CRISPR loss of function screening to identify genes involved in human primordial germ cell-like cell development.

Despite our increasing knowledge of molecular mechanisms guiding various aspects of human reproduction, those underlying human primordial germ cell (PGC) development remain largely unknown. Here, we conducted custom CRISPR screening in an in vitro system of human PGC-like cells (hPGCLCs) to identify genes required for acquisition and maintenance of PGC fate. Amongst our candidates, we identified TCL1A, an AKT coactivator. Functional assessment in our in vitro hPGCLCs system revealed that TCL1A played a critical role in later stages of hPGCLC development. Moreover, we found that TCL1A loss reduced AKT-mTOR signaling, downregulated expression of genes related to translational control, and subsequently led to a reduction in global protein synthesis and proliferation. Together, our study highlights the utility of CRISPR screening for human in vitro-derived germ cells and identifies novel translational regulators critical for hPGCLC development.

Structure of a HIV-1 IN-Allosteric inhibitor complex at 2.93 Å resolution: Routes to inhibitor optimization.

HIV integrase (IN) inserts viral DNA into the host genome and is the target of the strand transfer inhibitors (STIs), a class of small molecules currently in clinical use. Another potent class of antivirals is the allosteric inhibitors of integrase, or ALLINIs. ALLINIs promote IN aggregation by stabilizing an interaction between the catalytic core domain (CCD) and carboxy-terminal domain (CTD) that undermines viral particle formation in late replication. Ongoing challenges with inhibitor potency, toxicity, and viral resistance motivate research to understand their mechanism. Here, we report a 2.93 Å X-ray crystal structure of the minimal ternary complex between CCD, CTD, and the ALLINI BI-224436. This structure reveals an asymmetric ternary complex with a prominent network of π-mediated interactions that suggest specific avenues for future ALLINI development and optimization.

Effect of Sample Geometry on Graphitization of Polyacrylonitrile.

In this study, it is analyzed how sample geometry (spheres, nanofibers, or films) influences the graphitization behavior of polyacrylonitrile (PAN) molecules. The chemical bonding and changes in the composition of these three geometries are studied at the oxidation, carbonization, and graphitization stages via scanning electron microscopy (SEM), in situ thermogravimetric-infrared (TGA-IR) analysis, elemental analysis, Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS). The influence of molecular alignment on the graphitization of the three sample geometries is investigated using synchrotron wide-angle X-ray diffraction (WAXD) and transmission electron microscopy (TEM). The effects of molecular alignment at different draw rates during spinning are explored in detail.

Disruption of TET2 promotes the therapeutic efficacy of CD19-targeted T cells.

Cancer immunotherapy based on genetically redirecting T cells has been used successfully to treat B cell malignancies1-3. In this strategy, the T cell genome is modified by integration of viral vectors or transposons encoding chimaeric antigen receptors (CARs) that direct tumour cell killing. However, this approach is often limited by the extent of expansion and persistence of CAR T cells4,5. Here we report mechanistic insights from studies of a patient with chronic lymphocytic leukaemia treated with CAR T cells targeting the CD19 protein. Following infusion of CAR T cells, anti-tumour activity was evident in the peripheral blood, lymph nodes and bone marrow; this activity was accompanied by complete remission. Unexpectedly, at the peak of the response, 94% of CAR T cells originated from a single clone in which lentiviral vector-mediated insertion of the CAR transgene disrupted the methylcytosine dioxygenase TET2 gene. Further analysis revealed a hypomorphic mutation in this patient's second TET2 allele. TET2-disrupted CAR T cells exhibited an epigenetic profile consistent with altered T cell differentiation and, at the peak of expansion, displayed a central memory phenotype. Experimental knockdown of TET2 recapitulated the potency-enhancing effect of TET2 dysfunction in this patient's CAR T cells. These findings suggest that the progeny of a single CAR T cell induced leukaemia remission and that TET2 modification may be useful for improving immunotherapies.

Beauty Pageants in the Japanese Colonial Era of Korea (1931 and 1940).

