Efficacy of imidacloprid/permethrin and fipronil/(S)-methoprene combinations against Haemaphysalis longicornis ticks evaluated under in vitro and in vivo conditions.

Haemaphysalis longicornis is one of the most important ticks infesting a wide range of mammals including dogs in Japan. H. longicornis is recorded to be a vector of, for example, Babesia gibsoni. It was the aim of the study presented here to evaluate the efficacy of imidacloprid/permethrin and fipronil/(S)-methoprene against larval, nymphal and adult stages of H. longicornis under in vitro as well as in vivo conditions. In the in vitro part of the study, ticks showed avoidance behaviour to imidacloprid/permethrin-treated filter papers. The onset of acaricidal efficacy in the imidacloprid/permethrin group was recorded earlier than in the fipronil/(S)-methoprene group. In the in vivo experiment three beagles per group were treated with either imidacloprid/permethrin, fipronil/(S)-methoprene or left untreated. Each dog was infested with 30 adult female H. longicornis. Ticks were place on a shaved area of skin of the treated dogs and behaviour of the ticks was recorded as before. After 3 h all ticks were removed and placed in Petri dishes. Ticks were further examined until day 4 post-treatment (p.t.). All ticks recovered from the untreated dogs survived. At 4 h p.t. (1 h post-removal) 40 of the 90 ticks exposed to the imidacloprid/permethrin treatment and 25 of the 90 ticks in the fipronil/(S)-methoprene-treated group were found dead. At day 1 p.t., 61 ticks in the imidacloprid/permethrin- and 81 ticks in the fipronil/(S)-methoprene-treated group were recorded dead. At the final examination day 4 p.t., all 90 ticks were found dead in the imidacloprid/permethrin group, while five ticks remained alive in the fipronil/(S)-methoprene group.

Two cases of Hashimoto's thyroiditis with transient hypothyroidism.

Two cases of Hashimoto's thyroiditis are presented: a woman who suffered twice from transient hypothyroidism (Case 1), and a woman with polycystic ovary syndrome who had transient hypothyroidism which was inferred to have been caused by exacerbation of Hashimoto's disease (Case 2). In both cases, fluctuation in titers of both anti-thyroglobulin (TGHA) and antimicrosomal antibodies (MCHA) was observed. Although an increased serum thyroid stimulating hormone (TSH) concentration in Case 2 was associated with the increased titer of MCHA, this was not true of Case 1. Measurement of serum iodine concentration in Case 1 disclosed no correlation between serum TSH and iodine concentrations, suggesting that episodes of hypothyroidism in this patient are not due to iodine-induced hypothyroidism. The transient hypothyroidism in Case 2 was considered to be due to fluctuations in immune mechanism(s), but the reason in Case 1 was not clear in the present study.

Inactive renin release from the human arm vascular wall during upper arm occlusion: relation to beta-adrenoceptors.

To investigate the extrarenal release of renin, plasma active renin (PAR) and plasma trypsin activatable or inactive renin (PIR) in blood taken simultaneously from bilateral median cubital veins were measured in seven normal men before and after occlusion of the right upper arm with a standard cuff inflated halfway between systolic and diastolic blood pressures for 15 min. After 15 min, both PAR (P less than 0.05) with PIR (P less than 0.01) increased significantly in occluded arms compared with 0 min. PIR rose significantly (P less than 0.01) in occluded arms compared with controls at 15 min, but PAR was unchanged. The same studies were repeated after pretreatment with oral doses of either 1 mg prazosin or 10 mg propranolol at 1600 h and 2400 h on the previous day and 1 hour before the experiment. After prazosin, both PAR (P less than 0.05) and PIR (P less than 0.01) in occluded arms increased significantly at 15 min compared with those at 0 min and PIR rose significantly (P less than 0.01) in occluded arms compared with controls at 15 min, as observed in the control studies. On the other hand, after propranolol, the significant increment of PAR in the occluded arms was abolished. Moreover, the significant difference of PIR between the occluded and the control arms at 15 min was also abolished. These results indicate that inactive renin may be present in the vascular wall of the arm and released into circulation by a stimulus such as arm occlusion by mechanisms related to beta-adrenoceptors.

[Release mechanisms of inactive renin from rat renal cortical slices: role of the submandibular gland].

