著者

Mami ADACHI Yuki HOSHINO Yusuke IZUMI Satoshi TAKAGI

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0625, (Released:2015-12-21)

被引用文献数

1 8

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Canine hemangiosarcoma (HSA) is a progressive malignant neoplasm of dogs for which there is currently no effective treatment. A recent study suggested that receptor tyrosine kinases (RTKs), the PI3K/Akt/m-TOR and MAPK pathways are all activated in canine and human HSA. The aim of the present study was to investigate the overexpression of these proteins by immunohistochemistry in canine splenic HSA to identify potential molecular therapeutic targets. A total of 10 splenic HSAs and two normal splenic samples surgically resected from dogs were sectioned and stained with hematoxylin and eosin for histological diagnosis or analyzed using immunohistochemistry. The expression of RTKs, c-kit, VEGFR-2 and PDGFR-2, as well as PI3K/Akt/m-TOR and MEK was higher in canine splenic HSAs compared to normal spleens. These proteins may therefore be potential therapeutic targets in canine splenic HSA.

著者

大沼 学 鈴木 正嗣 内田 英二 新山 雅美 大泰司 紀之

出版者

社団法人日本獣医学会

雑誌

The journal of veterinary medical science (ISSN:09167250)

巻号頁・発行日

vol.64, no.4, pp.s・vii, 309-313, 2002-04-25

被引用文献数

1 12

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1998年8月から1999年7月にかけて,3頭の非妊娠,雌マレーグマ(Helarctos malayanus)を対象に糞中から検出されるエストラジオールの定量を,マレイシア国サラワク州において実施した.また,それに加えて膣粘液中に見出される細胞の観察を1998年8月と1999年3月にマレーグマ1頭を対象に行った.3頭の糞中エストラジオール濃度は,1998年8月または9月にピークが観察された.また,膣粘液中に観察される角化上皮細胞の出現割合は,1998年8月の数値が1999年3月のものより高かった.これらのことからマレイシア国サラワク州において,マレーグマはエストラジオールの濃度がピークをむかえる8月から9月ごろに発情している可能性が示唆された.

著者

Maho URATA Rie WATANABE Hiroyuki IWATA

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

vol.77, no.12, pp.1611-1616, 2015 (Released:2016-01-01)

参考文献数

28

被引用文献数

2

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The cytotoxicity of Ibaraki virus nonstructural protein NS3 was confirmed, and the contribution of glycosylation to this activity was examined by using glycosylation mutants of NS3 generated by site-directed mutagenesis. The expression of NS3 resulted in leakage of lactate dehydrogenase to the culture supernatant, suggesting the cytotoxicity of this protein. The lack of glycosylation impaired the transport of NS3 to the plasma membrane and resulted in reduced cytotoxicity. Combined with the previous observation that NS3 glycosylation was specifically observed in mammalian cells (Urata et al., Virus Research 2014), it was suggested that the alteration of NS3 cytotoxicity through modulating glycosylation is one of the strategies to achieve host specific pathogenisity of Ibaraki virus between mammals and vector arthropods.

著者

Mijung KIM Yongsun KIM Seunghoon LEE Minyoung KUK Ah Young KIM Wanhee KIM Oh-Kyeong KWEON

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0361, (Released:2015-12-27)

被引用文献数

6

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Allogenic adipose-derived mesenchymal stem cells (Ad-MSCs) are an alternative source for cytotherapy owing to their antioxidant and anti-inflammatory effects. Frozen-thawed allogenic Ad-MSCs can be used instantly for this purpose. However, the viability and function of frozen-thawed Ad-MSCs have not been clearly evaluated. The purpose of this study was to compare the viability and function of Ad-MSCs and heme oxygenase-1 (HO-1)-overexpressed Ad-MSCs in vitro after freeze-thawing. The viability, proliferation, antioxidant capacity and mRNA gene expression of growth factors were evaluated. Frozen-thawed cells showed significantly lower viability than fresh cells (77% for Ad-MSCs and 71% for HO-1 Ad-MSCs, P<0.01). However, the proliferation rate of frozen-thawed Ad-MSCs increased and did not differ from that of fresh Ad-MSCs after 3 days of culture. In contrast, the proliferation rate of HO-1-overexpressed Ad-MSCs was lower than that of Ad-MSCs. The mRNA expression levels of TGF-β, HGF and VEGF did not differ between fresh and frozen-thawed Ad-MSCs, but COX-2 and IL-6 had significantly higher mRNA expression in frozen cells than fresh cells (P<0.05). Fresh Ad-MSCs exhibited higher HO-1 mRNA expression than frozen-thawed Ad-MSCs, and fresh HO-1 overexpressed Ad-MSCs exhibited higher than fresh Ad-MSCs (P<0.05). However, there was no significant difference between fresh and frozen HO-1 overexpressed Ad-MSCs. The antioxidant capacity of HO-1-overexpressed Ad-MSCs was significantly higher than that of Ad-MSCs. Cryopreservation of Ad-MSCs negatively affects viability and antioxidant capacity, and HO-1-overexpressed Ad-MSCs might be useful to maximize the effect of Ad-MSCs for cytotherapy.