The aim of this study was to analyze the beauty pageants that were held in Korea during the Japanese colonial era (1931, 1940). Through searches of www.google.com and www.naver.com, 3 pageants were found: (1) 1931, Miss Korea, organized by Samchunli (a magazine): Any Korean woman, with the exception of Korean geishas, could apply by submitting her photographs. The number of candidates was 326. Nine Korean judges evaluated the photos of the candidates. Among them, the victor was an 18-year-old Korean woman. (2) 1931 (Oct), Miss Joseon, organized by Osaka Mainichi Shimbun: The candidates were categorized into 2 groups: 10 "internals" (Japanese) and 10 Koreans. Their heights and weights, along with their photographs, were provided for the readers' reference. The winners were determined based on the readers' votes. The Japanese victor was a 21-year-old student with a height of 150 cm and a weight of 52 kg (body mass index [BMI] 23.11 kg/m2). The Korean winner was a 19-year-old student standing at 157 cm and weighing 53 kg (BMI 21.50 kg/m2). (3) 1940, Venus of the Peninsula, organized by Modern Japan (a magazine): The candidates were Korean women residing in Japan or Korea. The panel of judges evaluated the photographs of the candidates. The victor was a 20-year-old Korean geisha. Her winning photograph was featured in the Korean edition of the magazine Modern Japan. She stood at 157 cm in height and weighed 45 kg (BMI 18.25 kg/m2). As beauty standards evolved over time, the beauty standards of the 1930s were markedly different from those of today.

Aging-Related Changes of the Historical Japanese Beauty, Ono no Komachi (, 825-900 AD).

The aim of this study was to determine how the aging-related changes of a beautiful woman were described in Japanese literature. A poem written by Ono no Komachi was reviewed. Images of her were also searched through Google search. Komachi's poem in Hyakunin Isshu reads: "How the color has faded from the flowers. As I gaze in reverie at the falling rain, I find my beauty, too, has fallen in this world's esteem." Tamatsukuri Komachiko's Death Book describes: "She became thin and looked tired. Her hair looks like a lotus with frost. Her skin resembles a frozen pear. Her bones stick out and her tendons are visible. Her face is dark, and her teeth are yellow". Color woodblock print shows an old woman: She retains her intelligence and beauty even in her advanced years. Her posture was stooped, with a pronounced forward curvature of the spine, resulting as "turtle neck." Her hair was long and gray. Visible signs of aging included forehead wrinkles, crow's feet, deepened nasolabial folds, and marionette lines. Her upper jaw appeared retracted, a "witch's chin." She exhibited neck wrinkles as well as on her hands and wrists, which grasped a bamboo walking stick. Her eyes were sunken, accompanied by a drooping of the upper eyelids. We think that the narrative of her transformation into an ugly and poor woman was intended to convey to men the fleeting nature of women's beauty, and to women that the power of beauty is not ultimately of major significance.

"All Women of Nagoya are Beautiful": Travel Records of a Korean Diplomat in 1764.

The aim of this study is to ascertain how women of that era looked to a Korean diplomat who visited Japan in the 18th century. During 1763-1764, the "Documents on Joseon Tongsinsa (diplomatic mission)" contain 10 records of Korea and 2 records of Japan. Among them, "The Song of a Grand Trip to Japan" (), a Joseon gasa, was reviewed. In the diary entries dated February 2 and 3, 1764, when he was in Nagoya, the author (Kim Ingyeom) described the physical appearance of Japanese women in the city. The peoples of here are bright and beautiful/ All the women are peerless beauties. With eyes like stars and cinnabar-red lips / A clear complexion like white jade, eyebrows like butterflies,/ Hands like budding sprouts, forehead like a cicada. They look like carved ice and as if lumped together with snow. How are they so beautiful despite having the flesh and blood of a human being? Though Zhao Feiyan and Yang Guifei (queens of China) are called the eternal beauties,/ They would look less beautiful compared to women here. If they dressed in Korean costumes and were decorated with jewels,/ We would forget ourselves and mistaken them as goddesses. He highlighted features such as prominent foreheads, eyebrows shaped like butterflies, star-like eyes, red lips, slender hands, and pale skin. He portrayed Japanese women as having a superior appearance compared to Chinese or Korean women. This record suggests that a diplomat admired the appearance of Japanese women during the Meiwa Era (1764-1772).

Aconite-Induced Metamorphosis From Beauty to Ugliness in a Japanese Tale.

