.

Attenuated Total Reflection Far-Ultraviolet (ATR-FUV) Spectroscopy is a Sensitive Tool for Investigation of Protein Adsorption.

Attenuated total reflection far-ultraviolet (ATR-FUV) spectra in the 145-250 nm region were studied for four kinds of proteins (two α-helix-rich proteins: bovine serum albumin (BSA) and lysozyme and two ß-sheet rich proteins: concanavalin A and γ-globulin) in different solutions (pure water and phosphate buffered saline, or PBS) with different concentrations. All the spectra show a band at 191 nm due to the π-π* transition of amide bonds of the proteins. The wavelength of the band does not change with their second structures, suggesting that the corresponding electronic transition mode is localized and polarized in the direction that is not affected by the difference in the peptide folding. The intensity of the 191 nm band differs with the concentration of salt in the solution, suggesting that the band intensity reflects the adsorption density of a protein on the internal reflection element (IRE) made of a sapphire glass prism. According to the intensity changes of the band at 191 nm, it is revealed that the properties in adsorption are different from one protein to another. It is assumed that there are two types of forces on the protein adsorption: one is that among the molecules and the other is that between a molecule and a substrate. The origin of force includes localized electrostatic polarity and affinity to water. The ions in the solvent give a marked effect on these forces, resulting in the difference in the response to adsorption density against the salt concentration in the solvent.

Simple Technique to Place the Lateral Mass Screws in the Revision Surgery after Cervical Laminoplasty for Ossification of the Posterior Longitudinal Ligament.

A lateral mass screw (LMS) is one of the standard anchor screws in posterior cervical fixation. Although the advantage of cervical LMS is that it is easier and safer to place than pedicle screw, it is sometimes difficult for surgeons to confirm the exact point for screw entry and accurate angle in cases of revision surgery. When LMS fixation is performed as revision surgery after cervical laminoplasty or laminectomy, it might be complicated to secure safe placement of the LMSs. We present a simple but practical technique involving a caliper and angle device for revision surgery after cervical laminoplasty for ossification of the posterior longitudinal ligament. In this technique, the distance between the bilateral entry points is ascertained using preoperative CT. Insertion of the screw is guided using the angle device set to 25 degrees. The technique presented here is easy and allows accurate placement of the LMSs in the posterior cervical spine, and is practical even for revision surgery.

Raman Study on Lipid Droplets in Hepatic Cells Co-Cultured with Fatty Acids.

The purpose of the present study was to investigate molecular compositions of lipid droplets changing in live hepatic cells stimulated with major fatty acids in the human body, i.e., palmitic, stearic, oleic, and linoleic acids. HepG2 cells were used as the model hepatic cells. Morphological changes of lipid droplets were observed by optical microscopy and transmission electron microscopy (TEM) during co-cultivation with fatty acids up to 5 days. The compositional changes in the fatty chains included in the lipid droplets were analyzed via Raman spectroscopy and chemometrics. The growth curves of the cells indicated that palmitic, stearic, and linoleic acids induced cell death in HepG2 cells, but oleic acid did not. Microscopic observations suggested that the rates of fat accumulation were high for oleic and linoleic acids, but low for palmitic and stearic acids. Raman analysis indicated that linoleic fatty chains taken into the cells are modified into oleic fatty chains. These results suggest that the signaling pathway of cell death is independent of fat stimulations. Moreover, these results suggest that hepatic cells have a high affinity for linoleic acid, but linoleic acid induces cell death in these cells. This may be one of the causes of inflammation in nonalcoholic fatty liver disease (NAFLD).

Re-Evaluation of the Prevalence of Permanent Congenital Hypothyroidism in Niigata, Japan: A Retrospective Study.

Although newborn screening (NBS) for congenital hypothyroidism (CH) in Japan started more than 40 years ago, the prevalence of CH remains unclear. Prevalence estimations among NBS-positive CH individuals include those with transient hypothyroidism and transient hyperthyrotropinemia, and re-evaluation with increasing age is necessary to clarify the actual incidence. Thus, we re-evaluated the incidence of permanent CH. Of the 106,114 patients who underwent NBS in the Niigata Prefecture, Japan, between April 2002 and March 2006, 116 were examined further due to high thyroid-stimulating hormone levels (>8 mIU/L) and were included in the study. We retrospectively evaluated their levothyroxine sodium (LT4) replacement therapy status from the first visit to 15 years of age. Of the 116 NBS-positive patients, 105 (91%) were initially examined in our department. Of these, 72 (69%) started LT4 replacement therapy on the first visit. Subsequently, 27 patients continued LT4 replacement until 15 years of age after multiple re-evaluations. The prevalence of permanent CH in the Niigata Prefecture during this period was 1 in 2500-3500 children. Ultimately, 62.5% of patients on LT4 replacement discontinued treatment by 15 years of age. This is the first study to clarify the true prevalence of permanent CH in Japan.

Coherent Anti-Stokes Raman Scattering Spectroscopy Using a Double-Wavelength-Emission Electronically Tuned Ti:Sapphire Laser.

