Diverse effects of clarithromycin and proposal of its clinical application for treating COVID-19 as a repurposing drug.

Outbreak of SARS-CoV-2 has been declared a pandemic, which is a serious threat to human health. The disease was named coronavirus disease 2019 (COVID-19). Until now, several vaccines and a few drugs have been approved for the prevention and treatment for COVID-19. Recently, the effect of some macrolides including clarithromycin (CAM) on COVID-19 has attracted attention. CAM is known to have diverse effects including immunomodulatory and immunosuppressive effects, autophagy inhibition, steroid sparing effect, reversibility of drug resistance, antineoplastic effect, antiviral effect as well as bacteriostatic/bactericidal effect. Many patients with COVID-19 died due to an overwhelming response of their own immune system characterized by the uncontrolled release of circulating inflammatory cytokines (cytokine release syndrome [CRS]). This CRS plays a major role in progressing pneumonia to acute respiratory distress syndrome (ARDS) in COVID-19 patients. It is noteworthy that CAM can suppress inflammatory cytokines responsible for CRS and also has anti-SARS-CoV-2 effect. Considering the rapidly progressive global disease burden of COVID 19, the application of CAM for treating COVID-19 needs to be urgently evaluated. Recently, an open-labeled non-randomized trial using CAM for treating COVID-19 (ACHIEVE) was initiated in Greece in May, 2020. Its results, though preprint, indicated that CAM treatment of patients with moderate COVID-19 was associated with early clinical improvement and containment of viral load. Thus, treatment with CAM as a single agent or combined with other anti-SARS CoV-2 drugs should be tried for treating COVID-19. In this article, we discussed the significance and usefulness of CAM in treating COVID-19.

Discovery of cockroach specimens of the genus Squamoptera (Ectobiidae: Pseudophyllodromiinae) from Okinawa, Japan, showing wing polymorphism.

Cockroach specimens of the genus, Squamoptera were collected from the Iriomote island of Okinawa prefecture, Japan. The morphological features of the specimens were characterized as having a white band on the dorsal surface of its thorax, its tegmen reduced into a tiny scale-like structure and the hindwing was absent. Ocelli was also absent and the small compound eyes not extending to apex of the head nor to the frontal face but extend further lower than the base of the antennae. When the specimens were reared in the laboratory, besides the short wing form, the long wing form began to appear in the rearing colony. In our reproductive biological study, we observed that hatching of the ootheca from the short wing female takes about 30 days, with an average of 6.6 nymphs being hatched from one ootheca. The male to female ratio of the offspring was 36:30. However, the frequency appearance of the offspring from the ootheca of the short wing female was 98.5% short wing and 1.5% long wing form. Our specimens occasionally show body polymorphism in the form of individuals having long wings instead of the usual short one. The long wing form does not show the white band on the dorsal surface of its thorax.

A new species of cockroach, Periplaneta gajajimana sp. nov., collected in Gajajima, Kagoshima Prefecture, Japan.

We described a new species of cockroach, Periplaneta gajajimana sp. nov., which was collected in Gajajima, Kagoshima-gun Toshimamura, Kagoshima Prefecture, Japan, on November 2012. The new species is characterized by its reddish brown to blackish brown body, smooth surface pronotum, well developed compound eyes, dark brown head apex, dark reddish brown front face and small white ocelli connected to the antennal sockets. In male, the tegmen tip reach the abdomen end or are slightly shorter, while in the female, it does not reach the abdominal end and exposes the abdomen beyond the 7th abdominal plate. We confirmed the validity of this new species by breeding the specimens in our laboratory to demonstrate that the features of the progeny were maintained for several generations. For comparison and easy identification of this new species, the key to species identification of the genus Periplaneta that had been reported in Japan to date are also presented.

Serum liver enzyme profile in Timor deer (Cervus timorensis) with fascioliasis in Indonesia.

Timor deer (Cervus timorensis) at Surabaya zoo, Indonesia, that were found to be naturally infected with Fasciola, showed elevated level of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP). Of a total of 75 deer examined, 12 (25%) of the 47 adult deer and 8 (29%) of the 28 juvenile deer were found to be infected with fascioliasis, as evidenced by the shedding of the parasite eggs. The level of ALT, AST and ALP were significantly elevated (p<0.05) in all the infected deer. Only Fasciolainfected deer showed elevated serum liver enzyme. Deer with elevated enzyme level show a trend that positively correspond with higher Egg per gram of feces (EPG). The average size of the parasite eggs at 169.0±11.1 × 96.0±3.5µm, correspond well with that of Fasciola gigantica. No other trematode eggs were observed besides that of F. gigantica. There was no significant difference in the enzyme profile between the two sexes in both the infected and the uninfected group. This is the first report of the elevation of serum liver enzyme in Timor deer that is associated with not only fascioliasis and also correspond positively with the EPG.