著者

Jongsu LIM Youngmin YOON Dongin JUNG Seongchan YEON Heechun LEE

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0181, (Released:2015-12-12)

被引用文献数

2

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A 16-month-old intact female Maltese dog was referred for examination of depression and vomiting. Ultrasonography revealed dilated right renal pelvis containing echogenic fluid with free gas. A hyperechoic material suspected of urolith was identified in the right ureter. Computed tomography revealed emphysematous change of the right kidney associated with ureteral obstruction and extrahepatic portosystemic shunt (EHPSS). Ureteronephrectomy and surgical correction were performed for the EHPSS. Escherichia coli was isolated from pus from the right kidney. Quantitative analysis revealed that the urolith was an ammonium urate stone. After 5 months follow-up, no complication was observed. This is the first report of emphysematous pyonephrosis associated with EHPSS in a dog.

著者

Atigan THONGTHARB Kazuyuki UCHIDA James K. CHAMBERS Yumiko KAGAWA Hiroyuki NAKAYAMA

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0627, (Released:2015-12-12)

被引用文献数

26

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Histiocytic sarcoma is a progressive and fatal malignant neoplasm that mainly occurs in middle- to old-aged dogs. This study describes clinicopathological, histological and immunohistochemical characteristics of intracranial histiocytic sarcomas in 23 dogs. Magnetic resonance imaging and/or computed tomography of the brains revealed that the tumors mainly located in the cerebrum, particularly the frontal lobe. Seizure was a predominant clinical sign in most of the cases. Histologically, the tumor cells were morphologically classified into round/polygonal- and spindle-shaped cell types. There was a significant association between tumor cell types and hemophagocytic activity (P<0.05). However, there was no significant difference in other clinicopathological parameters and mitotic index between the 2 types. Immunohistochemically, tumor cells were strongly positive for HLA-DR, Iba-1 and CD204 in all the 23 cases, for iNOS in 20, for CD163 in 17, for lysozyme and CD208 (DC-LAMP) in 9, and for S100 in 5 cases. In addition, the Ki67-proliferative index showed range of 0.50–64.33% (Average 26.60 ± 3.81%). These observations suggest that canine primary intracranial histiocytic sarcomas tend to exhibit both dendritic cell and macrophage phenotypes of histiocytic differentiation.

著者

Yosuke ITO Seiya MAEHARA Yoshiki ITOH Ai MATSUI Miri HAYASHI Akira KUBO Tsuyoshi UCHIDE

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0227, (Released:2015-12-08)

被引用文献数

6

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The purpose of this study was to investigate the effects of refractive error on canine visual evoked potentials with pattern stimulation (P-VEP). Six normal beagle dogs were used. The refractive power of the recorded eyes was measured by skiascopy. The refractive power was corrected to −4 diopters (D) to +2 D using contact lens. P-VEP was recorded at each refractive power. The stimulus pattern size and distance were 50.3 arc-min and 50 cm. The P100 appeared at almost 100 msec at −2 D (at which the stimulus monitor was in focus). There was significant prolongation of the P100 implicit time at −4, −3, 0 and +1 D compared with −2 D, respectively. We concluded that the refractive power of the eye affected the P100 implicit time in canine P-VEP recording.

著者

Yasuyuki KANEKO Shidow TORISU Mitsuyoshi HAGIO Ryoji YAMAGUCHI Shinya MIZUTANI Kiyokazu NAGANOBU

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0425, (Released:2015-11-30)

被引用文献数

2

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Hepatopulmonary syndrome (HPS) is a respiratory complication of hepatic disease, that is well recognized in humans and defined by the presence of 1) liver disease, 2) hypoxemia and/or high alveolar-arterial oxygen gradient (AaDO2) and 3) intrapulmonary vasodilatation. The present report describes a similar case of HPS in a dog. A six-month-old Papillon was diagnosed with ductal plate malformation with chronic active hepatitis and showed progressive increases in AaDO2 over the course of the following six months. The presence of intrapulmonary vasodilatation was confirmed by agitated saline contrast transthoracic echocardiography. Also, the absence of congenital cardiac defect was confirmed by transthoracic echocardiography. From these results, we suspected that this dog had HPS. This is the first description of suspected canine HPS.