In our previous studies, we showed an in vivo stimulating effect of the extract of the rat submandibular gland on plasma inactive renin release. In this study, we evaluated the effects of the rat submandibular gland extract and of some plasma active renin stimulants on inactive renin release from rat renal cortical slices. Adult male Wistar rats (250-350g) were kept on a regular diet (Na 260mg/100g) and nephrectomized under pentobarbital anesthesia (50mg/kg, i.p.). Five thin renal cortical slices were obtained from each kidney by using a razor blade. These renal cortical slices were incubated in Earle's buffer (pH7.4, Difco) at 37 degrees C for 30 min (preincubation), then transferred into 10ml fresh Earle's buffer with or without some agents and incubated at 37 degrees C for 1 hour (experimental incubation). For each experiment, 6 groups of 5 renal cortical slices were employed. The agents used in this study were as follows: isoproterenol (10(-5)M), furosemide (50 micrograms/ml), prostaglandin E1 (10(-5)M), prostaglandin I2 (10(-5)M) and the rat submandibular gland extract (100 microliters) which was obtained after homogenation with 10 x (w/v) 0.01M pyrophosphate buffer (pH6.5) including 0.1M NaCl. One ml of samples of this Earle's buffer were withdrawn every 20 min. Active renin in the samples was assayed by the commercial RIA-kit (Dainabot), and total renin was assayed after trypsin (Worthington) treatment (30 micrograms/300 microliters sample) at 4 degrees C for 10 min. Inactive renin was determined as the difference between total renin and active renin. Active and inactive renins increased linearly in the buffer without any agents (control) during the observation period (60 min). Isoproterenol (10(-5)M) stimulated the release of active renin significantly (p less than 0.01 vs. control) but did not affect the release of inactive renin. Furosemide (50 micrograms/ml) stimulated the release of active and inactive renins significantly at 20 and 40 min (p less than 0.05 vs. control) but did not affect the release of either renin at 60 min. Both prostaglandins E1 and I2 (10(-5)M) stimulated the release of active renin significantly (p less than 0.01 vs. control) but inhibited, on the other hand, the release of inactive renin significantly (p less than 0.01 vs. control). The rat submandibular gland extract (100 microliters) did not affect the release of active renin but stimulated the release of inactive renin significantly (p less than 0.05 vs. control).(ABSTRACT TRUNCATED AT 400 WORDS)

[Roles of the kidney in activation and release of plasma inactive renin in the rat].

Plasma inactive as well as active renin is supposed to originate from the kidney, though there is little direct evidence. As we have previously reported (Sakanaka et al., Folia Endocrinol. Jap., 63: 961-977, 1987; Miyazaki et al., J. Hypertension 4 (Suppl 6): S453-S455, 1986; Miyazaki et al., J. Hypertension 6: 33-40, 1988), the submandibular gland, but not the kidney, is thought to play an crucial role in releasing plasma inactive renin in the rat. In the present acute studies, we attempted to elucidate the roles of the kidney in the release mechanisms of plasma inactive renin. Adult male rats maintained on a regular rat chow (Na: 260 mg/100g) were uninephrectomized, and vessel clips were placed on the renal pedicles of the contralateral kidneys to make completely ischemic and non-filtered kidneys. In the first protocol, the renal pedicles were occluded for 2 h, followed by the removal of the vessel clips. During the occlusion for 2 h, plasma active renin concentration (PAC) in the peripheral blood obtained from the femoral cannulae decreased, while plasma inactive renin concentration (PIC) along with plasma total renin concentration (PTC) increased as in the case of total nephrectomy, which supports our previous studies. Then, declipping resulted in the rapid rise in PAC with the peak values at 2 min, which was followed by its gradual decrease with time during the experimental period (30 min). On the other hand, PIC decreased gradually toward control levels with no rise after declipping. In the second protocol, blood trapped in the kidney was collected through the renal venous cannulae at 0, 60, 120 and 240 min after the pedicle occlusion in the different groups of rats. The renal blood levels of PAC increased by more than three times at 240 min compared to the control values, while PIC decreased to one third of the control values. PTC increased at 120 and 240 min. Renal tissue levels of renin were also measured at 0 and 120 min in the second protocol in the kidneys of rats which were maintained on a regular rat chow. Inactive renin concentration increased, while active renin concentration decreased. These were compatible with the results obtained in plasma. In the last protocol, the second protocol was in part repeated in salt-depleted rats which were kept on a low salt diet (Na: 11.3 mg/100g) for 2 weeks.(ABSTRACT TRUNCATED AT 400 WORDS)

[Role of the submandibular glands in in vivo mechanisms of plasma inactive renin release in the rat].

In our present studies, we evaluated the role of the submandibular glands (SMG) on plasma inactive renin (PIR) releasing mechanisms in rats using some agents which are known to stimulate plasma active renin (PAR) release. The results were analyzed between sialoadenectomized (SX) and sham-operated (control: C) rats. Twenty-four h after the operation, PAR releasing agents, furosemide (FRO) 2.5 mg/rat/h with prior iv bolus 5 mg, captopril (CAP) 5 mg/rat/h with prior iv bolus 10 mg, 1-Sar-8-Ile-angiotensin II (Ang II A) 300 ng/kg/min, prostaglandin E1 (PGE1) 100 ng/kg/min, and prostaglandin I2 (PGI2) 100 ng/kg/min, were infused through femoral venous cannulae. Blood samples were taken through femoral arterial cannulae into test tubes containing 2 mg EDTA-2Na. PAR was assayed by RIA, and total renin was obtained after tryptic activation. According to the responses of PIR, the agents used were categorized into three patterns: FRO increased PIR, both PGs lowered PIR, and, CAP and Ang II A had no effect on PIR release. The PIR release mechanisms by FRO were further investigated by 20 mg FRO ip injection in totally nephrectomized rats. PIR increased even in nephrectomized rats, but the increase was totally canceled by the following SX. In conclusion, FRO alone among some agents studied is able to stimulate PIR release only under the existence of SMG.