Diverse effects of clarithromycin and proposal of its clinical application for treating COVID-19 as a repurposing drug

@#Outbreak of SARS-CoV-2 has been declared a pandemic, which is a serious threat to human health. The disease was named coronavirus disease 2019 (COVID-19). Until now, several vaccines and a few drugs have been approved for the prevention and treatment for COVID-19. Recently, the effect of some macrolides including clarithromycin (CAM) on COVID-19 has attracted attention. CAM is known to have diverse effects including immunomodulatory and immunosuppressive effects, autophagy inhibition, steroid sparing effect, reversibility of drug resistance, antineoplastic effect, antiviral effect as well as bacteriostatic/bactericidal effect. Many patients with COVID-19 died due to an overwhelming response of their own immune system characterized by the uncontrolled release of circulating inflammatory cytokines (cytokine release syndrome [CRS]). This CRS plays a major role in progressing pneumonia to acute respiratory distress syndrome (ARDS) in COVID-19 patients. It is noteworthy that CAM can suppress inflammatory cytokines responsible for CRS and also has anti-SARS-CoV-2 effect. Considering the rapidly progressive global disease burden of COVID- 19, the application of CAM for treating COVID-19 needs to be urgently evaluated. Recently, an open-labeled non-randomized trial using CAM for treating COVID-19 (ACHIEVE) was initiated in Greece in May, 2020. Its results, though preprint, indicated that CAM treatment of patients with moderate COVID-19 was associated with early clinical improvement and containment of viral load. Thus, treatment with CAM as a single agent or combined with other anti-SARSCoV- 2 drugs should be tried for treating COVID-19. In this article, we discussed the significance and usefulness of CAM in treating COVID-19.

Discovery of cockroach specimens of the genus Squamoptera (Ectobiidae: Pseudophyllodromiinae) from Okinawa, Japan, showing wing polymorphism

@# Cockroach specimens of the genus, Squamoptera were collected from the Iriomote island of Okinawa prefecture, Japan. The morphological features of the specimens were characterized as having a white band on the dorsal surface of its thorax, its tegmen reduced into a tiny scale-like structure and the hindwing was absent. Ocelli was also absent and the small compound eyes not extending to apex of the head nor to the frontal face but extend further lower than the base of the antennae. When the specimens were reared in the laboratory, besides the short wing form, the long wing form began to appear in the rearing colony. In our reproductive biological study, we observed that hatching of the ootheca from the short wing female takes about 30 days, with an average of 6.6 nymphs being hatched from one ootheca. The male to female ratio of the offspring was 36:30. However, the frequency appearance of the offspring from the ootheca of the short wing female was 98.5% short wing and 1.5% long wing form. Our specimens occasionally show body polymorphism in the form of individuals having long wings instead of the usual short one. The long wing form does not show the white band on the dorsal surface of its thorax.

A new species of cockroach, Periplaneta gajajimana sp. nov., collected in Gajajima, Kagoshima Prefecture, Japan

@#We described a new species of cockroach, Periplaneta gajajimana sp. nov., which was collected in Gajajima, Kagoshima-gun Toshimamura, Kagoshima Prefecture, Japan, on November 2012. The new species is characterized by its reddish brown to blackish brown body, smooth surface pronotum, well developed compound eyes, dark brown head apex, dark reddish brown front face and small white ocelli connected to the antennal sockets. In male, the tegmen tip reach the abdomen end or are slightly shorter, while in the female, it does not reach the abdominal end and exposes the abdomen beyond the 7th abdominal plate. We confirmed the validity of this new species by breeding the specimens in our laboratory to demonstrate that the features of the progeny were maintained for several generations. For comparison and easy identification of this new species, the key to species identification of the genus Periplaneta that had been reported in Japan to date are also presented.