著者

Masaki MICHISHITA Aya OHTSUKA Rei NAKAHIRA Tsuyoshi TAJIMA Takayuki NAKAGAWA Nobuo SASAKI Toshiro ARAI Kimimasa TAKAHASHI

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0550, (Released:2015-11-28)

被引用文献数

1 22

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Feline mammary carcinomas are characterized by rapid progression and metastases. Vascular endothelial growth factor (VEGF) is a key regulator of tumor angiogenesis, proliferation and metastasis. The present study aimed to investigate the effects of a single drug therapy of bevacizumab on a xenograft model of feline mammary carcinoma expressing VEGF protein. Bevacizumab treatment suppressed tumor growth by inhibiting angiogenesis and enhancing apoptosis; however, it did not affect the tumor proliferation index. Thus, bevacizumab had anti-tumor effects on a xenograft model, and this may be useful for the treatment of feline mammary carcinoma.

著者

Soohyun KIM Young Woo PARK Euiri LEE Sang Wan PARK Sungwon PARK Hyunwoo NOH Jong Whi KIM Je Kyung SEONG Kangmoon SEO

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0455, (Released:2015-11-26)

被引用文献数

5

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This study evaluated the effect of onion extract on corneal haze suppression after applying the air assisted lamellar keratectomy. The air assisted lamellar keratectomy was performed on 24 canine eyes. They were treated with an artificial tear (group C), prednisolone acetate (group P), onion extract (group O) and TGF-β1 (group T) three times per day from 7 to 28 days after the surgery. Corneal haze occurred on the all eyes and was observed beginning 7 days after the surgery. The haze was significantly decreased in groups P and O from day 14 compared with the group C using the clinical (group P; P=0.021, group O; P=0.037) and objective evaluation method (group P; P=0.021, group O; P=0.039). In contrast, it was significantly increased in group T from day 14 compared with group C based on the clinical (P=0.002) and objective evaluation method (P<0.001). Subsequently, these eyes were enucleated after euthanasia, and immunohistochemistry with α-SMA antibodies was done. The total green intensity for α-SMA was significantly more expressed in group T and significantly less expressed in groups P and O than in group C. Onion extract could have potential as a therapeutic in preventing corneal haze development by suppressing the differentiation of fibroblasts into myofibroblasts.

著者

Michihito TAGAWA Eiichi KANAI Genya SHINBOU Mikiya KANO Hideki KAYANUMA

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0456, (Released:2015-11-21)

被引用文献数

9

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Depth to width ratio (D/W) is the only quantitative item in the criteria recommended by the Japanese Ultrasound Society for the evaluation of breast tumors in humans. However, the usefulness of the D/W has not been evaluated in dogs. Eighty-six mammary masses in 34 female dogs underwent ultrasonographic examination to determine the D/W and other characteristics. Results of ultrasonographic and histopathologic examinations were compared. The D/W of malignant tumors was significantly greater than that of benign tumors, and it had a sensitivity of 56.3% and a specificity of 92.9% for the diagnosis of malignancy when the threshold of D/W was 0.7. In addition, irregular margin, polymorphous shape and heterogeneous internal echographic characteristics were correlated with malignancy.

著者

Sayaka ARATA Ai ASAHI Yukari TAKEUCHI Yuji MORI

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0579, (Released:2015-11-03)

被引用文献数

3

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Eighteen autosomal microsatellite loci were examined using 275 Shiba Inus in Japan. Eighteen dogs representing eight trios were obtained from four breeders to calculate mutation rates, and 257 dogs kept by owners were collected through veterinary clinics throughout Japan to calculate population genetic parameters and estimate discrimination power. After two loci (INU005 and AHTk253) were excluded, average expected heterozygosity (He), polymorphic information content (PIC), and fixation index (F) were 0.665, 0.623, and 0.046, respectively. The combined power of discrimination over the 16 microsatellite markers was more than 0.9999. Therefore, it is suggested that these 16 microsatellite loci recommended by the International Society for Animal Genetics (ISAG) are applicable for individual identification and parentage testing of domestic Shiba Inu in Japan.