Coherent anti-Stokes Raman scattering (CARS) spectroscopy is a powerful tool for Raman imaging technology. In contrast, conventional spontaneous Raman spectroscopy is often used for biological analysis with multivariate analysis. This study develops a new type of CARS instrument with a double-wavelength-emission, background-free, electronically tuned Ti:sapphire laser (DW-ETL). DW-ETL generates two laser pulses with different wavelengths simultaneously within its single resonator. The pulse wavelength and buildup time are regulated by acousto-optical tunable filter in the resonator. The present DW-ETL CARS system is free from any mechanical movement to measure a CARS spectrum by controlling each laser pulse of the emission throughout the fingerprint region. Consequently, it is theoretically able to provide stable CARS spectra to apply multivariate analysis in biological applications. The present study demonstrates that the DW-ETL CARS system provides spectra of biomedical samples in the full finger-print region, and the stability and controllability of the system are evaluated.

Intramedullary spinal cord metastasis of gastric cancer.

The incidence of intramedullary spinal cord metastasis (ISCM) has been increasing because the overall survival of patients with cancer has improved thanks to recent advanced therapies, such as molecular targeted drugs, anticancer agents, and various irradiation techniques. ISCM from lung and breast cancer is the most common form among cases of ISCM. We report an extremely rare form of ISCM from gastric cancer. This 83-year-old man who had a past medical history of gastric adenocarcinoma presented with acute onset of paraparesis. Spinal magnetic resonance imaging revealed an intramedullary lesion at the upper thoracic level. Due to rapid worsening of his paresis, we decided to perform tumor extirpation. Gross total resection of the tumor was successfully performed. Pathological examination revealed poorly differentiated adenocarcinoma, suggesting the diagnosis of ISCM from gastric cancer. He demonstrated gradual improvement of paraparesis soon after surgery, although his overall survival was limited to about 6 months after surgery. When examining the etiology of acute paraparesis in elderly patients with a past medical history of cancer, ISCM should be considered in the differential diagnosis. The prognosis of ISCM from gastric cancer is still extremely limited. Unfortunately, there is currently no treatment with proven efficacy. Surgery for ISCM from gastric cancer, although a challenging procedure for spine surgeons, should be considered as a therapeutic option in these patients.

Study of Cold-Pressed Tobacco Seed Oil Properties by Gas Chromatography Method.

A special place among vegetable oils was occupied by natural tobacco oil. Natural tobacco oil in folk medicine is used as an antiseptic and antibacterial agent. To determine the possible alternative use of tobacco, the seeds representing Kazakhstan tobacco cultivars, extracted from ground Nicotiana tabacum seeds, were investigated by using the cold press. The quality of the oil was assessed in terms of free fatty acid content. The percentage oil yield was 36.75 ± 0.50%. Tobacco seed oil is highly unsaturated, nonacidic, and would require little purification. The oil can be used for the production of soap, antiwrinkle cream for the face, hair shampoo, shoe cream, and as a high-resolution base oil for perfumes and light industry and can be commercialized due to its high oil yield, for use as anti-inflammatory drugs.

Complex Revision Surgery for Cervical Deformity or Implant Failure.

Postoperative cervical deformity sometimes occurs in the short or long term after primary surgery for cervical disorders related to the degenerative aging spine, neoplastic etiologies, hemodialysis, infection, inflammation, trauma, etc. Cervical kyphosis after posterior decompression surgery, such as laminectomy or laminoplasty, is a common problem for spine surgeons. However, revision surgery for cervical deformity is definitely one of the most challenging areas for spine surgeons. There is no doubt that surgery for cervical deformity carries a high risk of surgery-related complications that might result in aggravation of healthrelated quality of life. Revision surgery is even more challenging. Hence, spine surgeons need to assess carefully the overall severity of the underlying condition before revision surgery, and try to refine the surgical strategy to secure safe surgery. Needless to say, spine surgeons are now facing great challenges in making spine surgery a much more reliable and convincing entity.

Timing of hyponatremia development in patients with salt-wasting-type 21-hydroxylase deficiency.

Newborn screening (NBS) can detect 21-hydroxylase deficiency (21-OHD), allowing for early treatment initiation. However, many patients present with adrenal crises or hyponatremia at their first visit. Age (in days) of hyponatremia development in infants with salt-wasting (SW)-type 21-OHD remains unclear. Therefore, we determined the earliest age of hyponatremia diagnosis in this retrospective observational study using medical records of 40 patients with classic 21-OHD in Niigata Prefecture, Japan, from April 1989 to March 2019. We determined the earliest diagnosis of hyponatremia (serum sodium levels < 130 mEq/L) and created a sodium decrease rate model to estimate hyponatremia development age. Of 23 patients with SW-type 21-OHD, 10 (43.5%) were identified during NBS; the earliest case to present with hyponatremia was at day 7. Serum sodium levels were significantly and negatively correlated with age in days, and hyponatremia was estimated to develop at 6.6 d after birth. Genotype or serum 17-hydroxyprogesterone levels were not associated with sodium decrease rate. Thus, hyponatremia development age is earlier (within 7 d) than the previously described time-point (10-14 d) in infants with SW-type 21-OHD. Efforts to reduce the time lag from obtaining results to consultation may be required in patients with high 17-hydroxyprogesterone levels on NBS.