Serum liver enzyme profile in Timor deer (Cervus timorensis) with fascioliasis in Indonesia

@#Timor deer (Cervus timorensis) at Surabaya zoo, Indonesia, that were found to be naturally infected with Fasciola, showed elevated level of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP). Of a total of 75 deer examined, 12 (25%) of the 47 adult deer and 8 (29%) of the 28 juvenile deer were found to be infected with fascioliasis, as evidenced by the shedding of the parasite eggs. The level of ALT, AST and ALP were significantly elevated (p<0.05) in all the infected deer. Only Fasciolainfected deer showed elevated serum liver enzyme. Deer with elevated enzyme level show a trend that positively correspond with higher Egg per gram of feces (EPG). The average size of the parasite eggs at 169.0±11.1 × 96.0±3.5μm, correspond well with that of Fasciola gigantica. No other trematode eggs were observed besides that of F. gigantica. There was no significant difference in the enzyme profile between the two sexes in both the infected and the uninfected group. This is the first report of the elevation of serum liver enzyme in Timor deer that is associated with not only fascioliasis and also correspond positively with the EPG.

Experimental Infection with Baylisascaris Potosis in Chickens.

The larvae of the genus Baylisascaris can cause larva migrans in mammals and birds. This study investigated the larval migration of Baylisascaris potosis, the roundworm of kinkajou (Potos flavus), in chickens and the associated clinical manifestations of the host. Thirty-six 3-week-old chickens divided into 6 groups were orally inoculated with 3,000 B. potosis eggs/chick. Each group of chicken was necropsied at days 1, 2, 3, 7, 30 and 90 PI (post inoculation), and the number of larvae in various organs were counted until day 90 PI. No clinical signs were observed in chickens during the study. Larvae were detected from the liver, lungs or breast-muscles of 13/36 (36.1%) chickens. The mean total number of larvae in the liver, lungs and breast-muscles at days 1, 2, 3, 7, 30 and 90 PI were 0.34, 0.17, 1.66, 1.01, 0.17 and 0, respectively. No larvae were found in the brain, eyes, hid-limb muscles, heart, kidneys and spleen. Although infectivity of larvae in egg-inoculated chickens was low, the present study demonstrated that B. potosis larvae can migrate in chickens tissues up to day 30 PI. The result suggests that chickens can serve as a paratenic host for B. potosis and may underline a public health importance of B. potosis infection as a potential foodborne disease in humans.

Baylisascaris Potosis Larvae in Mice of Different Strains and Infectivity of Tissue Larvae.

Migration of Baylisascaris potosis larvae in different mouse strains were compared, and infectivity of the persisting larvae in mice tissues were investigated. Five strains of mice, BALB/c, C57BL/6, AKR, B10.BR, and ICR were inoculated with 1,000 B. potosis eggs/mouse, and necropsied at week 13 post inoculation (PI). The other uninfected ICR mice (secondary host) were inoculated with 43 larvae/ mouse recovered from mice at week 13 PI with eggs, and necropsied at day 21 PI. Larvae in organs or tissues were counted at necropsy. One AKR mouse showed torticollis and circling at day 56 PI. At necropsy at week 13 PI, larvae were recovered from all mice. A mean total larvae recovered were 124.1 (n=40). Majority of larvae were found in the carcass (mean 113.9) and some in the viscera (mean 9.9). Zero to 1 larva were found in the brain or eyes of some mice. There were no differences among the mouse strains in the number of larvae, except in the viscera; more larvae were seen in BALB/c or ICR than in B10.BR mice. No larvae were found in the secondary host mice. Present study demonstrated that B. potosis larvae migrate well in the carcass of any strains of mice, however, the tissue larvae did not infect the secondary host. Results of our present study suggest that B. potosis larvae is less aggressive for the nervous tissue migration than that of B. procyonis larvae which is commonly known to migrate in central nervous system of mammals and birds.

Species clarification of Ogasawara cockroaches which inhabit Japan.

The so-called "Ogasawara cockroaches" were examined by morphological observations and by breeding experiments to elucidate their actual taxonomical status. Fourteen groups (isolate) of "Ogasawara cockroaches" collected from Iwoto-A, Iwoto-B, Hahajima, Chichijima, Nishijima, Nakodojima, Tokunoshima-A, Tokunoshima-B, Okinawato- A, Okinawa-B, Amamiooshima, Miyakojima, Ishigakijima and Hawaii, were bred and passaged in our laboratory. Cockroaches collected from the field were first reared individually and the sexes of their offspring examined. Cockroaches collected from Iwoto, Tokushima and Okinawa, were found to consist of two groups; those whose offspring were all female and the other whose offspring consist of both male and female. Cross-breeding experiments showed that individuals from the group that did not produce any male but only female offspring were parthenogenetic. On the contrary, the group that have bisexual individuals produced both male and female offspring in a ratio of 1:1. Our results show that the so-called "Ogasawara cockroaches" consist of 2 species, namely, Pycnoscelus surinamensis and Pycnoscelus indicus. There are areas in which both species co-habitated together and there are also areas in which either only one of the two species can be found. The group that reproduces only female offspring and only through parthenogenesis was identified as P. surinamensis. The group that reproduces heterosexually and produce male and female offspring was identified as P. indicus. Thus, the so-called "Ogasawara cockroaches" found in Japan actually consist of 2 species, namely, P. surinamensis and P. indicus, which can be differentiated using the solitary breeding method to demonstrate parthenogenesis in the former and the need for sexual reproduction in the latter.