The authors present a narrative that details the cause and process of a woman's transformation from beauty to ugliness in a Japanese tale. In "Tokaido Yotsuya Kaidan," the metamorphosis from a beauty to an ugly woman is analyzed. After taking medication to recover from childbirth, Oiwa's face became disfigured and grotesque. Oiwa: It seems to be good for my blood, but when I drink it, it causes fraying and breakdown, causing sudden pain. I feel a numbing dullness. When she looked at herself in the mirror, she was shocked. Oiwa: What's wrong with my face? I hate this, it's such a disgusting thing. Neighbor: It was a lie that the medicine you drank was a helpful medicine that protects the appearance of people's faces. Your face is that of a wicked woman (). Oiwa: Is it really me? Why does she have the face of an evil woman? Her hair falls out in a gruesome combing scene, driving her mad. The poison Oiwa took was aconite, which grows wild in Japan. Both aconite tincture and raw aconite roots contain high concentrations of Aconitum alkaloids, which can penetrate the stratum corneum following the diffusion gradient. As her hair fell out during a horrific combing session, she abandoned her maternal role and sought revenge, having lost what she considered a symbol of her femininity. In treating female patients with facial disfigurement, it is important to be mindful of their psychological state, akin to that of Oiwa, who became disfigured through no fault of her own.

Images of Modern Beauties in the Japanese Colonial Era of Korea (1910-1945).

This study aimed to analyze images of the modern beauty of Korea during the Japanese colonial era. Searches were conducted on Google (www.google.com) and Naver (www.naver.com) for archives of newspapers and magazines that contained images of beauty. Beauty of the face and neck (the early 1920s): an article (1922) detailed the contemporary beauty standards. It specifies the desired characteristics of the eyes, nose, and mouth and dictates that "the cheeks ought to be plump enough to obscure the cheekbones" and "the neck and shoulders should also be full, concealing the collarbones." Images of beauty showing balance and proportion (the late 1920s): in 1928, a magazine article introduced the concept of "the world's beauty from a scientific perspective," which represented Western esthetics as reflected in the "Canon" of body proportions and Vitruvian Man. From the face to the body (from the late 1920s to 1930s): in 1927, a daily newspaper established the standard for global beauty. During this period, there was an increased emphasis on maintaining the body, rather than just the face, as a crucial aspect of beauty. From the mid-1930s, the concept of a "streamlined" female body shape gained popularity. Male gaze, which fragmented and objectified women's bodies, had been visualized and mass-produced. It is essential to understand how preferences have evolved and to possess skilled hands capable of improving the face and body. To properly sculpt the face and body, keen eyes and adept hands are needed.

Lifestyle of Cleft Lip/Palate Patients in Korea Before the Introduction of the Millard Technique.

The authors introduce a short story about the lifestyle of a cleft lip woman in Korea, published in 1936, about 2 decades before Dr Millard operated on his first patient in Korea. A short story entitled "Ugly Thing," written by Yao-hsieh Chu, analyzed by Eonnyeon, a girl born in a rural area with a cleft lip and palate who excelled at household tasks such as sewing. Her forehead was bulging, and her eyes were protruding. She had a low-profile nose with wide nostrils. She had a left unilateral complete primary cleft, including an alveolar cleft. There was midface retraction, accompanied by a relatively prominent chin. Her voice exhibited hypernasality. People often laugh when they see Eonnyeon eating her food, exclaiming, "She's eating like a rabbit!" However, she married a young man who deserted her on their wedding night because of her appearance. Subsequently, she moved to an urban area and found work as a housemaid. One day, she was raped by a middle-aged water delivery man, who then fled. Ten months later, she gave birth to a girl who also had a cleft lip. In a moment of despair, Eonnyeon attempted to smother her baby with a pillow but stopped herself. She resolved to raise her daughter despite the challenges ahead. The mockery directed at her could be viewed as an expression of the latent sadism inherent in human nature. It is remarkable to consider the progress made since Millard performed his first cleft lip surgery in Korea in 1954.

Hand of the 42-armed Avalokitesvara Holding A Skull Rod.

The skull most commonly symbolizes death, mortality, and the unachievable nature of immortality. However, in Korean Buddhist art, skulls are rarely found. The aim of this study was to explore the image and meaning of the skull in the Joseon dynasty (1392 to 1910) of Korea. The terms [Buddhist] and [painting] and [Korea] were used to search Google (www.google.com), and the Research Information Sharing Service of Korea (https://www-riss-kr-ssl.openlink), was also searched for [Buddhism] () and [art] () and [skull] (). This study focused on paintings or statues of Avalokitesvara because Avalokitesvara was often depicted holding various items in the hands, especially the multiarmed Avalokitesvara. The corresponding Buddhist texts were searched on "Archives of Buddhist Culture" (http://kabc.dongguk.edu). In the paintings and statues of the 42-armed Sahasra-bhuja Avalokitesvara, one of the hands was holding a skull rod. In the royal version of the "Five Great Dharanis Collection ()," which was published in 1458, a skull was found in a rod held by a 42-armed Avalokitesvara. This corresponds to one of the verses of the "42-Hand Mantras" (Om dhuna vajra ha). The purpose of the mantra was to communicate with and manipulate ghosts. The meaning in the original Susiddhikara Sutra is quite different from the 42-Hand Mantras in the Joseon dynasty of Korea. The reason for this difference is thought to be the merger of esoteric Buddhism with the folk religions of China and Korea.