著者

Moeko KOHYAMA Naomi TADA Hiroko MITSUI Hitomi TOMIOKA Toshihiko TSUTSUI Akira YABUKI Mohammad Mahbubur RAHMAN Kazuya KUSHIDA Keijiro MIZUKAMI Osamu YAMATO

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0279, (Released:2015-11-06)

被引用文献数

1 11

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Canine progressive rod-cone degeneration (PRCD) is a middle- to late-onset, autosomal recessive, inherited retinal disorder caused by a substitution (c.5G>A) in the canine PRCD gene that has been identified in 29 or more purebred dogs. In the present study, a TaqMan probe-based real-time PCR assay was developed and evaluated for rapid genotyping and large-scale screening of the mutation. Furthermore, a genotyping survey was carried out in a population of the three most popular breeds in Japan (Toy Poodles, Chihuahuas and Miniature Dachshunds) to determine the current mutant allele frequency. The assay separated all the genotypes of canine PRCD rapidly, indicating its suitability for large-scale surveys. The results of the survey showed that the mutant allele frequency in Toy Poodles was high enough (approximately 0.09) to allow the establishment of measures for the prevention and control of this disorder in breeding kennels. The mutant allele was detected in Chihuahuas for the first time, but the frequency was lower (approximately 0.02) than that in Toy Poodles. The mutant allele was not detected in Miniature Dachshunds. This assay will allow the selective breeding of dogs from the two most popular breeds (Toy Poodle and Chihuahua) in Japan and effective prevention or control of the disorder.

著者

Karin UESAKA Masaki MAEZAWA Hisashi INOKUMA

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0392, (Released:2015-10-31)

被引用文献数

4

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A serological survey of Borrelia infection of dogs was performed in Sapporo, Japan, where Borrelia garinii infection in dogs was detected in 2011. A total of 314 serum samples were collected from dogs that visited three animal hospitals in Sapporo from 2012 to 2014. The two-step evaluation method, involving screening ELISA followed by Western blot analysis, was used to detect antibodies against Borrelia species. A total of 34 samples were positive by ELISA. Among those 34 samples, 32 were positive for Borrelia spp. by Western blot. These findings suggest that the 32 dogs (10.2%) generated antibodies against Borrelia burgdorferi sensu lato, such as B. garinii or B. afzelii. Antibody positivity was 7.6% and 13.3% for dogs living in urban and rural areas, respectively. Dogs with a history of tick infestation showed a positive rate of 16.7%, which was higher, although not significantly, than the 6.7% among dogs without a history.

著者

Melanie HAAS Franz-Josef KAUP Stephan NEUMANN

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0415, (Released:2015-10-31)

被引用文献数

1 19

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Interleukin-8 (IL-8 or CXCL8) is a highly selective pro-inflammatory chemokine, that is elevated in sera of humans and animals with various inflammatory diseases. CXCL8 is possibly involved in uncontrolled inflammation and the development of a systemic inflammatory response syndrome (SIRS) and sepsis. Nevertheless, its behavior and precise properties in the course of inflammation are not fully understood. Thus, we used naturally occurring canine pyometra as a model of inflammation, in order to examine the behavior of serum CXCL8 in relation to the disease intensity and commonly analyzed inflammatory mediators. Using a commercially available canine ELISA kit, a significant increase of CXCL8 was determined in the serum of 23 dogs with pyometra compared with 35 healthy dogs. Interestingly, serum CXCL8 did not increase in severely diseased patients and behaved contrary to white blood cells (WBC), neutrophils and C-reactive protein (CRP). The measurement of serum CXCL8 may provide valuable information about the extent of ongoing lesions and could be a useful complement for existing laboratory tests.

著者

Nozomi SHIWA Hassadin BOONSRIROJ Kazunori KIMITSUKI Taichi SHIMATSU Chun-Ho PARK

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0467, (Released:2015-10-31)

被引用文献数

2

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A 15-year-old, female mixed-breed dog presented with abdominal distention. An exploratory laparotomy revealed a large left ovarian mass (20 × 15 × 12 cm). Histopathological examination of the mass revealed a mixed myxoid liposarcoma and a well-differentiated leiomyoma. Four months after surgical removal of the mass, the dog died due to multiorgan metastasis. The metastasis was composed solely of the liposarcoma component. The liposarcoma component was Alcian Blue- and Oil red O-positive, and demonstrated immunoreactivity with S-100, adipophilin and vimentin. Electron microscopy revealed that the tumor cell cytoplasms were packed with lipid vacuoles and dilated rough endoplasmic reticulum. To our knowledge, this is the first report of myxoid liposarcoma and leiomyoma co-existing in a canine ovary.