DNA polymerase hybrids derived from the family-B enzymes of Pyrococcus furiosus and Thermococcus kodakarensis: improving performance in the polymerase chain reaction.

The polymerase chain reaction (PCR) is widely applied across the biosciences, with archaeal Family-B DNA polymerases being preferred, due to their high thermostability and fidelity. The enzyme from Pyrococcus furiosus (Pfu-Pol) is more frequently used than the similar protein from Thermococcus kodakarensis (Tkod-Pol), despite the latter having better PCR performance. Here the two polymerases have been comprehensively compared, confirming that Tkod-Pol: (1) extends primer-templates more rapidly; (2) has higher processivity; (3) demonstrates superior performance in normal and real time PCR. However, Tkod-Pol is less thermostable than Pfu-Pol and both enzymes have equal fidelities. To understand the favorable properties of Tkod-Pol, hybrid proteins have been prepared. Single, double and triple mutations were used to site arginines, present at the "forked-point" (the junction of the exonuclease and polymerase channels) of Tkod-Pol, at the corresponding locations in Pfu-Pol, slightly improving PCR performance. The Pfu-Pol thumb domain, responsible for double-stranded DNA binding, has been entirely replaced with that from Tkod-Pol, again giving better PCR properties. Combining the "forked-point" and thumb swap mutations resulted in a marked increase in PCR capability, maintenance of high fidelity and retention of the superior thermostability associated with Pfu-Pol. However, even the arginine/thumb swap mutant falls short of Tkod-Pol in PCR, suggesting further improvement within the Pfu-Pol framework is attainable. The significance of this work is the observation that improvements in PCR performance are easily attainable by blending elements from closely related archaeal polymerases, an approach that may, in future, be extended by using more polymerases from these organisms.

Paclitaxel plus Carboplatin Chemotherapy for Primary Peritoneal Carcinoma: A Study of 22 Cases and Comparison with Stage III-IV Ovarian Serous Carcinoma.

The aim of this study was to assess the clinical characteristics and outcome of patients with either primary peritoneal carcinoma (PPC) or ovarian serous carcinoma (OSC) treated with paclitaxel plus carboplatin chemotherapy. We retrospectively identified 22 PPC patients and 55 stage III-IV OSC patients treated between 2002 and 2007. After exploratory laparotomy, all patients received paclitaxel and carboplatin every 3 weeks, with the goal of optimal cytoreduction. There were no statistically significant differences between the PPC and OSC groups with regard to tumor stage, residual tumor after debulking surgery (initial or interval), serum cancer antigen (CA) 125 levels at diagnosis, and completion of first-line chemotherapy. The progression-free survival (PFS) durations were 12.7 months (95% CI, 6.3-18.5) in the patients with PPC and 15.9 months (95% CI, 13.3-18.5) in those with OSC (p = 0.016). However, the median survival durations were 26.5 months (95% CI, 14.6-38.3) in the patients with PPC and 38 months (95% CI, 23.8-53.8) in those with OSC (p = 0.188). Survival was longer for all patients whose CA125 levels normalized to 26 U/ml during and after treatment. Overall survival (OS) of the patients with PPC was similar to that of the patients with OSC, suggesting that management for advanced-stage OSC would be similar to that for PPC. The combination of optimal debulking with paclitaxel plus carboplatin chemotherapy may offer patients the most effective treatment. The CA125 nadir after cytoreductive surgery can be considered a prognostic factor for OS and PFS in patients with PPC.

Insulin pump therapy in children and adolescents with Type 1 Diabetes: improvements in glycemic control and patients' satisfaction -- Hospital UKM experience.

Hospital UKM (Universiti Kebangsaan Malaysia) introduced the use of insulin pump therapy in children and adolescents with Type 1 Diabetes in Malaysia in April 2004. This study aims to evaluate the effectiveness of pump therapy and its impact on metabolic control among patients from our institution. Insulin pump therapy resulted in sustainable improvement in glycemic control throughout the six years of treatment with reduction in HbA1c in the first two years of pump use was statistically significant. The BMI SDS showed an increase trend but the changes before and after pump use was insignificant. There is also high level of treatment satisfaction reported among our insulin pump patients.