Increased furosemide-sensitive sodium influx into erythrocytes in non-insulin-dependent diabetes mellitus associated with hypertension.

Our previous study demonstrated increased sodium (Na+) influx into ouabain-treated erythrocytes in diabetic patients with hypertension. The present study was designed to estimate the effect of furosemide on the Na+ influx in non-insulin-dependent diabetes mellitus (NIDDM) with hypertension. Both Na+ influx in the absence of furosemide (total Na+ influx; TNaI) and furosemide-sensitive Na+ influx (FSNaI) were increased in patients with essential hypertension (group 1, n = 22) than those in non-diabetic, normotensive control subjects without family history of hypertension (control group, n = 26, P less than 0.001 and P less than 0.001, respectively). TNaI and FSNaI in diabetic patients without hypertension (group 2, n = 28) were also higher than those in the control group (P less than 0.02 and P less than 0.001, respectively), but no significant differences were found in TNaI or FSNaI between groups 1 and 2. TNaI and FSNaI in diabetic patients with hypertension (group 3, n = 19) were elevated than those in control group (P less than 0.001 and P less than 0.001, respectively), in group 1 (P less than 0.05 and P less than 0.02, respectively) and in group 2 (P less than 0.001 and P less than 0.001, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Reno-submandibular axis controls release of extrarenal inactive renin.

The mechanisms causing the release of plasma inactive renin (PIR) area still unclear. We have investigated the role of the kidney in the release of trypsin-activable PIR from extrarenal sources in the rat, with special reference to the submandibular gland. The activation of PIR was performed by incubation with 20 mg/ml trypsin at 4 degrees C for up to 10 min; the reaction was then terminated by addition of 20 mg/ml of soybean trypsin inhibitor. Bilateral nephrectomy resulted in a gradual, marked, sex-independent increase in PIR concentration, reaching levels 4.5 times higher than basal in 24 h (time 0: 14.8 +/- 1.0 ng/ml per h; 24 h: 66.8 +/- 3.4 ng/ml per h, mean +/- s.d., P less than 0.001). This increase was not altered by the concomitant intravenous infusion of pressor doses of either angiotensin (Ang) II (30 ng/min) or pure mouse submandibular renin (a 20-ng intravenous bolus followed by intravenous infusion at the rate of 50 ng/h) for 4 h, but was completely prevented by prior removal of the submandibular glands, in which no activity of active renin and no inactive renin was detected. These results suggest that the post-nephrectomy increase of PIR is not dependent on feedback mechanisms of the suppressed renin-angiotensin system, but is controlled by the presence of submandibular glands in the rat.

[Roles of kidney and submandibular gland in the release of rat plasma inactive renin].

In this study we outlined the development of an enzymatic technique to activate plasma inactive renin by trypsin in rat plasma. Using this method, we reported the releasing mechanism of the trypsin-activable inactive renin which has not yet been clarified. Adult male Wistar rats (260-300 g) were kept on regular diet (Na: 260 mg/100g) unless explained and underwent operation under pentobarbital anesthesia (50 mg/kg). Blood samples were obtained from conscious rats through the cannulae, which had been inserted into the left femoral arteries 24h before the experiments. After addition of excessive renin substrate which had been obtained from the 24 h-nephrectomized rat plasma, renin was measured by the commercial RIA-kit (Dainabot). Trypsin (Worthington) treatment (20 mg/ml plasma for 10 min at 4 degrees C) was followed by addition of SBTI (Sigma) (20 mg/ml plasma). This condition maximally increased the rate of angiotensin I generation and did not alter the Km or optimum pH of the renin reaction. In this condition, trypsin reaction was completely inhibited by adding these concentrations of SBTI. The molecular weight of inactive renin (51,000) in the normal rat plasma estimated by Sephadex G-100 column (Pharmacia) was the same as that in the nephrectomized rat plasma. In conclusion, trypsin treatment of plasma (20 mg/ml plasma for 10 min at 4 degrees C) followed by SBTI (20 mg/ml plasma) was justified for trypsin activation of rat plasma. Using this method, we investigated the changes in active and inactive renin after bilateral nephrectomy in the salt-depleted rat. Active renin decreased rapidly after bilateral nephrectomy with a half life of 23.6 +/- 4.0 min. Inactive renin, on the other hand, increased gradually and reached to a plateau 24 h after bilateral nephrectomy, and was kept unchanged during the following 24 h. The infusion of mouse submandibular gland active renin or angiotensin II could not prevent the increase of plasma inactive renin in the nephrectomized rat. These suggest that there may be no feedback mechanisms between plasma inactive and active renin or angiotensin II. Furthermore, we investigated the organ-related sources of plasma inactive renin which markedly increased after total nephrectomy. Simultaneous removals of submandibular glands but not of adrenal glands completely prevented the postnephrectomy increases of plasma inactive renin. But, removals of submandibular glands or adrenal glands alone were followed by no changes in the basal levels of plasma inactive renin.(ABSTRACT TRUNCATED AT 400 WORDS)