Rejuvenation: From the Fictional Dr Heidegger (1837) to Pioneering Modern Plastic Surgeons (1901-1912).

This study aims to identify criticisms of rejuvenation and cosmetic surgery through an analysis of "Dr Heidegger's Experiment" (1837), written by Nathaniel Hawthorne. Dr Heidegger an eccentric, aged scientist, invites 4 elderly friends to be his subjects of study. The guests drink the water from the "fountain of Youth." As their youth is restored, they revert to the foolish behaviors of their younger days. The 3 men soon start vying for the affection of the now youthful and attractive widow. In the midst of their rivalry over the widow, the vase is shattered, and miraculous water spills out. The guests revert to their elderly states. To regain their lost youth, they journey in search of the fountain. Hawthorne viewed rejuvenation procedures as consumptive and physically destructive transformations. His novel captures the modern anxiety surrounding rejuvenation and cosmetic surgery through the lens of literary fantasy. Observing the transformation of the 4 guests, Dr Heidegger understands that youth is too meager a prize to trade for the wisdom that comes with old age. Advancements in medical science have alleviated modern individuals' concerns regarding rejuvenation and cosmetic surgery. Eugen Holländer and Erich Lexer independently performed a facelift in Germany (1901). Charles Conrad Miller first described the blepharoplasty technique in the United States (1906). Suzanne Noël performed a rhytidectomy in France (1912). For the plastic surgeon, it is important to sculpt the face in a way that reflects the patient's virtues, such as wisdom and dignity-qualities that Hawthorne highlighted through the character of Dr Heidegger.

The Psychology of Patients With Oral-Facial Clefts: A Review of Victor Hugo's The Man Who Laughs (L'homme qui rit).

In Victor Hugo's The Man Who Laughs, the protagonist has a facial cleft. The authors aimed to comprehend the psychological experience of a patient with a facial cleft by analyzing this novel. The novel concerns the life of a young nobleman, Gwynplaine, disfigured as a child, who travels with his protector and companion, the vagabond philosopher Ursus, and Dea, a baby girl he rescued. During the oppressive rule of the king, one of his adversaries was Lord Linnaeus Clancharlie, who had taken refuge in Switzerland. The tyrannical monarch ordered the execution of Gwynplaine's father and commissioned a surgeon, Dr Hardquannone, to carve the boy's face into a perpetual grin. The king declared that he should "laugh forever at his fool of a father." Subsequently, the king sold the boy to a group who specialized in the mutilation and disfigurement of children. On the psychology of a boy with an appearance giving the impression of laughing, Hugo wrote: "By laughing Gwynplaine made people laugh. However, he wasn't laughing. His face laughed, his thoughts did not." When Ursus met Gwynplaine, Ursus asked the boy: "What are you laughing at?" The boy replied: I'm not laughing. Ursus: Then you are terrible. Gwynplaine's unnatural laughter, which stems from an artificial deformation of the muscles, is a silent form of laughter. It represents the eternal laughter of those who do not truly laugh, signifying emotional emptiness and an accompanying sense of indifference. Plastic surgeons can restore an "unnatural laughing face" to a "natural smiling face."

Perceptions of Nasal Profile and Augmentation Rhinoplasty in the Japanese Colonial Era of Korea (1910-1945).

The aim of this study was to analyze perceptions of nasal profiles and the origins of augmentation rhinoplasty in Korea during the Japanese colonial era. Through (www.google.com) and (www.naver.com), the archives of newspapers and magazines were searched for references to "nasal augmentation." The following topics were identified: the ideal nasal profile in the corresponding era, nasal augmentation by nonsurgical methods, camouflage makeup for noses with a low profile, the earliest literature on augmentation rhinoplasty in Japan, a case of augmentation rhinoplasty in a Korean woman, an estimate of the year when augmentation rhinoplasty was first performed in Korea, and augmentation rhinoplasty by injection. In 1922, the Nose Lifter (nose uplifting clip) was already commercially available to Korean women. The following year, in 1923, Nishihata and Yoshida published their work on augmentation rhinoplasty using ivory in Japan. By 1925, some hospitals had performed augmentation rhinoplasty, but the procedure had not yet gained popularity among Koreans. In 1927, a Japanese author emphasized the importance of an ideal nasal profile harmonizing with the lips, chin, and neck. Also, in 1927, rhinoplasty gained popularity and was performed at various ENT clinics throughout Korea using various methods. In 1929, a 3-dimensional makeup method was introduced to camouflage a low-profile nose. By 1935, there was a belief that augmentation by injection was not harmful. The authors believe that augmentation rhinoplasty began in 1925 and gained popularity among Koreans by 1927. It was not until 1935 that the complications associated with injectable augmentation were recognized.