Serum CA125 level before the development of ovarian cancer.

BACKGROUND: Little is known about the natural history of ovarian cancer with respect to the change of serum CA125 level. METHODS: The Shizuoka Cohort Study on Ovarian Cancer Screening (SCSOCS) Trial contains approximately 100,000 data on serum tumor marker CA125 prospectively obtained from more than 70,000 women. We reviewed the clinical charts and collected serum samples 2 months to 9.4 years prior to the surgery were available. RESULTS: In 396 (95%) of the 419 patients with ovarian cancer, one serum sample was present before the diagnosis (mean, 4.1 years). The change of CA125 level before the diagnosis of ovarian cancer could be clearly separated into two groups according to the length of the following intervals: 47% (107/228) of patients with non-serous-type ovarian cancers develop secondarily from slightly elevated CA125 level (35

Congenital rubella syndrome with positive serology and virus isolation.

An effective live attenuated rubella vaccine was available since 1969 and congenital rubella syndrome can be prevented with appropriate vaccination. We report a baby with congenital rubella syndrome born in Klang valley to indicate that the Universal Rubella Vaccination Programme adopted by the Ministry of Health Malaysia since 2002 has yet to achieve its effect of eliminating transmission of rubella and preventing congenital rubella infection in the community. To our knowledge, the virus isolate represents the first successful isolation of rubella virus in this country and will serve as the reference strain for future comparison in molecular epidemiological tracking of rubella virus activity this country.

Development of Taenia asiatica cysticerci to infective stage and adult stage in Mongolian gerbils.

The development of metacestodes and adult worms of Taenia asiatica in Mongolian gerbils (Meriones unguiculatus) were observed. Cysticerci were recovered from gerbils subcutaneously injected with hatched oncospheres. The recovery rate ranged from 0.1 to 3.2%. No cysticerci were recovered from the orally inoculated gerbils. The infectivity of the cysticerci recovered at 48 weeks post-infection was evaluated. Tapeworms were recovered on day 14 post-infection from the small intestine of 5 of 11 gerbils, with a recovery rate of 27% (6 worms recovered/22 worms inoculated). Three and four adult worms were recovered from two human volunteers who ingested five cysticerci after 4 months post-infection. In worms recovered from gerbils, segmentation and genital primordia in the posterior proglottids and hooklets in the residual rostellum were observed. The results indicate that gerbils can serve as an alternative intermediate host and that partial development of the adult worm stage occurs in gerbils.

Development of Taenia saginata asiatica metacestodes in SCID mice and its infectivity in human and alternative definitive hosts.

Development of Taenia saginata asiatica metacestodes in SCID mice, and its infectivity in humans, golden hamsters, and Mongolian gerbils as alternative definitive hosts, were investigated. Cysticerci were recovered from SCID mice that were subcutaneously injected with hatched oncospheres of T. s. asiatica. The morphological changes of metacestodes were observed. The recovered cysticerci were fed to gerbils, hamsters and humans, to check for their infectivity. Tapeworms were recovered from gerbils and hamsters fed with 20 to 45 week-old cysticerci, and proglottids excretions were observed in human volunteers fed with 45 week-old cysticerci. However, no tapeworms were recovered from gerbils fed with 10 week-old cysticerci. Our results suggest that T. s. asiatica oncospheres needed more than 20 weeks to develop to maturity in SCID mice to be infective to both their natural and alternative definitive hosts.

Retarded gastric acid secretion in rats infected with larval Taenia taeniaeformis.

The influence of hepatic larval Taenia taeniaeformis infection on gastric acid secretory activity and gastric mucosal integrity was investigated. After 12 weeks of infection with 2,000 T. taeniaeformis eggs, the gastric pH values of control and infected rats were 4.1+/-0.6 (mean +/- SD) and 8.4+/-0.2, respectively. There was no difference in the basal acid secretion between control (1.7+/-0.7 micro Eq.H(+)/15 min) and infected (1.9+/-0.3) rats. However, infected rats failed to respond to histamine stimulation, the maximum acid output level being 2.8+/-0.4 in the infected rats, compared to 12.9+/-3.3 in control rats. Larval T. taeniaeformis infection resulted in the suppression of gastric acid secretion leading to hypergastrinemia.