著者

Domenico CAIVANO Francesco BIRETTONI Antonello BUFALARI Valentina DE MONTE Giovanni ANGELI Maria Elena GIORGI Valentina PATATA Francesco PORCIELLO

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0417, (Released:2015-10-24)

被引用文献数

5

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Lung lobe torsion is rare but life-threatening condition in the dog. Thoracic radiographs and conventional ultrasonography cannot be conclusive for the diagnosis, and computed tomography is useful but is limited by cost and availability. This report describes the findings of contrast-enhanced ultrasonography in three dogs with lung lobe torsion. Contrast-enhanced ultrasonography showed the absence or reduction of pulmonary vascularization secondary to twisting of the lung lobe around its bronchovascular pedicle in all three dogs. Moreover, contrast-enhanced ultrasonography distinguished partial pulmonary atelectasis from a lung lobe torsion. These preliminary results suggest that contrast-enhanced ultrasonography can improve the accuracy of conventional ultrasonography for detection of pulmonary blood flow compromise in dogs with lung lobe torsion.

著者

Mai INOUE Atsuhiko HASEGAWA Yuta HOSOI Katsuaki SUGIURA

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0171, (Released:2015-10-12)

被引用文献数

4

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The association between breed, gender and age and cardiovascular disorders in the insured dog population in Japan was investigated, using multiple logistic regression analysis and data from 299,555 dogs insured between April 2010 and March 2011. The overall annual prevalence of cardiovascular disorder diagnosis was 2.1%. Using the Miniature Dachshund as the reference breed, Cavalier King Charles Spaniel had the highest odds of cardiovascular disorder with a ratio of 16.2 (95% confidence interval: 14.4–18.2), followed by Maltese, Pomeranian, Chihuahua and Shih Tzu. Male dogs had increased odds of 1.2 (1.1–1.3). The dogs had increased odds of having cardiovascular disorder by 1.5 times as their age increased by one year.

著者

Tomomi TAKANO Saya YAMASHITA Michiko MURATA-OHKUBO Kumi SATOH Tomoyoshi DOKI Tsutomu HOHDATSU

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0347, (Released:2015-10-11)

被引用文献数

7

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We collected rectal swabs from dogs in Japan during 2011 to 2014, and canine coronavirus (CCoV) nucleocapsid gene was detected by RT-PCR. The relationship between CCoV infection and the manifestation of diarrhea symptoms was investigated, and a correlation was noted (df=1, χ2=8.90, P<0.005). The types of CCoV detected in samples from CCoV-infected dogs were CCoV-I in 88.9% and CCoV-II in 7.4%, respectively. We retrospectively investigated the seroprevalence of CCoV-I in dogs in Japan during 1998 to 2006. The sera were tested with a neutralizing antibody test. In the absence of CCoV-I laboratory strain, we used feline coronavirus (FCoV)-I that shares high sequence homology in the S protein with CCoV-I. 77.7% of the sera were positive for neutralizing anti-FCoV-I antibodies.

著者

Kenjiro FUKUSHIMA Nozomi EGUCHI Koichi OHNO Hideyuki KANEMOTO Masashi TAKAHASHI Hirotaka IGARASHI Aki OHMI Ko NAKASHIMA Hajime TSUJIMOTO

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0129, (Released:2015-10-10)

被引用文献数

15

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Inflammatory colorectal polyp (ICRP), common in miniature dachshunds, presents with hematochezia, tenesmus and mucoid feces. Although an 80% response rate has been reported when treated with prednisolone and cyclosporine, effective treatment is needed for the remaining 20% of ICRP dogs. Leflunomide is an immunosuppressive drug reported as effective in several immune-mediated diseases. In the present study, we retrospectively evaluated the efficacy and adverse effects of leflunomide in 15 ICRP dogs that were refractory to treatment with prednisolone and cyclosporine. Treatment efficacy was assessed by endoscopy, clinical symptoms and rectal palpation. Adverse effects were determined by clinical symptoms and blood testing during follow-up. The leflunomide treatment response rate was 93.3%. The median dosage of leflunomide and the median response time were 3 mg/kg (range: 1.7–4.0 mg/kg) and 35 days (range: 20–119 days), respectively. Adverse effects observed included lethargy (3 dogs), anorexia (1 dog), respiratory symptoms (1 dog), leukocytopenia (2 dogs), thrombocytopenia (1 dog), anemia (1 dog), and liver enzyme elevation (8 dogs). Most of the adverse effects improved with symptomatic treatment and leflunomide discontinuation or dosage reduction. In conclusion, leflunomide treatment is effective in ICRP dogs refractory to treatment with prednisolone and cyclosporine. Because several adverse effects were observed, close monitoring is needed during leflunomide treatment follow-up.