Creating Double Eyelids in the Japanese Colonial Era of Korea (1910-1945).

The aim of this study was to analyze the origins of the double-eyelid procedure in Korea during the Japanese colonial era. Through Google (www.google.com) and Naver (www.naver.com), the archives of newspapers and magazines were searched. The following topics were identified: ideal eyes in the Japanese colonial era, creating double eyelids using a non-surgical method, the first case of a double-eyelid operation in a Korean woman, and the presumed first case of a double-eyelid operation in Korea. By 1935, possessing double eyelids had come to be recognized as a symbol of modern beauty. An article suggested that women with small eyes consider surgery to transform their "single eyelid" into a "double-eyelid." It endorsed plastic surgery as a marker of "worldly progress." In 1939, a magazine featured an advertisement for Aihon tape, which was promoted to create "double eyelids." Oh Yeop-ju, the first Korean hairdresser, underwent double-eyelid surgery at an Ophthalmology Clinic in Japan, sometime between 1927 and 1932. She is celebrated as a beauty icon who broadened the beauty horizons of Korean women. After returning in 1933 with a "very beautiful" appearance resulting from successful double-eyelid surgery, she received a special invitation to the Kong Eye Center in Seoul, which opened in 1937. Upon hearing her detailed account and examining her eyelids, the clinic began offering double-eyelid surgery. The technique for double-eyelid surgery was introduced from Japan and disseminated through a medical journal and to patients who had undergone the procedure in Japan.

Features of Ugly but Capable Women of Ancient China.

The aim of this study was to analyze the appearance of women viewed as "ugly but capable" in ancient China. A Google search was conducted for the "Four Great Ugly Women." Information on 3 women was collected from Lienüzhuana, a collection of biographies of eminent women compiled by the Former Han period. With the addition of 1 (Huang Yueying), 7 "ugly but capable" women were analyzed. The manifestations of their ugliness in the texts were recorded. Among the 7 women, 5 exhibited physical manifestations, whereas the remaining 2 were simply described as "ugly" without specific characteristics. Among the discernible body characteristics of the 5 women deemed unattractive, the most common feature was dark skin (80%), followed by body shape (60%). Other frequently observed traits included sparse or yellow hair, an upturned or overly large nose, a convex forehead, and an unattractive neck (each at 40%), with the latter potentially exhibiting a protruding Adam's apple, a thick neck, or a neck mass. Less common features were sunken eyes, a large head, and a masculine skeleton, each occurring in 20% of cases. Based on the depiction of women considered unattractive, the authors can infer that dark skin, a fat body, a large or upturned nose, sparse hair, a protruding forehead, a thick neck, deep-set eyes, and a large head were elements associated with ugliness. Through the description of the ugly woman, authors can infer the standards of ugliness in ancient China.

What Did the Ideal Beauty of the 15th Century Look Like? An Anthropometric Analysis of Botticelli Portraits.

The aim of this study was to conduct an anthropometric analysis of the 5 portraits painted by Botticelli that depict Simonetta Vespucci. Five images in the Simonetta series by Botticelli workshop were measured. The anthropometric measurements of the face included 22 parameters on the lateral view (in 4 portraits; 18 distances and 4 angles) and 17 distances on the frontal view (in one portrait), which were measured using Adobe Photoshop. The absolute distances were calculated relative to the vertical corneal diameter (10.6 mm), which was calculated by multiplying the distance from the pupil's center to the lower margin of the iris. In the lateral faces, the nasofrontal angle (g-n-prn) was 157.6±2.4 degrees, and the nasal tip angle (n-prn-sn) was 99.7±3.4 degrees. The nasolabial angle (prn-sn-ls) was 125.7±4.9 degrees, and the labiomental angle (li-sl-pg) was 131.6±4.4 degrees. The ratio of the upper lip height to the lower lip height (sn-sto/sto-sl) was 85.4±9.0%. The ratio of the upper lip vermillion to the upper lip height (ls-sto/sn-sto) was 27.7±3.9%. The ratio of the lower lip vermillion to the lower lip height (sto-li/sto-sl) was 47.2±6.6%. Comparing the data with 21st-century Italian females, forehead II height (tr-n), physiognomical face height (tr-gn), and morphologic face height (n-gn) of the beauties of the 15th century were significantly greater than those of 21st-century Italian females. However, there were no significant differences in lower face height (sn-gn) and nose height (n-sn). Considering the ongoing cultural relevance of Renaissance art, the esthetic proportions from this study may have reflection to the present day plastic surgery.