Review of Radiological Parameters, Imaging Characteristics, and Their Effect on Optimal Treatment Approaches and Surgical Outcomes for Cervical Ossification of the Posterior Longitudinal Ligament.

Determining the optimal surgical method for cervical ossification of the posterior longitudinal ligament (OPLL) is challenging. The surgical indication should be made based on not only radiological findings, but also the patient's age, preoperative neurological findings, social background, activities of daily life, and the presence or absence of comorbid diseases. Anterior resection for OPLL with or without wide corpectomy and fusion, posterior decompression with or without relatively long fusion, or anterior and posterior combined surgery may be considered. When evaluating the clinical condition of patients with cervical OPLL before surgery, various radiological parameters should be carefully considered, including the number of spinal segments involved, the cervical alignment or tilt angle, the relationship between OPLL and the C2-7 line (termed the "K-line"), the occupying ratio of OPLL, and the involvement of dural ossification. The objective of this article is to review the radiological parameters in current use for deciding upon the optimal surgical strategy and for predicting surgical outcomes, focusing on cervical OPLL.

Familial Os Odontoideum: Proatlas Segmentation Abnormality.

BACKGROUND: Os odontoideum is a smooth, independent ossicle separated from a hypoplastic odontoid process located cranially in the expected position of the odontoid tip. The pathogenesis of os odontoideum remains controversial, and several etiological theories have been suggested for congenital, developmental, and traumatic origins. CASE DESCRIPTION: We have reported symptomatic familial cases of dystopic os odontoideum in 2 elderly sisters who denied any traumatic history of the head and neck. Both patients were treated surgically, with successful outcomes achieved. CONCLUSIONS: When examining the etiology of os odontoideum, checking for the existence of a traumatic history is important. However, a more important matter is to consider morphological segmentation abnormalities of the proatlas. Segmentation abnormalities of the proatlas could be closely connected to the development of familial os odontoideum.

Early detection of virus infection in live human cells using Raman spectroscopy.

Virus infection of a human cell was determined only 3 h after invagination. We used viral vector Ad-CMV-control (AdC), which lacks the E1 gene coding for early polypeptide 1 (E1). AdC can replicate in human embryonic kidney 293 (HEK293) cells into which the E1 gene has been transfected. According to partial least-square regression discriminant analysis, it was assumed that two kinds of reaction take place in the cell during viral invasion. The first response of the cell was determined 3 h after the virus invasion, and the second one was determined ∼9 h later. The first one seems to be due to compositional changes in DNA. Analysis of large-scale datasets strongly indicated that the second reaction can be attributed to a reduction in protein concentration or uptake of phenylalanine into the nucleus.

Phenotypic and functional analysis of bovine peripheral blood dendritic cells before parturition by a novel purification method.

Dendritic cells (DCs) are specialized antigen presenting cells specializing in antigen uptake and processing, and play an important role in the innate and adaptive immune response. A subset of bovine peripheral blood DCs was identified as CD172a+ /CD11c+ /MHC (major histocompatibility complex) class II+ cells. Although DCs are identified at 0.1%-0.7% of peripheral blood mononuclear cells (PBMC), the phenotype and function of DCs remain poorly understood with regard to maintaining tolerance during the pregnancy. All cattle used in this study were 1 month before parturition. We have established a novel method for the purification of DCs from PBMC using magnetic-activated cell sorting, and purified the CD172a+ /CD11c+ DCs, with high expression of MHC class II and CD40, at 84.8% purity. There were individual differences in the expressions of CD205 and co-stimulatory molecules CD80 and CD86 on DCs. There were positive correlations between expression of cytokine and co-stimulatory molecules in DCs, and the DCs maintained their immune tolerance, evidenced by their low expressions of the co-stimulatory molecules and cytokine production. These results suggest that before parturition a half of DCs may be immature and tend to maintain tolerance based on the low cytokine production, and the other DCs with high co-stimulatory molecules may already have the ability of modulating the T-cell linage.

Discrimination analysis of excitatory and inhibitory neurons using Raman spectroscopy.

We have succeeded in discriminating between intact excitatory and inhibitory neuronal cells with Raman analysis. Excitatory and inhibitory neurons have several differences in their electric activities, but it can be difficult to determine their types based only on visual appearances. As Raman spectroscopy does not require any staining or labeling, its use in live neuronal cells is possible. In the present study, we used primary neurons obtained from rat cerebral cortexes, which we cultured on a glial feeder layered culturing dish for 15 days. The Raman spectra of the intact neurons on the dish were obtained; the neurons were then immunostained and their types determined. Partial least squares regression-discriminant analysis (PLSR-DA) was employed for classification of the excitatory and inhibitory neurons. The results demonstrated a high feasibility for use of Raman spectroscopy for discrimination analysis of inhibitory and excitatory neurons in a nondestructive manner.