Ir(triNHC)-Catalyzed Upcycling of Waste PET for Lactic Acid Production with Sustainable Isolation via Bipolar Membrane Electrodialysis.

For the upcycling of waste polyethylene terephthalate (PET), encompassing both colored and fabric PET materials, we investigated the Ir(triNHC)-catalyzed dehydrogenative coupling of PET and methanol, leading to the production of sodium lactate with good yields. We proposed a sustainable method for isolating lactic acid from the catalytic reaction mixture of sodium lactate and regenerating the base using bipolar membrane electrodialysis (BMED). This isolation method demonstrated high effectiveness, achieving isolation of lactic acid while maintaining economic feasibility at $0.10 per kg of lactic acid, and enabling sustainable NaOH regeneration with complete resource circulation. We assessed the recyclability of the catalyst and elucidated the mechanism involving base-mediated depolymerization and catalyst-promoted dehydrogenation, highlighting the importance of triNHC ligands in enhancing catalytic activity.

Convolutional neural network-based vocal cord tumor classification technique for home-based self-prescreening purpose.

BACKGROUND: In this study, we proposed a deep learning technique that can simultaneously detect suspicious positions of benign vocal cord tumors in laparoscopic images and classify the types of tumors into cysts, granulomas, leukoplakia, nodules and polyps. This technique is useful for simplified home-based self-prescreening purposes to detect the generation of tumors around the vocal cord early in the benign stage. RESULTS: We implemented four convolutional neural network (CNN) models (two Mask R-CNNs, Yolo V4, and a single-shot detector) that were trained, validated and tested using 2183 laryngoscopic images. The experimental results demonstrated that among the four applied models, Yolo V4 showed the highest F1-score for all tumor types (0.7664, cyst; 0.9875, granuloma; 0.8214, leukoplakia; 0.8119, nodule; and 0.8271, polyp). The model with the lowest false-negative rate was different for each tumor type (Yolo V4 for cysts/granulomas and Mask R-CNN for leukoplakia/nodules/polyps). In addition, the embedded-operated Yolo V4 model showed an approximately equivalent F1-score (0.8529) to that of the computer-operated Yolo-4 model (0.8683). CONCLUSIONS: Based on these results, we conclude that the proposed deep-learning-based home screening techniques have the potential to aid in the early detection of tumors around the vocal cord and can improve the long-term survival of patients with vocal cord tumors.

Role of cathepsin D induced by Porphyromonas gingivalis lipopolysaccharide in periodontitis.

Periodontitis is an inflammatory disease of tooth-supporting tissues caused by oral bacteria. Periodontal ligament loss and alveolar bone destruction occur in progressive periodontitis. Since gingival crevicular fluids (GCF) reflects the inflammatory environment of the periodontal pocket, it is a very important specimen for developing targets for periodontitis diagnosis. An antibody array was performed using GCF collected from healthy participants and patients with periodontitis to identify the proteolytic enzymes involved in periodontitis. Of 21 targets on the antibody array membrane, kallikrein 6 (KLK6), kallikrein 10 (KLK10), cathepsin A (CathA), and cathepsin D (CathD) showed higher levels in periodontitis GCF than in GCF from healthy participants. Lipopolysaccharide stimulation of Porphyromonas gingivalis (PG-LPS) in immortalized gingival fibroblasts only increased CathD protein levels among the four targets. The substrate cleavage activity of CathD was increased in PG-LPS-treated immortalized gingival fibroblast extract. The PG-LPS-induced substrate cleavage effect was abolished by the CathD inhibitor pepstatin A. Osteoclast formation was promoted by treatment with conditioned media from PG-LPS- treated immortalized gingival fibroblasts but inhibited by the CathD inhibitor pepstatin A. These results suggest that PG-LPS affected the osteoclast formation process by increasing CathD expression in cells around the alveolar bone, thereby participating in periodontitis progression.